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K Number
K180966
Device Name
FilmArray Pneumonia Panel
Manufacturer
BioFire Diagnostics, LLC
Date Cleared
2018-11-09

(210 days)

Product Code
QDPODPQBH
Regulation Number
866.3985
AI/MLSaMDIVD (In Vitro Diagnostic)TherapeuticDiagnosticis PCCP Authorized
Intended Use
The FilmArray® Pneumonia Panel is a multiplexed nucleic acid test intended for use with FilmArray® 2.0, or FilmArray® Torch systems for the simultaneous detection of multiple respiratory viral and bacterial nucleic acids, as well as select antimicrobial resistance genes, in sputum-like specimens (induced or expectorated sputum, or endotracheal aspirates) or bronchoalveolar lavage (BAL)-like specimens (BAL or mini-BAL) obtained from individuals suspected of lower respiratory tract infection. The following bacteria are reported semi-quantitatively with bins representing approximately 10°4 10°5, 10°6, or ≥10°7 genomic copies of bacterial nucleic acid per milliliter (copies/mL) of specimen, to aid in estimating relative abundance of nucleic acid from these common bacteria within a specimen: Bacteria reported with bins of 10^4 10^5, 10^6, or ≥10^7 copies/mL - Acinetobacter calcoaceticus-baumannii complex - Enterobacter cloacae complex - · Escherichia coli - Haemophilus influenzae - Klebsiella aerogenes - Klebsiella oxytoca - Klebsiella pneumoniae group - Moraxella catarrhalis - · Proteus spp. - Pseudomonas aeruginosa - Serratia marcescens - Staphylococcus aureus - Streptococcus agalactiae - Streptococcus pneumoniae - Streptococcus pyogenes The following atypical bacteria, viruses, and antimicrobial resistance genes are reported qualitatively: Atypical Bacteria - Chlamydia pneumoniae - Legionella pneumophila - Mycoplasma pneumoniae Viruses - Adenovirus - Coronavirus - Human Metapneumovirus - · Human Rhinovirus/Enterovirus - · Influenza A - · Influenza B - Parainfluenza Virus - Respiratory Syncytial Virus Antimicrobial Resistance Genes - CTX-M - IMP - КРС - NDM - · OXA-48-like - VIM - · mecA/C and MREJ The detection and identification of specific viral and bacterial nucleic acids, as well as the estimation of relative abundance of nucleic acid from common bacterial analytes, within specimens collected from individuals exhibiting signs and/or symptoms of a respiratory infection, aids in the diagnosis of lower respiratory infection if used in conjunction with other clinical and epidemiological information. The results of this test should not be used as for diagnosis, treatment, or other patient management decisions. Negative results in the setting of a respiratory illness may be due to infection with pathogens that are not detected by this test, pathogens below the limit of detection, or in the case of bacterial analytes, present at levels below the lowest reported 10°4 copies/mL bin. Detection of analytes does not rule out co-infection with other organisms: the agent(s) detected by the Film Array Pneumonia Panel may not be the definite cause of disease. Additional laboratory testing (e.g. bacterial and viral culture, immunofluorescence, and radiography) may be necessary when evaluating a patient with possible lower respiratory tract infection. Detection of bacterial nucleic acid may be indicative of colonizing or normal respiratory flora and may not indicate the causative agent of pneumonia. Semi-quantitative bin (copies/mL) results generated by the FilmArray Pneumonia Panel are not equivalent to CFU/mL and do not consistently correlate with the quantity of bacterial analytes compared to CFU/mL. For specimens with multiple bacteria detected, the relative abundance of nucleic acids (copies/mL) may not correlate with the relative abundance of bacteria as determined by culture (CFU/mL). Clinical correlation is advised to determine significance of semi-quantitative bin (copies/mL) for clinical management. The antimicrobial resistance gene detected may or may not be associated with the agent(s) responsible for disease. Negative results for these antimicrobial resistance gene assays do not indicate susceptibility to corresponding classes of antimicrobials, as multiple mechanisms of antimicrobial resistance exist. Antimicrobial resistance can occur via multiple mechanisms. A "Not Detected" result for a genetic marker of antimicrobial resistance does not indicate susceptibility to associated antimicrobial drugs or drug classes. A "Detected" result for a genetic marker of antimicrobial resistance cannot be definitively linked to the microorganism(s) detected. Culture is required to obtain isolates for antimicrobial susceptibility testing, and FilmArray Pneumonia Panel results should be used in conjunction with culture results for determination of bacterial susceptibility or resistance. Due to the genetic similarity between human rhinovirus and enterovirus, the test cannot reliably differentiate them. A positive Rhinovirus/Enterovirus result should be followed up using an alternate method (e.g., cell culture or sequence analysis) if differentiation is required. Culture is required to identify pathogens not detected by the FilmArray Pneumonia Panel, to further speciate analytes in genus, complex, or group results if desired, to identify bacterial pathogens present below the 10°4 copies/mL bin if desired, and for antimicrobial susceptibility testing.
Device Description
The FilmArray Pneumonia Panel is designed to simultaneously identify 26 potential pathogens of lower respiratory tract infection (LRTI) and associated antimicrobial resistance (AMR) genes from a sputumlike (induced and expectorated sputum as well as endotracheal aspirate, ETA) or bronchoalveolar lavage (BAL)-like (BAL and mini-BAL) specimens obtained from individuals with signs and/or symptoms of lower respiratory tract infection in a time (~1 hour) that allows the test results to be used in determining appropriate patient treatment and management. FilmArray Pneumonia Panel is compatible with BioFire Diagnostics' (BioFire) PCR-based in vitro diagnostic FilmArray 2.0, and FilmArray Torch systems for infectious disease testing. A specific software module (i.e. FilmArray Pneumonia Panel pouch module) is used to perform FilmArray Pneumonia Panel testing on these systems. Bacteria - Quantitative Results | Antimicrobial Resistance Genes -----------------------------------------------|---------------------------------------------------- Acinetobacter calcoaceticus-baumannii complex | blaCTX-M (Extended spectrum beta-lactamase (ESBL)) Enterobacter cloacae complex | blaIMP (Carbapenem resistance) Escherichia coli | blaKPC (Carbapenem resistance) Haemophilus influenzae | mecA/mecC and MREJ (Methicillin resistance) Klebsiella aerogenes | blaNDM (Carbapenem resistance) Klebsiella oxytoca | blaOXA48-like (Carbapenem resistance) Klebsiella pneumoniae group | blaVIM (Carbapenem resistance) Moraxella catarrhalis | Proteus spp. | Adenovirus Pseudomonas aeruginosa | Coronavirus Serratia marcescens | Human Metapneumovirus Staphylococcus aureus | Human Rhinovirus/Enterovirus Streptococcus agalactiae | Influenza A Streptococcus pneumoniae | Influenza B Streptococcus pyogenes | Bacteria (Atypical) - Qualitative Results | Viruses Chlamydia pneumoniae | Parainfluenza Virus Legionella pneumophila | Respiratory Syncytial Virus Mycoplasma pneumoniae | A test is initiated by loading Hydration Solution into one port of the FilmArray pouch and a sputum-like or BAL-like sample mixed with the provided Sample Buffer into the other port of the FilmArray Pneumonia Panel pouch and placing it in a FilmArray instrument. The pouch contains all of the reagents required for specimen testing and analysis in a freeze-dried format; the addition of Hydration Solution and Sample/Buffer Mix rehydrates the reagents. After the pouch is prepared, the FilmArray Software guides the user though the steps of placing the instrument, scanning the pouch barcode, entering the sample identification, and initiating the run. The FilmArray instrument contains a coordinated system of inflatable bladders and seal points, which act on the pouch to control the movement of liquid between the pouch blisters. When a bladder is inflated over a reagent blister, it forces liquid from the blister into connecting channels. Alternatively, when a seal is placed over a connecting channel it acts as a valve to open or close a channel. In addition, electronically-controlled pneumatic pistons are positioned over multiple plungers in order to deliver the rehydrated reagents into the blisters at the appropriate times. Two Peltier devices control heating and cooling of the pouch to drive the PCR reactions and the melt curve analysis. Nucleic acid extraction occurs within the FilmArray pouch using mechanical and chemical lysis followed by purification using standard magnetic bead technology. After extracting and purifying nucleic acids from the unprocessed sample, the FilmArray performs a nested multiplex PCR that is executed in two stages. During the filmArray performs a single, large volume, highly multiplexed reverse transcription PCR (rt-PCR) reaction. The products from first stage PCR are then diluted and combined with a fresh, primer-free master mix and a fluorescent double stranded DNA binding dye (LC Green® Plus, BioFire Diagnostics). The solution is then distributed to each well of the array. Array wells contain sets of primers designed specifically to amplify sequences internal to the PCR products generated during the first stage PCR reaction. The 2nd stage PCR, or nested PCR, is performed in singleplex fashion in each well of the array. At the conclusion of the 2nd stage PCR, the array is interrogated by melt curve analysis for the detection of signature amplicons denoting the presence of specific targets. A digital camera placed in front of the 2nd stage PCR captures fluorescent images of the PCR reactions and software interprets the data. The FilmArray Software automatically interprets the results of each DNA melt curve analysis and combines the data with the results of the internal pouch controls to provide a test result for each organism on the panel.
More Information

DEN170047

Not Found

No
The description focuses on the molecular biology techniques (PCR, melt curve analysis) and the automated fluid handling system within the device to detect and identify specific nucleic acid targets. There is no mention of AI or ML being used for data interpretation, pattern recognition, or decision support beyond the automated interpretation of melt curve analysis based on predefined parameters.

No.
Explanation: The device is a diagnostic test (a multiplexed nucleic acid test) used to detect pathogens and antimicrobial resistance genes. It is explicitly stated that "The results of this test should not be used as for diagnosis, treatment, or other patient management decisions." This indicates it provides information to aid in diagnosis but does not directly treat or manage a condition, which is the definition of a therapeutic device.

Yes

The text states that the device "aids in the diagnosis of lower respiratory infection if used in conjunction with other clinical and epidemiological information." This clearly indicates its role in assisting the diagnostic process.

No

The device description explicitly states that the FilmArray Pneumonia Panel is used with the FilmArray 2.0 or FilmArray Torch systems, which are hardware instruments. The process involves loading a physical pouch containing reagents into the instrument, and the instrument performs physical actions (bladder inflation, seals, pistons, Peltier devices) to process the sample and perform PCR. While software is used to guide the user, interpret data, and provide results, the core functionality relies on dedicated hardware and physical reagents.

Based on the provided information, the FilmArray® Pneumonia Panel is indeed an IVD (In Vitro Diagnostic).

Here's why:

  • Intended Use: The document explicitly states the device is "intended for use with FilmArray® 2.0, or FilmArray® Torch systems for the simultaneous detection of multiple respiratory viral and bacterial nucleic acids, as well as select antimicrobial resistance genes, in sputum-like specimens... or bronchoalveolar lavage (BAL)-like specimens... obtained from individuals suspected of lower respiratory tract infection." This describes a test performed in vitro (outside the body) on biological specimens to provide information about a patient's health status.
  • Purpose: The intended use further clarifies that the detection and identification of these nucleic acids "aids in the diagnosis of lower respiratory infection if used in conjunction with other clinical and epidemiological information." This directly aligns with the definition of an IVD, which is used to diagnose or aid in the diagnosis of a disease or condition.
  • Device Description: The description details a system that processes biological samples (sputum-like and BAL-like specimens) using laboratory techniques (nucleic acid extraction, PCR, melt curve analysis) to generate results. This is characteristic of an IVD device.
  • Regulatory Context: The mention of a "Predicate Device(s)" with a K number (DEN170047) strongly indicates that this device is undergoing or has undergone regulatory review as a medical device, specifically an IVD, by a regulatory body like the FDA in the US. K numbers are associated with 510(k) submissions for medical devices, including IVDs.
  • Performance Studies: The detailed descriptions of clinical performance studies using patient specimens and comparisons to reference methods are standard requirements for demonstrating the analytical and clinical validity of an IVD.

Therefore, the FilmArray® Pneumonia Panel fits the definition and characteristics of an In Vitro Diagnostic device.

N/A

Intended Use / Indications for Use

The FilmArray® Pneumonia Panel is a multiplexed nucleic acid test intended for use with FilmArray® 2.0, or FilmArray® Torch systems for the simultaneous detection of multiple respiratory viral and bacterial nucleic acids, as well as select antimicrobial resistance genes, in sputum-like specimens (induced or expectorated sputum, or endotracheal aspirates) or bronchoalveolar lavage (BAL)-like specimens (BAL or mini-BAL) obtained from individuals suspected of lower respiratory tract infection.
The following bacteria are reported semi-quantitatively with bins representing approximately 10^4 10^5, 10^6, or ≥10^7 genomic copies of bacterial nucleic acid per milliliter (copies/mL) of specimen, to aid in estimating relative abundance of nucleic acid from these common bacteria within a specimen:
Bacteria reported with bins of 10^4 10^5, 10^6, or ≥10^7 copies/mL

  • Acinetobacter calcoaceticus-baumannii complex
  • Enterobacter cloacae complex
  • · Escherichia coli
  • Haemophilus influenzae
  • Klebsiella aerogenes
  • Klebsiella oxytoca
  • Klebsiella pneumoniae group
  • Moraxella catarrhalis
  • · Proteus spp.
  • Pseudomonas aeruginosa
  • Serratia marcescens
  • Staphylococcus aureus
  • Streptococcus agalactiae
  • Streptococcus pneumoniae
  • Streptococcus pyogenes

The following atypical bacteria, viruses, and antimicrobial resistance genes are reported qualitatively:
Atypical Bacteria

  • Chlamydia pneumoniae
  • Legionella pneumophila
  • Mycoplasma pneumoniae
    Viruses
  • Adenovirus
  • Coronavirus
  • Human Metapneumovirus
  • · Human Rhinovirus/Enterovirus
  • · Influenza A
  • · Influenza B
  • Parainfluenza Virus
  • Respiratory Syncytial Virus
    Antimicrobial Resistance Genes
  • CTX-M
  • IMP
  • КРС
  • NDM
  • · OXA-48-like
  • VIM
  • · mecA/C and MREJ

The detection and identification of specific viral and bacterial nucleic acids, as well as the estimation of relative abundance of nucleic acid from common bacterial analytes, within specimens collected from individuals exhibiting signs and/or symptoms of a respiratory infection, aids in the diagnosis of lower respiratory infection if used in conjunction with other clinical and epidemiological information. The results of this test should not be used as for diagnosis, treatment, or other patient management decisions.
Negative results in the setting of a respiratory illness may be due to infection with pathogens that are not detected by this test, pathogens below the limit of detection, or in the case of bacterial analytes, present at levels below the lowest reported 10^4 copies/mL bin. Detection of analytes does not rule out co-infection with other organisms: the agent(s) detected by the Film Array Pneumonia Panel may not be the definite cause of disease. Additional laboratory testing (e.g. bacterial and viral culture, immunofluorescence, and radiography) may be necessary when evaluating a patient with possible lower respiratory tract infection.
Detection of bacterial nucleic acid may be indicative of colonizing or normal respiratory flora and may not indicate the causative agent of pneumonia. Semi-quantitative bin (copies/mL) results generated by the FilmArray Pneumonia Panel are not equivalent to CFU/mL and do not consistently correlate with the quantity of bacterial analytes compared to CFU/mL. For specimens with multiple bacteria detected, the relative abundance of nucleic acids (copies/mL) may not correlate with the relative abundance of bacteria as determined by culture (CFU/mL). Clinical correlation is advised to determine significance of semi-quantitative bin (copies/mL) for clinical management.
The antimicrobial resistance gene detected may or may not be associated with the agent(s) responsible for disease. Negative results for these antimicrobial resistance gene assays do not indicate susceptibility to corresponding classes of antimicrobials, as multiple mechanisms of antimicrobial resistance exist.
Antimicrobial resistance can occur via multiple mechanisms. A "Not Detected" result for a genetic marker of antimicrobial resistance does not indicate susceptibility to associated antimicrobial drugs or drug classes. A "Detected" result for a genetic marker of antimicrobial resistance cannot be definitively linked to the microorganism(s) detected. Culture is required to obtain isolates for antimicrobial susceptibility testing, and FilmArray Pneumonia Panel results should be used in conjunction with culture results for determination of bacterial susceptibility or resistance.
Due to the genetic similarity between human rhinovirus and enterovirus, the test cannot reliably differentiate them. A positive Rhinovirus/Enterovirus result should be followed up using an alternate method (e.g., cell culture or sequence analysis) if differentiation is required.
Culture is required to identify pathogens not detected by the FilmArray Pneumonia Panel, to further speciate analytes in genus, complex, or group results if desired, to identify bacterial pathogens present below the 10^4 copies/mL bin if desired, and for antimicrobial susceptibility testing.

Product codes (comma separated list FDA assigned to the subject device)

ODP, QBH

Device Description

The FilmArray Pneumonia Panel is designed to simultaneously identify 26 potential pathogens of lower respiratory tract infection (LRTI) and associated antimicrobial resistance (AMR) genes from a sputumlike (induced and expectorated sputum as well as endotracheal aspirate, ETA) or bronchoalveolar lavage (BAL)-like (BAL and mini-BAL) specimens obtained from individuals with signs and/or symptoms of lower respiratory tract infection in a time (~1 hour) that allows the test results to be used in determining appropriate patient treatment and management. FilmArray Pneumonia Panel is compatible with BioFire Diagnostics' (BioFire) PCR-based in vitro diagnostic FilmArray 2.0, and FilmArray Torch systems for infectious disease testing. A specific software module (i.e. FilmArray Pneumonia Panel pouch module) is used to perform FilmArray Pneumonia Panel testing on these systems.
Bacteria - Quantitative Results: Acinetobacter calcoaceticus-baumannii complex, Enterobacter cloacae complex, Escherichia coli, Haemophilus influenzae, Klebsiella aerogenes, Klebsiella oxytoca, Klebsiella pneumoniae group, Moraxella catarrhalis, Proteus spp., Pseudomonas aeruginosa, Serratia marcescens, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus pneumoniae, Streptococcus pyogenes
Antimicrobial Resistance Genes: blaCTX-M (Extended spectrum beta-lactamase (ESBL)), blaIMP (Carbapenem resistance), blaKPC (Carbapenem resistance), mecA/mecC and MREJ (Methicillin resistance), blaNDM (Carbapenem resistance), blaOXA48-like (Carbapenem resistance), blaVIM (Carbapenem resistance)
Bacteria (Atypical) - Qualitative Results: Chlamydia pneumoniae, Legionella pneumophila, Mycoplasma pneumoniae
Viruses: Adenovirus, Coronavirus, Human Metapneumovirus, Human Rhinovirus/Enterovirus, Influenza A, Influenza B, Parainfluenza Virus, Respiratory Syncytial Virus
A test is initiated by loading Hydration Solution into one port of the FilmArray pouch and a sputum-like or BAL-like sample mixed with the provided Sample Buffer into the other port of the FilmArray Pneumonia Panel pouch and placing it in a FilmArray instrument. The pouch contains all of the reagents required for specimen testing and analysis in a freeze-dried format; the addition of Hydration Solution and Sample/Buffer Mix rehydrates the reagents. After the pouch is prepared, the FilmArray Software guides the user though the steps of placing the instrument, scanning the pouch barcode, entering the sample identification, and initiating the run.
The FilmArray instrument contains a coordinated system of inflatable bladders and seal points, which act on the pouch to control the movement of liquid between the pouch blisters. When a bladder is inflated over a reagent blister, it forces liquid from the blister into connecting channels. Alternatively, when a seal is placed over a connecting channel it acts as a valve to open or close a channel. In addition, electronically-controlled pneumatic pistons are positioned over multiple plungers in order to deliver the rehydrated reagents into the blisters at the appropriate times. Two Peltier devices control heating and cooling of the pouch to drive the PCR reactions and the melt curve analysis.
Nucleic acid extraction occurs within the FilmArray pouch using mechanical and chemical lysis followed by purification using standard magnetic bead technology. After extracting and purifying nucleic acids from the unprocessed sample, the FilmArray performs a nested multiplex PCR that is executed in two stages. During the filmArray performs a single, large volume, highly multiplexed reverse transcription PCR (rt-PCR) reaction. The products from first stage PCR are then diluted and combined with a fresh, primer-free master mix and a fluorescent double stranded DNA binding dye (LC Green® Plus, BioFire Diagnostics). The solution is then distributed to each well of the array. Array wells contain sets of primers designed specifically to amplify sequences internal to the PCR products generated during the first stage PCR reaction. The 2nd stage PCR, or nested PCR, is performed in singleplex fashion in each well of the array. At the conclusion of the 2nd stage PCR, the array is interrogated by melt curve analysis for the detection of signature amplicons denoting the presence of specific targets. A digital camera placed in front of the 2nd stage PCR captures fluorescent images of the PCR reactions and software interprets the data.
The FilmArray Software automatically interprets the results of each DNA melt curve analysis and combines the data with the results of the internal pouch controls to provide a test result for each organism on the panel.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

lower respiratory tract, respiratory tract

Indicated Patient Age Range

all ages

Intended User / Care Setting

Not Found

Description of the training set, sample size, data source, and annotation protocol

Not Found

Description of the test set, sample size, data source, and annotation protocol

The clinical performance of the FilmArray Pneumonia Panel was established during a multi-center study conducted at eight geographically distinct U.S. study sites from October 2016 to July 2017. A total of 904 residual BAL (821 BAL and 83 mini-BAL) and 925 residual sputum ( 478 sputum and 447 ETA) specimens were acquired for the prospective clinical study. FilmArray Pneumonia Panel performance in BAL and mini-BAL was similar, as was performance in sputum and ETA; therefore these sample types are not stratified further in performance tables. A total of 58 BAL and 89 sputum specimens were excluded from the final data analysis. The most common reasons for specimen exclusion for both specimen types was reference culture unable to be performed, the specimen was found to not meet the inclusion criteria after the specimen had been enrolled, or the study site was unable to complete the Case Report Form (CRF). The final data set consisted of 846 BAL and 836 sputum specimens.
All specimens were evaluated with the FilmArray Pneumonia Panel at clinical study sites. Refrigerated specimen aliquots were sent to a central reference laboratory for quantitative reference culture (qRefCx) and frozen specimen aliquots were also sent to BioFire for evaluation by polymerase chain reaction (PCR)/sequencing-based comparator methods.
The reference methods used in this study were as follows:
Bacterial analytes were compared to qRefCx to evaluate sensitivity and the method was considered positive for the presence of the organism of interest if it was recovered in culture and enumerated at a level of 3162 (10^3.5) CFU/mL or greater.
Bacterial analytes were also evaluated by comparison to a single PCR assay for the organism of interest followed by a quantitative molecular assay that included sequencing (qMol) to assess FilmArray bin reporting performance. Atypical bacteria and viruses were compared to two conventional PCR assays followed by bidirectional sequencing. For specimens with an applicable bacteria detected by FilmArray, AMR genes were compared to a single PCR assay (from the specimen) followed by sequencing. A specimen was considered to be positive for an analyte if bi-directional sequencing data meeting predefined quality acceptance criteria matched organism-specific sequences deposited in the NCBI GenBank database (www.ncbi.nlm.nih.gov) with acceptable E-values. When two PCR comparator assays were used, any specimen that tested negative by both of the comparator assays was considered Negative.
Positive Percent Agreement (PPA) or Sensitivity for each analyte was calculated as 100% x (TP + FN)). True positive (TP) indicates that both the FilmArray Pneumonia Panel and the comparator method had a positive result for this specific analyte, and false negative (FN) indicates that the FilmArray Pneumonia Panel result was negative while the comparator result was positive. Negative Percent Agreement (NPA) or Specificity was calculated as 100% x (TN + FP)). True negative (TN) indicates that both the FilmArray Pneumonia Panel and the comparator method had negative results, and a false positive (FP) indicates that the FilmArray Pneumonia Panel result was positive but the comparator result was negative. The exact binomial two-sided 95% confidence interval was calculated. Samples for which false positive and/or false negative results (i.e., discrepant results) were obtained when comparing the FilmArray Pneumonia Panel results to the comparator method results were further investigated. The discrepancy investigations were primarily performed as follows: for discrepancies between the Pneumonia Panel and reference culture for bacterial analytes, discrepancies were first examined to see if qRefCx or FilmArray had observed the analyte but reported it as "negative" or "Not Detected" because it was below the detection threshold. If this did not resolve the discrepancy, the results of qMol testing were considered. And if these methods still did not resolve the discrepancy, it was then investigated in the same manner as other analytes that used molecular comparator (i.e. using multiple additional molecular assays followed by sequence analysis). The results of SOC testing were also considered.

Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)

Clinical Performance: Multi-center study conducted at eight geographically distinct U.S. study sites from October 2016 to July 2017. A total of 904 residual BAL (821 BAL and 83 mini-BAL) and 925 residual sputum (478 sputum and 447 ETA) specimens were acquired. The final data set consisted of 846 BAL and 836 sputum specimens.
Key Results for BAL specimens:
Acinetobacter calcoaceticus-baumannii complex: Sensitivity/PPA 0/0 (-), Specificity/NPA 99.2% (98.3-99.6%)
Klebsiella aerogenes: Sensitivity/PPA 85.7% (48.7-97.4%), Specificity/NPA 99.2% (98.3-99.6%)
Enterobacter cloacae complex: Sensitivity/PPA 91.7% (64.6-98.5%), Specificity/NPA 98.6% (97.5-99.2%)
Escherichia coli: Sensitivity/PPA 100% (75.8-100%), Specificity/NPA 99.0% (98.1-99.5%)
Haemophilus influenzae: Sensitivity/PPA 100% (72.2-100%), Specificity/NPA 91.4% (89.3-93.1%)
Klebsiella oxytoca: Sensitivity/PPA 100% (34.2-100%), Specificity/NPA 98.9% (98.0-99.4%)
Many other results are provided.

Testing of Preselected Archived Specimens: Evaluation of 171 frozen archived specimens (18 negatives, 153 positives) at BioFire. 149 positive specimens (173 analytes) were used for analysis.
Key Results:
BAL Specimens:
Acinetobacter calcoaceticus-baumannii complex: PPA 100% (51.0-100%), NPA 100% (93.2-100%)
Klebsiella aerogenes: PPA 50.0%, NPA 100% (93.2-100%)
Proteus spp.: PPA 80.0% (37.6-96.4%), NPA 100% (92.6-100%)
Serratia marcescens: PPA 100% (72.2-100%), NPA 100% (92.3-100%)
Streptococcus pyogenes: PPA 100%, NPA 100% (93.7-100%)
CTX-M: PPA 100% (64.6-100%), NPA 100% (88.3-100%)
Chlamydia pneumoniae: PPA 100%, NPA 100% (95.9-100%)
Legionella pneumophila: PPA 100%, NPA 100% (93.7-100%)
Adenovirus: PPA 100% (67.6-100%), NPA 100% (95.5-100%)
Human Metapneumovirus: PPA 100% (74.1-100%), NPA 100% (95.2-100%)
Influenza A: PPA 100% (84.5-100%), NPA 100% (94.7-100%)
Influenza B: PPA 100% (43.9-100%), NPA 100% (95.7-100%)
Parainfluenza Virus: PPA 100% (81.6-100%), NPA 99.0% (92.2-99.7%)
Respiratory Syncytial Virus: PPA 93.8% (71.7-98.9%), NPA 100% (95.1-100%)

Sputum Specimens:
Streptococcus pyogenes: PPA 100% (64.6-100%), NPA 100% (67.6-100%)
CTX-M: PPA 100%, NPA 100% (75.8-100%)
Influenza A: PPA 100% (34.2-100%), NPA 0%

Testing of Contrived Specimens: 1125 contrived specimens were spiked using residual clinical samples.
Key Results for BAL Specimens:
Acinetobacter calcoaceticus-baumannii complex: Sensitivity/PPA 94.0% (83.8-97.9%), Specificity/NPA 100% (99.4-100%)
Klebsiella aerogenes: Sensitivity/PPA 85.5% (73.8-92.4%), Specificity/NPA 99.7% (98.8-99.9%)
Klebsiella oxytoca: Sensitivity/PPA 92.0% (81.2-96.8%), Specificity/NPA 100% (99.4-100%)
Proteus spp.: Sensitivity/PPA 96.0% (86.5-98.9%), Specificity/NPA 100% (99.4-100%)

Testing of Polymicrobial Contrived Specimens: Two sets of individual BAL (N=60) and sputum (N=60) specimens were multi-spiked.
Key Results: Majority of spiked organisms were reported at expected bin level. One false negative for P. mirabilis in BAL.

Selected Analytical Studies - Limit of Detection: LoD established for atypical bacteria and viruses.
Key Results: For C. pneumoniae LoD is 5.0E-01 TCID50/mL and 3.3E+02 copies/mL. For L. pneumophila LoD is 5.0E+02 CFU/mL and 1.6E+03 copies/mL. For M. pneumoniae LoD is 7.5E+01 TCID50/mL and 3.5E+03 copies/mL. Viruses also show similar dual LoD values.

Inclusivity: Analytical reactivity evaluated via empirical testing (>350 genetically diverse organisms) and in silico analysis.
Key Results: Atypical bacteria and viruses detected within 3xLoD. 94.4% of bacterial isolates detected with expected bin results at 1.0E+04 copies/mL.

Analytical Specificity (Cross-Reactivity and Exclusivity): Evaluated by in silico analyses and empirical testing of high concentrations.
Key Results:
Closely-Related Species: Escherichia fergusonii, Shigella species, Klebsiella michiganensis, Staphylococcus argenteus, Staphylococcus schweitzeri, Pseudomonas putida.
Unrelated Species: Bordetella species, Aspergillus niger, Cryptococcus laurentii, Cryptococcus uniguttulatus, Stenotrophomonas acidaminiphila, Acinetobacter schindleri, Burkholderia vietnamiensis, Lelliottia amnigena, Enterobacter kobei, Enterobacter ludwigii, Enterobacter cloacae.

Precision (Reproducibility): Testing performed over multiple days at three laboratory locations using FilmArray 2.0 and FilmArray Torch systems (30 tests/system, 90 total replicates/sample/concentration).
Key Results: High agreement (98.9% to 100%) with expected detected/not detected results for atypical bacteria and viruses. Bacterial bin precision varied based on concentration, adhering to a probabilistic model, achieving >90% at bin center and ~50% at bin limits. AMR gene detection reproducibility also high (99.2% to 100% for reportable range).

Interference: Potentially interfering substances evaluated.
Key Results: Bleach, MycoPrep, 2% NaOH, and 5% Oxalic acid interfered by damaging nucleic acids. All other substances tested (endogenous, exogenous, competitive microorganisms, other disinfectants) showed no interference.

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

Positive Percent Agreement (PPA) or Sensitivity: Calculated as 100% x (TP / (TP + FN)).
Negative Percent Agreement (NPA) or Specificity: Calculated as 100% x (TN / (TN + FP)).
Exact binomial two-sided 95% confidence interval was calculated for PPA and NPA.

Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.

DEN170047

Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).

Not Found

§ 866.3985 Device to detect and identify microorganisms and associated resistance marker nucleic acids directly in respiratory specimens.

(a)
Identification. A device to detect and identify microorganisms and associated resistance marker nucleic acids directly from respiratory specimens is an in vitro diagnostic device intended for the detection and identification of microorganisms and associated resistance markers in respiratory specimens collected from patients with signs or symptoms of respiratory infection. The device is intended to aid in the diagnosis of respiratory infection in conjunction with clinical signs and symptoms and other laboratory findings. These devices do not provide confirmation of antibiotic susceptibility since mechanisms of resistance may exist other than those detected by the device.(b)
Classification. Class II (special controls). The special controls for this device are:(1) The intended use for the 21 CFR 809.10 labeling must include a detailed description of what the device detects, the type of results provided to the user, the clinical indications appropriate for test use, and the specific population(s) for which the device is intended.
(2) The 21 CFR 809.10(b) labeling must include:
(i) A detailed device description, including all device components, control elements incorporated into the test procedure, instrument requirements, ancillary reagents required but not provided, and a detailed explanation of the methodology, including all pre-analytical methods for processing of specimens.
(ii) Performance characteristics from analytical studies, including, but not limited to, limit of detection, inclusivity, reproducibility, cross reactivity, interfering substances, competitive inhibition, carryover/cross contamination, specimen stability, and linearity, as applicable.
(iii) A limiting statement that the device is intended to be used in conjunction with clinical history, signs and symptoms, and results of other diagnostic tests, including culture and antimicrobial susceptibility testing.
(iv) A detailed explanation of the interpretation of test results for clinical specimens and acceptance criteria for any quality control testing.
(v) A limiting statement that negative results for microorganisms do not preclude the possibility of infection, and should not be used as the sole basis for diagnosis, treatment, or other patient management decisions.
(vi) If applicable, a limiting statement that detected microorganisms may not be the cause of lower respiratory tract infection and may be indicative of colonizing or normal respiratory flora.
(vii) If applicable, a limiting statement that detection of resistance markers cannot be definitively linked to specific microorganisms and that the source of a detected resistance marker may be an organism not detected by the assay, including colonizing flora.
(viii) If applicable, a limiting statement that detection of antibiotic resistance markers may not correlate with phenotypic gene expression.
(3) The 21 CFR 809.10(b) labeling and any test report generated by the device must include a limiting statement that negative results for resistance markers do not indicate susceptibility of detected microorganisms.
(4) Design verification and validation must include:
(i) Performance characteristics from clinical studies that include prospective (sequential) samples and, if appropriate, additional characterized samples. The study must be performed on a study population consistent with the intended use population and compare the device performance to results obtained from an FDA accepted reference method and/or FDA accepted comparator method, as appropriate. Results from the clinical studies must include the clinical study protocol (including predefined statistical analysis plan, if applicable), clinical study report, and results of all statistical analyses.
(ii) A detailed device description including the following:
(A) Thorough description of the assay methodology including, but not limited to, primer/probe sequences, primer/probe design, and rationale for target sequence selection, as applicable.
(B) Algorithm used to generate a final result from raw data (e.g., how raw signals are converted into a reported result).
(iii) A detailed description of device software, including, but not limited to, validation activities and outcomes.
(iv) As part of the risk management activities, an appropriate end user device training program must be offered as an effort to mitigate the risk of failure from user error.

0

Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

BioFire Diagnostics, LLC Kristen Kanack Senior Vice President, Regulatory & Clinical Affairs 515 Colorow Drive Salt Lake City, Utah 84108

Re: K180966

Trade/Device Name: FilmArray Pneumonia Panel Regulation Number: 21 CFR 866.3985 Regulation Name: Device to detect and identify microorganisms and associated resistance marker nucleic acids directly in respiratory specimens Regulatory Class: Class II Product Code: ODP

Dear Kristen Kanack:

The Food and Drug Administration (FDA) is sending this letter to notify you of an administrative change related to your previous substantial equivalence (SE) determination letter dated November 9th, 2018. Specifically, FDA is updating this SE Letter as an administrative correction. because FDA has created a new product code to better categorize your device technology.

Please note that the 510(k) submission was not re-reviewed. For questions regarding this letter please contact Kristian Roth, Ph.D., OHT7: Office of In Vitro Diagnostics and Radiological Health,

Sincerely,

Digitally signed by Kristian M. Roth Kristian M. Roth -S Date: 2021.06.24 08:49:06 -04'00

Kristian Roth, Ph.D. Deputy Director (Acting) Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health

1

Image /page/1/Picture/0 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, with the letters "FDA" in a blue square, followed by the words "U.S. FOOD & DRUG" in blue, and the word "ADMINISTRATION" in a smaller font below.

November 9, 2018

BioFire Diagnostics, LLC Kristen Kanack Senior Vice President, Regulatory & Clinical Affairs 515 Colorow Drive Salt Lake City, Utah 84108

Re: K180966

Trade/Device Name: FilmArray Pneumonia Panel Regulation Number: 21 CFR 866.3985 Regulation Name: Device to detect and identify microorganisms and associated resistance marker nucleic acids directly in respiratory specimens Regulatory Class: Class II Product Code: QBH Dated: April 12, 2018 Received: April 13, 2018

Dear Kristen Kanack:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpm/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's

2

requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.htm ); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm.

For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely,

Steven R. Gitterman -S for

Uwe Scherf, Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

3

Indications for Use

510(k) Number (if known) K180966

Device Name FilmArray Pneumonia Panel

Indications for Use (Describe)

The FilmArray® Pneumonia Panel is a multiplexed nucleic acid test intended for use with FilmArray® 2.0, or FilmArray® Torch systems for the simultaneous detection of multiple respiratory viral and bacterial nucleic acids, as well as select antimicrobial resistance genes, in sputum-like specimens (induced or expectorated sputum, or endotracheal aspirates) or bronchoalveolar lavage (BAL)-like specimens (BAL or mini-BAL) obtained from individuals suspected of lower respiratory tract infection.

The following bacteria are reported semi-quantitatively with bins representing approximately 10°4 10°5, 10°6, or ≥10°7 genomic copies of bacterial nucleic acid per milliliter (copies/mL) of specimen, to aid in estimating relative abundance of nucleic acid from these common bacteria within a specimen:

Bacteria reported with bins of 10^4 10^5, 10^6, or ≥10^7 copies/mL

  • Acinetobacter calcoaceticus-baumannii complex
  • Enterobacter cloacae complex
  • · Escherichia coli
  • Haemophilus influenzae
  • Klebsiella aerogenes
  • Klebsiella oxytoca
  • Klebsiella pneumoniae group
  • Moraxella catarrhalis
  • · Proteus spp.
  • Pseudomonas aeruginosa
  • Serratia marcescens
  • Staphylococcus aureus
  • Streptococcus agalactiae
  • Streptococcus pneumoniae
  • Streptococcus pyogenes

The following atypical bacteria, viruses, and antimicrobial resistance genes are reported qualitatively:

Atypical Bacteria

  • Chlamydia pneumoniae
  • Legionella pneumophila
  • Mycoplasma pneumoniae

Viruses

  • Adenovirus
  • Coronavirus
  • Human Metapneumovirus
  • · Human Rhinovirus/Enterovirus
  • · Influenza A
  • · Influenza B
  • Parainfluenza Virus
  • Respiratory Syncytial Virus

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Antimicrobial Resistance Genes

  • CTX-M
  • IMP
  • КРС
  • NDM
  • · OXA-48-like
  • VIM
  • · mecA/C and MREJ

The detection and identification of specific viral and bacterial nucleic acids, as well as the estimation of relative abundance of nucleic acid from common bacterial analytes, within specimens collected from individuals exhibiting signs and/or symptoms of a respiratory infection, aids in the diagnosis of lower respiratory infection if used in conjunction with other clinical and epidemiological information. The results of this test should not be used as for diagnosis, treatment, or other patient management decisions.

Negative results in the setting of a respiratory illness may be due to infection with pathogens that are not detected by this test, pathogens below the limit of detection, or in the case of bacterial analytes, present at levels below the lowest reported 10°4 copies/mL bin. Detection of analytes does not rule out co-infection with other organisms: the agent(s) detected by the Film Array Pneumonia Panel may not be the definite cause of disease. Additional laboratory testing (e.g. bacterial and viral culture, immunofluorescence, and radiography) may be necessary when evaluating a patient with possible lower respiratory tract infection.

Detection of bacterial nucleic acid may be indicative of colonizing or normal respiratory flora and may not indicate the causative agent of pneumonia. Semi-quantitative bin (copies/mL) results generated by the FilmArray Pneumonia Panel are not equivalent to CFU/mL and do not consistently correlate with the quantity of bacterial analytes compared to CFU/mL. For specimens with multiple bacteria detected, the relative abundance of nucleic acids (copies/mL) may not correlate with the relative abundance of bacteria as determined by culture (CFU/mL). Clinical correlation is advised to determine significance of semi-quantitative bin (copies/mL) for clinical management.

The antimicrobial resistance gene detected may or may not be associated with the agent(s) responsible for disease. Negative results for these antimicrobial resistance gene assays do not indicate susceptibility to corresponding classes of antimicrobials, as multiple mechanisms of antimicrobial resistance exist.

Antimicrobial resistance can occur via multiple mechanisms. A "Not Detected" result for a genetic marker of antimicrobial resistance does not indicate susceptibility to associated antimicrobial drugs or drug classes. A "Detected" result for a genetic marker of antimicrobial resistance cannot be definitively linked to the microorganism(s) detected. Culture is required to obtain isolates for antimicrobial susceptibility testing, and FilmArray Pneumonia Panel results should be used in conjunction with culture results for determination of bacterial susceptibility or resistance.

Due to the genetic similarity between human rhinovirus and enterovirus, the test cannot reliably differentiate them. A positive Rhinovirus/Enterovirus result should be followed up using an alternate method (e.g., cell culture or sequence analysis) if differentiation is required.

Culture is required to identify pathogens not detected by the FilmArray Pneumonia Panel, to further speciate analytes in genus, complex, or group results if desired, to identify bacterial pathogens present below the 10°4 copies/mL bin if desired, and for antimicrobial susceptibility testing.

Type of Use (Select one or both, as applicable)
-------------------------------------------------

| Over-The-Counter Use (21 CFR 801 Subpart C)

| Prescription Use (Part 21 CFR 801 Subpart D)

CONTINUE ON A SEPARATE PAGE IF NEEDED.

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510(k) Summary BioFire Diagnostics, LLC

FilmArray® Pneumonia Panel

Introduction:

According to the requirements of 21 CFR 807.92, the following information provides sufficient detail to understand the basis for a determination of substantial equivalence.

Submitted by:

BioFire Diagnostics, LLC 515 Colorow Drive Salt Lake City, UT 84108

Telephone: 801-736-6354 Facsimile: 801-588-0507

Contact: Kristen J. Kanack, ext. 1330

Date Submitted: April 12, 2018

Device Name and Classification:

Trade Name: FilmArray Pneumonia Panel

Regulation Number: 21 CFR 866.3985

Classification Name: Device to detect and identify microorganisms and associated resistance marker nucleic acids directly in respiratory specimens

Product Code: QDP

Predicate Device:

DEN170047 - Curetis Unyvero LRT Application

Intended Use:

The FilmArray® Pneumonia Panel is a multiplexed nucleic acid test intended for use with FilmArray® FilmArray® 2.0, or FilmArray® Torch systems for the simultaneous detection and identification of multiple respiratory viral and bacterial nucleic acids, as well as select antimicrobial resistance genes, in sputum-like specimens (induced or expectorated sputum, or endotracheal aspirates) or bronchoalveolar lavage (BAL)-like specimens (BAL or mini-BAL) obtained from individuals suspected of lower respiratory tract infection.

The following bacteria are reported semi-qualitatively with bins representing approximately 10^4, 10^5. 10°6. or >10°7 genomic copies of bacterial nucleic acid per milliliter (copies/mL) of specimen, to aid in estimating relative abundance of nucleic acid from these common bacteria within a specimen:

7

Bacteria reported with bins of 10 $^4$ , 10 $^5$ , 10 $^6$ , or ≥10 $^7$ copies/mL
Acinetobacter calcoaceticus-baumannii
complexKlebsiella oxytocaSerratia marcescens
Enterobacter cloacae complexKlebsiella pneumoniae groupStaphylococcus aureus
Haemophilus influenzaeMoraxella catarrhalisStreptococcus agalactiae
Escherichia coliProteus spp.Streptococcus pneumoniae
Klebsiella aerogenesPseudomonas aeruginosaStreptococcus pyogenes

The following atypical bacteria, viruses, and antimicrobial resistance genes are reported qualitatively:

Atypical Bacteria
Chlamydia pneumoniaeLegionella pneumophilaMycoplasma pneumoniae
Viruses
AdenovirusHuman Rhinovirus/EnterovirusParainfluenza Virus
CoronavirusInfluenza ARespiratory Syncytial Virus
Human MetapneumovirusInfluenza B
Antimicrobial Resistance Genes
CTX-MNDMmecA/C and MREJ
IMPOXA-48-like
KPCVIM

The detection and identification of specific viral and bacterial nucleic acids, as well as the estimation of relative abundance of nucleic acid from common bacterial analytes, within specimens collected from individuals exhibiting signs and/or symptoms of a respiratory infection, aids in the diagnosis of lower respiratory infection if used in conjunction with other clinical and epidemiological information. The results of this test should not be used as the sole basis for diagnosis, treatment, or other patient management decisions.

Negative results in the setting of a respiratory illness may be due to infection with pathogens that are not detected by this test, pathogens below the limit of detection, or in the case of bacterial analytes, present at levels below the lowest reported 10/4 copies/mL bin. Detection of analytes does not rule out coinfection with other organisms: the agent(s) detected by the FilmArray Pneumonia Panel may not be the definite cause of disease. Additional laboratory testing (e.g. bacterial and viral culture, immunofluorescence, and radiography) may be necessary when evaluating a patient with possible lower respiratory tract infection.

Detection of bacterial nucleic acid may be indicative of colonizing or normal respiratory flora and may not indicate the causative agent of pneumonia. Semi-quantitative Bin (copies/mL) results generated by the FilmArray Pneumonia Panel are not equivalent to CFU/mL and do not consistently correlate with the quantity of bacterial analytes compared to CFU/mL. For specimens with multiple bacteria detected, the relative abundance of nucleic acids (copies/mL) may not correlate with the relative abundance of bacteria as determined by culture (CFUmL). Clinical correlation is advised to determine significance of semi-quantitative Bin (copies/mL) for clinical management.

The antimicrobial resistance gene detected may or may not be associated with the agent(s) responsible for disease. Negative results for these antimicrobial resistance gene assays do not indicate susceptibility to corresponding classes of antimicrobials, as multiple mechanisms of antimicrobial resistance exist.

Antimicrobial resistance can occur via multiple mechanisms. A "Not Detected" result for a genetic marker of antimicrobial resistance does not indicate susceptibility to associated antimicrobial drugs or

8

drug classes. A "Detected" result for a genetic marker of antimicrobial resistance cannot be definitively linked to the microorganism(s) detected. Culture is required to obtain isolates for antimicrobial susceptibility testing, and FilmArray Pneumonia Panel results should be used in conjunction with culture results for determination of bacterial susceptibility or resistance.

Due to the genetic similarity between human rhinovirus and enterovirus, the test cannot reliably differentiate them. A positive Rhinovirus result should be followed up using an alternate method (e.g., cell culture or sequence analysis) if differentiation is required.

Culture is required to identify pathogens not detected by the FilmArray Pneumonia Panel, to further speciate analytes in genus, complex, or group results if desired, to identify bacterial pathogens present below the 10^4 copies/mL bin if desired, and for antimicrobial susceptibility testing.

Device Description:

The FilmArray Pneumonia Panel is designed to simultaneously identify 26 potential pathogens of lower respiratory tract infection (LRTI) and associated antimicrobial resistance (AMR) genes from a sputumlike (induced and expectorated sputum as well as endotracheal aspirate, ETA) or bronchoalveolar lavage (BAL)-like (BAL and mini-BAL) specimens obtained from individuals with signs and/or symptoms of lower respiratory tract infection in a time (~1 hour) that allows the test results to be used in determining appropriate patient treatment and management. FilmArray Pneumonia Panel is compatible with BioFire Diagnostics' (BioFire) PCR-based in vitro diagnostic FilmArray 2.0, and FilmArray Torch systems for infectious disease testing. A specific software module (i.e. FilmArray Pneumonia Panel pouch module) is used to perform FilmArray Pneumonia Panel testing on these systems.

Bacteria - Quantitative ResultsAntimicrobial Resistance Genes
Acinetobacter calcoaceticus-baumannii complexblaCTX-M (Extended spectrum beta-lactamase (ESBL))
Enterobacter cloacae complexblaIMP (Carbapenem resistance)
Escherichia coliblaKPC (Carbapenem resistance)
Haemophilus influenzaemecA/mecC and MREJ (Methicillin resistance)
Klebsiella aerogenesblaNDM (Carbapenem resistance)
Klebsiella oxytocablaOXA48-like (Carbapenem resistance)
Klebsiella pneumoniae groupblaVIM (Carbapenem resistance)
Moraxella catarrhalis
Proteus spp.Adenovirus
Pseudomonas aeruginosaCoronavirus
Serratia marcescensHuman Metapneumovirus
Streptococcus agalactiaeHuman Rhinovirus/Enterovirus
Streptococcus pneumoniaeInfluenza A
Streptococcus pyogenesInfluenza B
Bacteria (Atypical) - Qualitative ResultsViruses
Chlamydia pneumoniaeParainfluenza Virus
Legionella pneumophilaRespiratory Syncytial Virus
Mycoplasma pneumoniae

A test is initiated by loading Hydration Solution into one port of the FilmArray pouch and a sputum-like or BAL-like sample mixed with the provided Sample Buffer into the other port of the FilmArray Pneumonia Panel pouch and placing it in a FilmArray instrument. The pouch contains all of the reagents

9

required for specimen testing and analysis in a freeze-dried format; the addition of Hydration Solution and Sample/Buffer Mix rehydrates the reagents. After the pouch is prepared, the FilmArray Software guides the user though the steps of placing the instrument, scanning the pouch barcode, entering the sample identification, and initiating the run.

The FilmArray instrument contains a coordinated system of inflatable bladders and seal points, which act on the pouch to control the movement of liquid between the pouch blisters. When a bladder is inflated over a reagent blister, it forces liquid from the blister into connecting channels. Alternatively, when a seal is placed over a connecting channel it acts as a valve to open or close a channel. In addition, electronically-controlled pneumatic pistons are positioned over multiple plungers in order to deliver the rehydrated reagents into the blisters at the appropriate times. Two Peltier devices control heating and cooling of the pouch to drive the PCR reactions and the melt curve analysis.

Nucleic acid extraction occurs within the FilmArray pouch using mechanical and chemical lysis followed by purification using standard magnetic bead technology. After extracting and purifying nucleic acids from the unprocessed sample, the FilmArray performs a nested multiplex PCR that is executed in two stages. During the filmArray performs a single, large volume, highly multiplexed reverse transcription PCR (rt-PCR) reaction. The products from first stage PCR are then diluted and combined with a fresh, primer-free master mix and a fluorescent double stranded DNA binding dye (LC Green® Plus, BioFire Diagnostics). The solution is then distributed to each well of the array. Array wells contain sets of primers designed specifically to amplify sequences internal to the PCR products generated during the first stage PCR reaction. The 2nd stage PCR, or nested PCR, is performed in singleplex fashion in each well of the array. At the conclusion of the 2nd stage PCR, the array is interrogated by melt curve analysis for the detection of signature amplicons denoting the presence of specific targets. A digital camera placed in front of the 2nd stage PCR captures fluorescent images of the PCR reactions and software interprets the data.

The FilmArray Software automatically interprets the results of each DNA melt curve analysis and combines the data with the results of the internal pouch controls to provide a test result for each organism on the panel.

Substantial Equivalence:

The FilmArray Pneumonia Panel is substantially equivalent to the Curetis Unyvero LRT Application (DEN170047), which was recently cleared on April 03, 2018 and determined to be a Class II device under the classification product code QBH.

Table 1 compares the FilmArray Pneumonia Panel to the Curetis Unyvero LRT Application and outlines the similarities and differences between the two systems.

| Element | New Device:
FilmArray Pneumonia Panel | Predicate:
Curetis Unyvero LRT Application
(DEN170047) |
|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Specimen
Types | Sputum-like (induced or expectorated
sputum, endotracheal aspirates) and BAL-
like (BAL or mini-BAL) specimens from
individuals (of all ages) with signs and/or
symptoms of lower respiratory tract
infection | Endotracheal aspirate specimens from adult
hospitalized patients with suspected lower
respiratory tract infections |
| Element | New Device:
FilmArray Pneumonia Panel | Predicate:
Curetis Unyvero LRT Application
(DEN170047) |
| Organisms
Detected | Bacteria: Acinetobacter calcoaceticus-
baumannii complex, Enterobacter cloacae
complex, Escherichia coli , Haemophilus
influenzae, Klebsiella aerogenes, Klebsiella
oxytoca, Klebsiella pneumoniae group,
Moraxella catarrhalis, Proteus spp.,
Pseudomonas aeruginosa, Serratia
marcescens, Staphylococcus aureus,
Streptococcus agalactiae, Streptococcus
pneumoniae, Streptococcus pyogenes
Atypical Bacteria: Chlamydia pneumoniae,
Legionella pneumophila, Mycoplasma
pneumoniae
Antimicrobial Resistance Genes: CTX-M,
IMP, KPC, $mecA/C$ + MREJ, NDM,
OXA48-like, VIM | Bacteria: Acinetobacter spp., Citrobacter
freundii, Enterobacter cloacae complex,
Escherichia coli, Haemophilus influenzae,
Klebsiella oxytoca, Klebsiella pneumoniae,
Klebsiella variicola, Moraxella catarrhalis,
Morganella morganii, Proteus spp.,
Pseudomonas aeruginosa, Serratia
marcescens, Staphylococcus aureus,
Stenotrophomonas maltophilia,
Streptococcus pneumoniae
Atypical Bacteria: Chlamydia pneumoniae,
Legionella pneumophila, Mycoplasma
pneumonia,
Antimicrobial Resistance Genes: CTX-M,
KPC, mecA, NDM, OXA-23, OXA-24,
OXA-48, OXA-58, TEM, VIM |
| Viruses: Adenovirus, Coronavirus, Human
Metapneumovirus, Human
Rhinovirus/Enterovirus, Influenza A,
Influenza B, Parainfluenza virus,
Respiratory Syncytial virus | | |
| Analyte | DNA/RNA | DNA only |
| Technological
Principles | Multiplex nucleic acid | Same |
| Result types | Semi-quantitative detection with bin values
(in 1-log rounded copies/mL bins) for
Bacteria
Qualitative for Atypical Bacteria,
Antimicrobial Resistance Genes, and
Viruses | Qualitative for all analytes |
| Instrumentation | FilmArray, FilmArray 2.0, or FilmArray
Torch | Unyvero System |
| Time to result | About 1 hour | About 4-5 hours |
| Reagent
Storage | Room temperature | Unknown |
| Test
Interpretation | Automated test interpretation and report
generation. User cannot access raw data. | Same |
| Controls | Two controls are included in each reagent
pouch to control for sample processing and
both stages of PCR and melt analysis. | One control is included in each cartridge to
monitor for the presence of PCR inhibitors
and enables the system to detect any failures
in the testing process. |
| User
Complexity | Moderate/Low | Same |

Table 1. Comparison of the FilmArray Pneumonia Panel and the Curetis Unyvero LRT Application)

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11

Summary of Performance Data

Clinical Performance

The clinical performance of the FilmArray Pneumonia Panel was established during a multi-center study conducted at eight geographically distinct U.S. study sites from October 2016 to July 2017. A total of 904 residual BAL (821 BAL and 83 mini-BAL) and 925 residual sputum ( 478 sputum and 447 ETA) specimens were acquired for the prospective clinical study. FilmArray Pneumonia Panel performance in BAL and mini-BAL was similar, as was performance in sputum and ETA; therefore these sample types are not stratified further in performance tables. A total of 58 BAL and 89 sputum specimens were excluded from the final data analysis. The most common reasons for specimen exclusion for both specimen types was reference culture unable to be performed, the specimen was found to not meet the inclusion criteria after the specimen had been enrolled, or the study site was unable to complete the Case Report Form (CRF). The final data set consisted of 846 BAL and 836 sputum specimens. Table 2 and Table 3 provide a summary of demographic information for the specimens included in the prospective study.

BAL
OverallSite 1ªSite 2Site 3Site 4Site 5Site 6Site 7Site 8
SexMale480 (57%)80 (59%)7 (54%)138 (55%)21 (68%)75 (61%)82 (52%)27 (61%)50 (55%)
Female366 (43%)55 (41%)6 (46%)113 (45%)10 (32%)48 (39%)76 (48%)17 (39%)41 (45%)
Age≤ 5 years23 (3%)0 (0%)5 (38%)0 (0%)15 (48%)0 (0%)3 (2%)0 (0%)0 (0%)
6 - 17 years27 (3%)0 (0%)8 (62%)0 (0%)13 (42%)0 (0%)4 (3%)1 (2%)1 (1%)
18 - 34 years70 (8%)18 (13%)0 (0%)17 (7%)3 (10%)10 (8%)10 (6%)5 (11%)7 (8%)
35 - 65 years470 (56%)78 (58%)0 (0%)152 (61%)0 (0%)70 (57%)88 (56%)27 (61%)55 (60%)
> 65 years255 (30%)38 (28%)0 (0%)82 (33%)0 (0%)43 (35%)53 (34%)11 (25%)28 (31%)
Care SettingHospitalized666 (79%)116 (86%)12 (92%)223 (89%)9 (29%)82 (67%)118 (75%)25 (57%)81 (89%)
Outpatient159 (19%)18 (13%)0 (0%)28 (11%)22 (71%)31 (25%)39 (25%)14 (32%)7 (8%)
Emergency21 (2%)1 (1%)1 (8%)0 (0%)0 (0%)10 (8%)1 (1%)5 (11%)3 (3%)
Total84613513251311231584491

Table 2. Overall and Per Site Demographic Analysis for BAL Specimens

a Subject age could not be determined for one specimen from Site 1

Table 3. Overall and Per Site Demographic Analysis for Sputum Specimens

Sputum
OverallSite 1Site 2Site 3Site 4Site 5Site 6Site 7Site 8
SexMale481 (58%)66 (59%)54 (54%)136 (56%)97 (61%)14 (82%)31 (53%)34 (74%)49 (47%)
Female355 (42%)45 (41%)46 (46%)105 (44%)61 (39%)3 (18%)28 (47%)12 (26%)55 (53%)
Age≤ 5 years138 (17%)0 (0%)49 (49%)0 (0%)80 (51%)0 (0%)0 (0%)2 (4%)7 (7%)
6 - 17 years107 (13%)0 (0%)35 (35%)0 (0%)64 (41%)0 (0%)0 (0%)2 (4%)6 (6%)
18 - 34 years86 (10%)15 (14%)16 (16%)20 (8%)13 (8%)1 (6%)6 (10%)5 (11%)10 (10%)
35 - 65 years284 (34%)51 (46%)0 (0%)133 (55%)1 (1%)6 (35%)36 (61%)20 (43%)37 (36%)
> 65 years221 (26%)45 (41%)0 (0%)88 (37%)0 (0%)10 (59%)17 (29%)17 (37%)44 (42%)
Care SettingHospitalized682 (82%)106 (95%)64 (64%)219 (91%)105 (66%)12 (71%)52 (88%)23 (50%)101 (97%)
Outpatient73 (9%)2 (2%)14 (14%)18 (7%)24 (15%)2 (12%)5 (8%)7 (15%)1 (1%)
Emergency81 (10%)3 (3%)22 (22%)4 (2%)29 (18%)3 (18%)2 (3%)16 (35%)2 (2%)

12

Sputum
OverallSite 1Site 2Site 3Site 4Site 5Site 6Site 7Site 8
Total836111100241158175946104

All specimens were evaluated with the FilmArray Pneumonia Panel at clinical study sites. Refrigerated specimen aliquots were sent to a central reference laboratory for quantitative reference culture (qRefCx) and frozen specimen aliquots were also sent to BioFire for evaluation by polymerase chain reaction (PCR)/sequencing-based comparator methods.

The reference methods used in this study were as follows:

Bacterial analytes were compared to qRefCx to evaluate sensitivity and the method was considered positive for the presence of the organism of interest if it was recovered in culture and enumerated at a level of 3162 (10^3.5) CFU/mL or greater.

Bacterial analytes were also evaluated by comparison to a single PCR assay for the organism of interest followed by a quantitative molecular assay that included sequencing (qMol) to assess FilmArray bin reporting performance. Atypical bacteria and viruses were compared to two conventional PCR assays followed by bidirectional sequencing. For specimens with an applicable bacteria detected by FilmArray, AMR genes were compared to a single PCR assay (from the specimen) followed by sequencing. A specimen was considered to be positive for an analyte if bi-directional sequencing data meeting predefined quality acceptance criteria matched organism-specific sequences deposited in the NCBI GenBank database (www.ncbi.nlm.nih.gov) with acceptable E-values. When two PCR comparator assays were used, any specimen that tested negative by both of the comparator assays was considered Negative.

Positive Percent Agreement (PPA) or Sensitivity for each analyte was calculated as 100% x (TP + FN)). True positive (TP) indicates that both the FilmArray Pneumonia Panel and the comparator method had a positive result for this specific analyte, and false negative (FN) indicates that the FilmArray Pneumonia Panel result was negative while the comparator result was positive. Negative Percent Agreement (NPA) or Specificity was calculated as 100% x (TN + FP)). True negative (TN) indicates that both the FilmArray Pneumonia Panel and the comparator method had negative results, and a false positive (FP) indicates that the FilmArray Pneumonia Panel result was positive but the comparator result was negative. The exact binomial two-sided 95% confidence interval was calculated. Samples for which false positive and/or false negative results (i.e., discrepant results) were obtained when comparing the FilmArray Pneumonia Panel results to the comparator method results were further investigated. The discrepancy investigations were primarily performed as follows: for discrepancies between the Pneumonia Panel and reference culture for bacterial analytes, discrepancies were first examined to see if qRefCx or FilmArray had observed the analyte but reported it as "negative" or "Not Detected" because it was below the detection threshold. If this did not resolve the discrepancy, the results of qMol testing were considered. And if these methods still did not resolve the discrepancy, it was then investigated in the same manner as other analytes that used molecular comparator (i.e. using multiple additional molecular assays followed by sequence analysis). The results of SOC testing were also considered. The prospective clinical study results are summarized in Table 5 for BAL and sputum specimens, respectively.

BAL
AnalyteReferenceSensitivity/PPASpecificity/NPA

13

MethodTP/(TP + FN)%95%CITN/(TN + FP)%95%CI
Bacteria
Acinetobacter calcoaceticus-baumannii complexbqRefCx0/0-839/84699.298.3-99.6%
Klebsiella aerogenescqRefCx6/785.748.7-97.4%832/83999.298.3-99.6%
Enterobacter cloacae complexdqRefCx11/1291.764.6-98.5%822/83498.697.5-99.2%
Escherichia colieqRefCx12/1210075.8-100%826/83499.098.1-99.5%
Haemophilus influenzaefqRefCx10/1010072.2-100%764/83691.489.3-93.1%
Klebsiella oxytocagqRefCx2/210034.2-100%835/84498.998.0-99.4%
Klebsiella pneumoniae grouphqRefCx15/1510079.6-100%819/83198.697.5-99.2%
Moraxella catarrhalisiqRefCx0/0-817/84696.695.1-97.6%
Proteus spp.jqRefCx5/510056.6-100%837/84199.598.8-99.8%
Pseudomonas aeruginosakqRefCx36/3610090.4-100%772/81095.393.6-96.6%
Serratia marcescenslqRefCx6/610061.0-100%834/84099.398.5-99.7%
Staphylococcus aureusmqRefCx46/4797.988.9-99.6%729/79991.289.1-93.0%
Streptococcus agalactiaenqRefCx1/1100-821/84597.295.8-98.1%
Streptococcus pneumoniaeoqRefCx5/510056.6-100%817/84197.195.8-98.1%
Streptococcus pyogenespqRefCx2/210034.2-100%838/84499.398.5-99.7%
Atypical Bacteria
Chlamydia pneumoniaeqPCR/Seq0/0--844/84599.999.3-100%
Legionella pneumophilaPCR/Seq2/210034.2-100%833/83310099.5-100%
Mycoplasma pneumoniaerPCR/Seq3/310043.9-100%841/84299.999.3-100%
Viruses
AdenovirusPCR/Seq8/810067.6-100%837/83710099.5-100%
CoronavirussPCR/Seq18/2185.765.4-95.0%810/82398.497.3-99.1%
Human MetapneumovirustPCR/Seq8/810067.6-100%836/83799.999.3-100%
Human Rhinovirus/EnterovirusuPCR/Seq52/5496.387.5-99.0%771/78298.697.5-99.2%
Influenza AvPCR/Seq10/1010072.2-100%830/83399.698.9-99.9%
Influenza BwPCR/Seq5/683.343.6-97.0%837/83899.999.3-100%
Parainfluenza VirusxPCR/Seq16/1888.967.2-96.9%824/82699.899.1-99.9%
Respiratory Syncytial VirusPCR/Seq3/310043.9-100%841/84110099.5-100%

" The performance measures of sensitivity and specificity only refer to the bacterial analytes for which the rold-standard of qRefCx was used as the reference method. Performance measures of PPA and NPA refer to all other analytes, for which PCR sequencing assays were used as comparator methods.

b Evidence of ACB complex was found in all seven FP speciment below 10°3.5 CFUmL by gRefCx and six were detected by oMol.

f K. aerogenes was identified in the single FN specimen in SOC culture. Evidence of K. aevogeners four were enumerated below 10^3.5 CFU/mL by qRefCx and three were detected by qMol.

4 E. cloacae complex was observed in the single FN specimen below the FilmArray Pheumonia Panel. Evidence of E. cloacae complex was found in all 12 FP speciment six were enumerated below 10°3.5 CFUmL by qRefCx, five were detected using an additional molecular method.

" Evidence of £. coli was found in all eight FP speciments below 10°3.5 CFUmL by qRefCx and two were detected by qMol.

'Evidence of H. influenzae was found in all 72 FP specimented below 10'3.5 CFU/mL by qRefCx, 56 were detected by aMol, eight were detected using an additional molecular method, and one was identified in SOC culture.

6 Evidence of K. oxyloca was found in all nine FP specimented below 10°3.5 CFUmL by qReCx, five were detected by qMol, and one was detected using an additional molecular method.

4 Evidence of K. pneumoniae group was found in all 12 FP specimented below 10°3.5 CFUmL by qRefCx, four were detected by qMol, and one was detected using an additional molecular method.

' Evidence of M. catarrhalis was found in all 29 FP speciments two were enumerated below 10'3.5 CFUmL by qRefCx, 25 were detected by qMol, and two were detected using an additional molecular method.

I Evidence of Proteus spp. was found in all four FP specimented below 10°3.5 CFUmL by qRefCx and one was detected by aMol.

  • Evidence of P. aeruginosa was found in all 38 FP speciments below 10°3.5 CFU/mL by qRefCx, 16 were detected by qMol, and three were detected using an additional molecular method.

' Evidence of S. marcescens was found in all six FP specimented below 10'3.5 CFU/mL by qRefCx and two were detected by gMol.

14

" S. aureus was detected in the single FN specimen using an motecular method. Evidence of S. aureus was found in 6970 FP specimens; 29 were enumerated below 10'3.5 CFUmL by qRefCx, 30 were detected using an additional molecular method, and two were identified in SOC culture.

" Evidence of S. agalactiae was found in all 24 FP speciments below 10'3.5 CFUmL by qRefCx, 13 were detected by aNol, and four were detected using an additional molecular method.

° Evidence of S. pneumoniae was found in all 24 FP specimented below 10°3.5 CFUmL by qRefCx, 18 were detected by qMol, and one was detected using an additional molecular method.

P Evidence of S. pyogenes was found in all six FP specimented below 10'3.5 CFUmL by qRefCx, three were detected by aNol, and one was detected using an additional molecular method.

9 The single FP specimen was negative for C. pneumoniae when tested with additional mothods during discrepancy investigation.

" The single FP specimen was negative for M. pneumoniae when methods during discrepancy investigation.

  • CoV was detected in 2/3 FN and 8/13 FP specimens using an additional molecular method.

' The single FP specimen was negative for hMPV when tested with additional methods during discrepancy investigation.

" HRVEV was detected in both FN speciment molecular method. HRV/EV was detected in 8/11 FP specimens during discepancy investigation; seven were detected using an additional mothod and one was detected upon FilmArray Pneumonia Panel retest.

Y FluA was detected in 2/3 FP specimens using an additional molecular method.

" FuB was detected in the single FN specimen upon FilmArray Pheumonia Panel retest. FluB was detected in the an addrional molecular method.

  • PIV was detected in both FN and both FP specimens using an additional molecular method.

Table 5. FilmArray Pneumonia Panel Clinical Performance Summary for Sputum Specimens®

Sputum
AnalyteReference
MethodTP/(TP +
FN)%95%CITN/(TN +
FP)%95%CI
Bacteria
Acinetobacter calcoaceticus-baumannii complexbqRefCx10/1190.962.3-98.4%807/82597.896.6-98.6%
Klebsiella aerogenescqRefCx3/475.030.1-95.4%823/83298.998.0-99.4%
Enterobacter cloacae complexdqRefCx11/1291.764.6-98.5%803/82497.596.1-98.3%
Escherichia colieqRefCx23/2495.879.8-99.3%787/81296.995.5-97.9%
Haemophilus influenzaefqRefCx16/1888.967.2-96.9%727/81888.986.5-90.9%
Klebsiella oxytocagqRefCx9/910070.1-100%817/82798.897.8-99.3%
Klebsiella pneumoniae grouphqRefCx21/2391.373.2-97.6%769/81394.692.8-95.9%
Moraxella catarrhalisiqRefCx5/510056.6-100%761/83191.689.5-93.3%
Proteus spp.jqRefCx15/1510079.6-100%813/82199.098.1-99.5%
Pseudomonas aeruginosakqRefCx103/10697.292.0-99.0%673/73092.290.0-93.9%
Serratia marcescenslqRefCx26/2796.381.7-99.3%782/80996.795.2-97.7%
Staphylococcus aureusmqRefCx111/11299.195.1-99.8%631/72487.284.5-89.4%
Streptococcus agalactiaenqRefCx9/910070.1-100%793/82795.994.3-97.0%
Streptococcus pneumoniaeoqRefCx16/1610080.6-100%785/82095.794.1-96.9%
Streptococcus pyogenespqRefCx6/610061.0-100%825/83099.498.6-99.7%
Atypical Bacteria
Chlamydia pneumoniaePCR/Seq0/0--835/83510099.5-100%
Legionella pneumophilaqPCR/Seq0/10-826/82610099.5-100%
Mycoplasma pneumoniaerPCR/Seq7/887.552.9-97.8%827/82710099.5-100%
Viruses
AdenovirussPCR/Seq13/1776.552.7-90.4%815/81799.899.1-99.9%
CoronavirustPCR/Seq28/3287.571.9-95.0%796/80299.398.4-99.7%
Human MetapneumovirusuPCR/Seq20/2195.277.3-99.2%812/81399.999.3-100%
Human Rhinovirus/EnterovirusvPCR/Seq96/9610096.2-100%717/73098.297.0-99.0%
Influenza AwPCR/Seq13/1310077.2-100%819/82299.698.9-99.9%
Influenza BxPCR/Seq12/1210075.8-100%821/82399.899.1-99.9%

15

Sputum
AnalyteReference
MethodSensitivity/PPA
TP/(TP + FN)%95%CISpecificity/NPA
TN/(TN + FP)%95%CI
Parainfluenza VirusyPCR/Seq28/2996.682.8-99.4%804/80699.899.1-99.9%
Respiratory Syncytial ViruszPCR/Seq43/4310091.8-100%787/79199.598.7-99.8%

" The performance measures of sensitivity and specificity only refer to the bacterial analytes for which the gold-standard of qRefCx was used as the reference method. Performance measures of PPA and NPA refer to all other analytes, for which PCR/sequencing assays were used as comparator methods.

o The isolate recovered from the single FN specific by qReCx; molecular testing of the isolate identified it as Pseudononas fluvescens during discrepancy investigation. Evidence of ACB complex was found in all 18 FP specimens; 15 were detected using an additional molecular method, and one was identified in SOC culture.

6 The isolate recovered from the single FN specifics by qRefCx; molecular testing of the isolate identified it as Hafnia paralvei during discrepancy investigation. Evidence of K. aerogenes was found in all nine FP specimens; three were enumerated below 10°3,5 CFU/mL by qRefCx, five were detected by qMol, and one was detected using an additional molecular method.

4 E. cloace complex was detected in the single FN specifical molecular method. Evidence of E. cloace complex was found in all 21 FP specimens; four were enumerated below 103.5 CFUlmL by qRefCx, 16 were was detected using an additional molecular method.

s E. coli was observed in the single FN speciment of 4 bin by the FilmArray Preumonia Panel. Evidence of E. coli was found in all 25 FP specimens; six were enumerated below 10°3.5 CFUmL by qRefCx, 14 were detected using an additional molecular method.

f H. influenzae was detected in 1/2 FN specimens by gMol. The isolate recovered from the other FN specimen was misidentified by qRefCx; molecular testing of the isolate identified it as Heenophicus during discrepancy investigation. Evidence of H. influenzae was found in all 91 PP specimens; four were enumerated below 10°3.5 CFUmL by qRefCx, 78 were detected using an additional molecular method, and two were identified in SOC culture.

4 Evidence of K. oxy/oca was found in all 10 FP speciments three were enumerated below 10°3.5 CFU/mL by gRefCx, five were detected by qMol, and two were detected using an additional molecular method.

4 K. pneumoniae group was detected in 1/2 FN speared to be a result of a specimen swap at the central reference laboratory. Evidence of K. pneumoniae group was found in 4344 FP speciments 15 were enumerated below 103.5 CFUmL by gRefCx, 21 were detected by qMol, and seven were detected using an additional molecular method.

' Evidence of M. catarrhalis was found in all 70 FP speciments one was enumerated below 10°3.5 CFU/mL by qRefCx, 63 were detected by aMol, five were detected using an additional molecular method, and one was identified in SOC culture.

I Evidence of Protens spp. was found in all eight FP specimented below 10°3.5 CFUmL by qReCx, four were detected by aMol, and two were detected using an additional molecular method.

k P. aeruginosa was observed in 1/3 FN speciment be 10% bin by the FilmArray Pheumonia Panel. The isolates recovered from the other two FN specimens were misidentified by gRefCx; molecular testing of the isolates identificans and the other as Pseudononas fluorescens during discrepancy investigation of P. aeruginosa was found in all 57 FP speciments 21 were enumerated below 10°3.5 CFUmL by qRefCx, 33 were detected by qMol, two were detected using an additional molecular method, and one was identified in SOC culture.

1 S. marcescens was observed in the single the 10 4 bin by the FilmArray Preumonia Panel. Evidence of S. marcescens was found in 26/27 FP specimens; seven were enumerated below 10°3.5 CFUmL by qRefCx, 16 were detected using an additional molecular method.

™ S. aureus was observed in the single FN speciment below the FilmArray Preumonia Panel. Evidence of S. aureas was found in all 93 PP specimens; 43 were enumerated below 10°3.5 CFUmL by qReCx, 43 were detected using an additional molecular nethod, and four were identified in SOC culture.

" Evidence of S. gealactive was found in all 34 FP specimented below 10°3.5 CFUmL by qRefCx, 24 were detected by qMol, and five were detected using an additional molecular method

° Evidence of S, pneumoniae was found in all 35 FP specimented below 10°3.5 CFUmL by gRefCx and 34 were detected by gMol.

P Evidence of S. pyogenes was found in all five FP speciments four were detected using an additional molecular method.

9 L. pneumophila was detected in the single FN specimen using an additional molecular method.

T The single FN specimen was negative for M. pneumoniae when tested during discrepancy investigation.

5 AdV was detected in all four FN and 1/2 FP specimens using an additional molecular method.

1 CoV was detected in all four FN and 3/6 FP specimens using an additional molecular method.

" hMPV was detected in the single FN specimen using an method. The single FP specimen was negative for MPV when tested during discrepancy investigation.

  • HRVEV was detected in 12/13 FP specimens during discrepancy investigation; 11 were detected using an additional method and one was detected upon FilmArray Pneumonia Panel retest.

  • FluA was detected in all three FP specimens using an additional molecular method.

  • Both FP specimens were negative for FluB when tested with additional mothods during discrepancy investigation.

y PIV was detected in the single FN and 1/2 FP specimens using an additional molecular method.

16

2 RSV was detected in all four FP specimens using an additional molecular method.

A total of 156 BAL specimens and 295 sputum specimens received a FilmArray Pneumonia Panel Detected result for at least one applicable gram-negative bacterium on the panel and reported results for CTX-M, IMP, KPC, NDM, OXA-48-like, and VIM. A total of 116 BAL specimens and 204 sputum specimens received a FilmArray Pneumonia Panel Staphylococcus aureus Detected result and reported results for mecA/C and MREJ. Performance of the Pneumonia Panel AMR gene assays was calculated by comparing results of qMol direct from these specimens and is shown in Table 6 (five BAL and four sputum specimens were excluded from qMol analysis due to invalid comparator results).

BALSputum
PPANPAPPANPA
AnalyteTP/
(TP +
FN)%95%CITN/
(TN +
FP)%95%CITP/
(TP +
FN)%95%CITN/
(TN +
FP)%95%CI
CTX-
Mb6/785.748.7-
97.4%144/14410097.4-
100%8/1080.049.0-
94.3%280/28199.698.0-
99.9%
IMP0/0--151/15110097.5-
100%0/0--291/29110098.7-
100%
KPCc2/210034.2-
100%148/14999.396.3-
99.9%7/710064.6-
100%284/28410098.7-
100%
mecA/C
and
MREJd40/4588.976.5-
95.2%64/7091.482.5-
96.0%94/9895.990.0-
98.4%91/10487.579.8-
92.5%
NDMe0/10-149/15099.396.3-
99.9%0/0--291/29110098.7-
100%
OXA-
48-
like0/0--151/15110097.5-
100%0/0--291/29110098.7-
100%
VIMf0/0--151/15110097.5-
100%1/1100-289/29099.798.1-
99.9%

Table 6. FilmArray Pneumonia Panel Clinical Performance Summarator method: qMol direct from snecimen)ª

3 Performance in this summary table is calculated when any applicable organism is detected in the sample.

· CTX-M was detected in the single FN BAL and 1/2 FN sputum specifical molecular method. The single FP sputum specimen was negative for CTX-M when tested during discrepancy investigation. None of the applicable isolates identified by the FilmArray or qRefCx from these specimens had evidence of ESBL activity or CTX-M presence.

" KPC was detected in the single FP BAL specimen using an additional mothod; the isolate recovered from this speciment (A baumanni) exhibited carbapenem resistance but did not carry KPC.

4 Eividence of nec.4C and or SCCnec casette elements was found in all five FN BAL and all four FN sputum specimens by an additional molecular method: three of these also had a MRSA isolate recovered via qRefCx or SOC culture. Evidence of mec.//C and/or SCC.nec casette elements was found in 5/6 FP BAL and all 13 FP sputum speciments nisolate recovered via qRefCx or SOC culture, and nine additional specimens had evidence of mecA/C and/or SCCmec cassette genetic elements by an additional molecular method.

  • NDM was detected in the single FN BAL specimen using an method: P. aeruginosa was recovered from the specimen and was resistant to carbapens but carred only KPC. The single FP BAL specimen was negative for NDM when tested with additional molecular methods during discepancy investigation.

f The single FP sputum specimen was negative for VIM when tested with additional methods during discrepancy investigation.

qRefCx isolated one or more applicable gram-negative bacteria from 127 of the 156 BAL specimens and 230 of the 295 sputum specimens that received a FilmArray Pneumonia Panel Detected result for an applicable gram-negative bacterium. Additionally, qRefCx isolated S. aureus from 75 of 116 BAL specimens and 154 of 204 sputum specimens that received a FilmArray Pneumonia Panel Staphylococcus aureus Detected result. The method used to assess correlation of the CTX-M, IMP, KPC, NDM, OXA-48-like, and VIM results (Table 7 and Table 8) or the mecA/C and MREJ results (Error! Reference source not found. and Error! Reference source not found.) reported in the specimen by the FilmArray Pneumonia Panel to identification of the cultured isolates from

17

that particular specimen was one conventional PCR assay followed by bidirectional sequencing, performed directly on the isolate.

18

BAL
Applicable Bacteria ResultNCTX-MIMPKPCNDMVIMOverall
(any resistance gene)
(FilmArray)PPANPAPPANPAPPANPAPPANPAPPANPAPPANPA
Overall
(any applicable bacteria
Detected)ª1274/4
(100%)121/123
(98.4%)0/0
(-)127/127
(100%)1/1
(100%)124/126
(98.4%)0/0
(-)127/127
(100%)0/0
(-)127/127
(100%)5/5
(100%)
56.6-100%118/122
(96.7%)
91.9-98.7%
Acinetobacter calcoaceticus-
baumannii complex0------------
Enterobacter cloacae complex90/0
(-)9/9
(100%)0/0
(-)9/9
(100%)0/0
(-)9/9
(100%)0/0
(-)9/9
(100%)0/0
(-)9/9
(100%)0/0
(-)9/9
(100%)
Escherichia coli124/4
(100%)8/8
(100%)0/0
(-)12/12
(100%)0/0
(-)12/12
(100%)0/0
(-)12/12
(100%)0/0
(-)12/12
(100%)4/4
(100%)8/8
(100%)
Klebsiella aerogenes70/0
(-)7/7
(100%)0/0
(-)7/7
(100%)0/0
(-)7/7
(100%)0/0
(-)7/7
(100%)0/0
(-)7/7
(100%)0/0
(-)7/7
(100%)
Klebsiella oxytoca20/0
(-)2/2
(100%)0/0
(-)2/2
(100%)0/0
(-)2/2
(100%)0/0
(-)2/2
(100%)0/0
(-)2/2
(100%)0/0
(-)2/2
(100%)
Klebsiella pneumoniae group140/0
(-)14/14
(100%)0/0
(-)14/14
(100%)0/0
(-)14/14
(100%)0/0
(-)14/14
(100%)0/0
(-)14/14
(100%)0/0
(-)14/14
(100%)
Proteus spp.60/0
(-)6/6
(100%)0/0
(-)6/6
(100%)0/0
(-)6/6
(100%)0/0
(-)6/6
(100%)0/0
(-)6/6
(100%)0/0
(-)6/6
(100%)
Pseudomonas aeruginosa430/0
(-)42/43
(97.7%)0/0
(-)43/43
(100%)0/0
(-)43/43
(100%)0/0
(-)43/43
(100%)0/0
(-)43/43
(100%)0/0
(-)42/43
(97.7%)
Serratia marcescens60/0
(-)6/6
(100%)0/0
(-)6/6
(100%)0/0
(-)6/6
(100%)0/0
(-)6/6
(100%)0/0
(-)6/6
(100%)0/0
(-)6/6
(100%)
Polymicrobial specimens280/0
(-)27/28ᵇ
(96.4%)0/0
(-)28/28
(100%)1/1ᶜ
(100%)25/27ᵈ
(92.6%)0/0
(-)28/28
(100%)0/0
(-)28/28
(100%)1/1
(100%)24/27
(88.9%)

Table 7. CTX-M, IMP, KPC, NDM, and VIM Performance Table (PCR/seq on cultured isolate(s) from BAL specimens)

s An additional nine specimes had no applicable beceria is that one or more applicable bated by ReCla, no resisted in the culture(s) by PCR/seq from these specimens

  • One specimen E. cloace complex and K. preumonize group detected by qRefC: CTX-M was not identified in this isolate by PCV-seg)

· E. cloacae complex and P. aeruginosa detected by FilmArray and isolated by qRefCx (KPC identified from the E. cloacae isolate by PCR/seq)

^ One specimen A. calcused annamic envol dected by Film Aray (A. calcucelous-baumanni complex isolated by qReC.; KC was not deathed in this istage by PCR sept your contined one specimen Proteus spp. and P. aeruginosa detected by FilmArray (P. aeruginosa isolated by qRefCx; KPC was not identified in this isolated by PCR/seq)

Table 8. CTX-M, IMP, KPC, NDM, and VIM Performance Table (PCR/seq on cultured isolate(s) from sputum specimens)

Sputum
Applicable Bacteria ResultNCTX-MIMPКРСNDMVIMOverali
(any resistance gene)
PPANPAPPANPAPPANPAPPANPAPPANPAPPANPA
Overall
(any applicable bacteria
Detected)ª2303/4
(75.0%221/226
(97.8%)1/1
(100%)229/229
(100%)રાજ
(83.3%223/224
(99.6%)0/0
(-)230/230
(100%)1/1
(100%229/229
(100%)9/11 P
(81.8%)
52.3-94.9%214/219
(97.7%)
194.8-99.0%
Acinetobacter calcoaceticus-
baumannii complexરે દ0/0
(-)ર્સ્ટ
(100%)0/0
(-)ર્ટ રિ
(100%)0/0
(-)ર્રાર
(100%)0/0
(-)ર્ટાર
(100%)0/0
(-)ર્ રાર
(100%)0/0
(-)ર્ રાજ
(100%)
Enterobacter cloacae complex7d0/0
(-)7/7
(100%)0/0
(-)7/7
(100%)0/0
(-)7/7
(100%)0/0
(-)7/7
(100%)0/0
(-)7/7
(100%)0/0
(-)7/7
(100%)
Escherichia coli101/1
(100%)9/9
(100%)0/0
(-)10/10
(100%)0/0
(-)10/10
(100%)0/0
(-)10/10
(100%)0/0
(-)10/10
(100%)1/1
(100%)9/9
(100%)
Klebsiella aerogenes0/0
(-)3/3
(100%)0/0
(-)3/3
(100%)0/0
(-)3/3
(100%)0/0
(-)3/3
(100%)0/0
(-)3/3
(100%)0/0
(-)3/3
(100%)
Klebsiella oxytocaব0/0
(-)4/4
(100%)0/0
(-)4/4
(100%)0/0
(-)4/4
(100%)0/0
(-)4/4
(100%)0/0
(-)4/4
(100%)0/0
(-)4/4
(100%)

19

Sputum
Applicable Bacteria ResultNCTX-MIMPKPCNDMVIMOverall
(any resistance gene)
PPANPAPPANPAPPANPAPPANPAPPANPAPPANPA
Klebsiella pneumoniae group211/1
(100%)20/20
(100%)0/0
(-)21/21
(100%)1/1
(100%)20/20
(100%)0/0
(-)21/21
(100%)0/0
(-)21/21
(100%)2/2
(100%)19/19
(100%)
Proteus spp.60/0
(-)6/6
(100%)0/0
(-)6/6
(100%)0/0
(-)6/6
(100%)0/0
(-)6/6
(100%)0/0
(-)6/6
(100%)0/0
(-)6/6
(100%)
Pseudomonas aeruginosa680/1
(0%)65/67
(97.0%)0/0
(-)68/68
(100%)0/1
(0%)67/67
(100%)0/0
(-)68/68
(100%)0/0
(-)68/68
(100%)0/2
(0%)64/66
(97.0%)
Serratia marcescens14c0/0
(-)14/14
(100%)1/1
(100%)13/13
(100%)0/0
(-)14/14
(100%)0/0
(-)14/14
(100%)0/0
(-)14/14
(100%)1/1
(100%)13/13
(100%)
Polymicrobial specimens921/1f
(100%)88/91g
(96.7%)0/0
(-)92/92
(100%)4/4h
(100%)87/88
(98.9%)0/0
(-)92/92
(100%)1/1i
(100%)91/91
(100%)5/5
(100%)84/87
(96.6%)

" An additional 19 specimens had no applicable bateria december out had one or more applicable baceria issubted by PCR/sen in one speciment (E. col isolated by qRefCx), but no other resistance markers were identified in the cultured isolate(s) by PCR/seq from these specimens

b One specimen had presence of dual AMR genes (KPC and VIM)

& An A. calcoaceticus-baumannii isolate was not recovered by qRefCx for one specimen

d An E. cloacae isolate was not recovered by qRefCx for one specimen

& A S. marcescens isolate was not recovered by qRefCx for one specimen

1 E. coli and P. aeruginosa detected by FilmArray and isolated by qRefCx (CTX-M identified in the E. coli isolate by PCR/seq)

€ One specimen 4. calcoacionanii compex, Yovear spp, and P. aenginna detected by FilmArray K. memonine group and P. cerginoa isolated by clearly of C. (CTA-M va no icentified in either of these is the specifier. E. coli, K. premoniae group, and P. aernejmon issulted by Red.x. (CTX-M van it itentified in ether of these is and by PCR sec); one pociment Protection by Film-tray (P . arruginosa isolated by qReal in this isolated by PCRC .;

  • One specimen E. coli and K. premonite group control of ich KPC identified in the K. premonice issuate by PCRscaliner P. arrescencededed by Film Arrey and issiated in the S. narcesers isolate by P.Oser), ore speciment i concertions in concerners . K. cergences . K. prevense costed by Finherey (4 calcaceticas-bamania, and P. ceregional solated by ReC.x. KP Cidentified in the K. yeemmine isolate by PROS. (10 P. excentral A. calcoaceticas-bannanii compex. preumoniae group, Poteus spp. and P. aerginosa detected by Him-M. permonice and P. aernemoniae isolated by PCRSee)

'One specimen K. pneunoniae group and P. aeruginosa isolated by qRefCx; KPC was not identified in this isolate by PCRseq)

I One speinen A. calcoachines of comprise goup, and P. aenginoa detested by Elmhery (. calconcericas-bannami), K. memoriae, and P. enemoniae, and P. enemoniae, and P. enement qRefCx; VIM identified in the P. aeruginosa isolate by PCR/seq)

qRefCx isolated one or more applicable gram-negative bacteria from 79 of the 94 BAL specimens that received a FilmArray Preumonia Panel for an applicable gram-negative bacterium for OXA-48-1ike. The method used to assess correlation of the OXA-48-1ke results (Table 10) reported in the specimen by the FilmArray Preumonia Panel to identification of the gene in the cultured isolates from that specimen was one conventional PCR assay followed by bidirectional sequencing, performed directly on the isolate.

BAL
Applicable Bacteria ResultPositive Percent AgreementNegative Percent Agreement
TP/(TP + FN)%95%CITN/(TN + FP)%95%CI
Overall
(any applicable bacteria Detected)0/0--79/7910095.4-100%
Enterobacter cloacae complex0/0--10/1010072.2-100%
Escherichia coli0/0--13/1310077.2-100%

Table 9. OXA-48-like Performance Table (PCR/seg on cultured isolate(s) from BAL specimens)

20

BAL
Positive Percent AgreementNegative Percent Agreement
Applicable Bacteria ResultTP/(TP + FN)%95%CITN/(TN + FP)%95%CI
Klebsiella aerogenes0/0--7/710064.6-100%
Klebsiella oxytoca0/0--3/310043.9-100%
Klebsiella pneumoniae group0/0--15/1510079.6-100%
Proteus spp.0/0--6/610061.0-100%
Serratia marcescens0/0--8/810067.6-100%
Polymicrobial specimens0/0--17/1710081.6-100%

Table 10. OXA-48-like Performance Table (PCR/seq on cultured isolate(s) from sputum specimens)

Sputum
Applicable Bacteria ResultPositive Percent AgreementNegative Percent Agreement
TP/(TP + FN)%95%CITN/(TN + FP)%95%CI
Overall
(any applicable bacteria Detected)0/0--131/13110097.2-100%
Enterobacter cloacae complex0/0--9/910070.1-100%
Escherichia coli0/0--17/1710081.6-100%
Klebsiella aerogenes0/0--4/410051.0-100%
Klebsiella oxytoca0/0--5/510056.6-100%
Klebsiella pneumoniae group0/0--25/2510086.7-100%
Proteus spp.0/0--9/910070.1-100%
Serratia marcescens0/0--25/2510086.7-100%
Polymicrobial specimens0/0--37/3710090.6-100%

21

qRefCx isolated S. avreus from 75 of 116 BAL specimens that received a FilmArray Pneumonia Pagel Staphylococus aureas Detected result. The method used to assess correlation of the mec4/C and MREJ results (1) reported in the specimen by the FilmArray Pneumonia Panel to identification of the gene in the cultured isolates from that particular specimen was one conventional PCR assay followed by bidirectional sequencing, performed directly on the isolate.

BAL
S. aureus
mecA/C and MREJqRefCx: S. aureus
PCR/seq: mecA/C
Org+ / Res+Org+ / Res-Org -Total
FilmArray
ResultOrg+ / Res+1922546
Org+ / Res-1244570
Org -01729730
Total2027799846
PerformanceAgreement%95%CI
Org+ / Res+19/2095.0%76.4-99.1%
Org+ / Res-24/2788.9%71.9-96.1%
Org -729/79991.2%89.1-93.0%
InterpretationPPANPAPrevalence
MRSA19/20
(95.0%)799/826
(96.7%)46/846
(5.4%)
MSSA24/27
(88.9%)773/819
(94.4%)70/846
(8.3%)
S. aureus46/47
(97.9%)729/799
(91.2%)116/846
(13.7%)

from BAL specimens)

Table 11 mec4 C and MREJ 3:3 Performance Table (dres) Table 12 mec4 C and MREJ 3:3 Performance Table (gRefor & PCRseq on cultured isolate(s) from sputum specimens)

Sputum
S. aureus
mecA/C and MREJqRefCx: S. aureus
PCR/seq: mecA/C
FilmArray ResultOrg+ / Res+Org+ / Res-Org -Total
FilmArray
ResultOrg+/Res+58445107
Org+/ Res-0494897
Org -01631632
Total5854724836
PerformanceAgreement%95%CI
Org+/Res+58/58100%93.8-100%
Org+/ Res-49/5490.7%80.1-96.0%
Org -631/72487.2%84.5-89.4%
InterpretationPPANPAPrevalence
MRSA58/58
(100%)729/778
(93.7%)107/836
(12.8%)
MSSA49/54
(90.7%)734/782
(93.9%)97/836
(11.6%)
S. aureus111/112
(99.1%)631/724
(87.2%)204/836
(24.4%)

FilmArray Pneumonia Panel CTX-M reporting was also compared to the standed spectrum (B-actamase (ESBL) activity testing methods performed in conjunction with qRefCx. Standard phenotypic ESBL activity was only reported for E. coli and Klebsiella spp. by the central reference laboratory.

BioFire Diagnostics 510(k) FilmArray Pneumonia Panel

22

Of the 156 BAL specimens that received a FilmAray Panel Detected result for at least one applicable gram-negative bacterium on the panel, 53 specimens received a Detected result for E. coli, K. pneumoniae, qRefCx isslated E. coli, K. axytoca, and or K. pneumoniae from 43 of these specimens of the 295 sputum specimens that received a FilmArray Pheumonia Panel for at least one applicable gram-negative bacterium on the panel, 114 speciment received a Detected result for E. coli, K. meumoniae; qRefCx isolated E. coli, K. or:toca, and/or K. pneumoniae from 71 of these specimens. The correlation between of CTX-M in a particular specimen as compared to the phenotypic AST results of isolates recovered from the stratified by each applicable associated organism in Table 13 and Table 14Error! Reference source not found..

BAL
Positive Percent AgreementNegative Percent Agreement
Applicable Bacteria ResultTP/(TP + FN)%95%CITN/(TN + FP)%95%CI
Overall
(any applicable bacteria Detected)4/580.037.6-96.4%38/3810090.8-100%
Escherichia coli4/410051.0-100%11/1110074.1-100%
Klebsiella oxytoca0/0--5/510056.6-100%
Klebsiella pneumoniae group0/10-17/1710081.6-100%
Polymicrobial specimens0/0--5/510056.6-100%

Table 13. CTX-M Performance Table (comparison to phenotypic AST methods for BAL specimens)

Table 14. CTX-M Performance Table (comparison to phenotypic AST methods for sputum specimens)

Sputum
Positive Percent AgreementNegative Percent Agreement
Applicable Bacteria ResultTP/(TP + FN)%95%CITN/(TN + FP)%95%CI
Overall
(any applicable bacteria Detected)4/757.125.1-84.2%63/6498.491.7-99.7%
Escherichia coli2/366.720.8-93.9%17/1710081.6-100%
Klebsiella oxytoca0/0--9/910070.1-100%
Klebsiella pneumoniae group1/250.0-26/2796.381.7-99.3%
Polymicrobial specimens1/2a50.0-11/1110074.1-100%

One specimen E. col and K. axyloca detect by ReC. (ESBL activity identified in the E. coli isolate by gRefC. AST); one speciment E. coli and K. memonize group detected by FilmArray (E. coli isolated by qRefCx and ESBL activity identified by qRefCx AST).

23

The FilmArray Pheumonia Parel carbapenen reporting was also compared to the standard phenotypic carbapenen susceptibility testing methods performed in conjunction with current CLSI guidelines, standard phenotypic erapenen susceptibility is not reported for A. calcoaceticus-baunamii complex, therefore carbapenen susceptibility for this organism. Resistance or intermediate-resistance or meropenem or meropenem resistance for this analysis. The correlation between FilmArray Preumonia Panel resistance genes in a particular specimen as compared to the phenotypic AST results of isolates recovered from the stratified by each applicable associated organism in Table 15 through Table 18.

BAL
Applicable Bacteria ResultNIMPKPCNDMVIMOverall
(any carbapenem resistance
gene)
PPANPAPPANPAPPANPAPPANPAPPANPA
Overall
(any applicable bacteria Detected)126a0/17
(0%)109/109
(100%)3/17
(17.6%)109/109
(100%)0/17
(0%)109/109
(100%)0/17
(0%)109/109
(100%)3/17
(17.6%)
[6.2-41.0%]109/109
(100%)
[96.6-100%]
Acinetobacter calcoaceticus-baumannii complex0----------
Enterobacter cloacae complex90/0
(-)9/9
(100%)0/0
(-)9/9
(100%)0/0
(-)9/9
(100%)0/0
(-)9/9
(100%)0/0
(-)9/9
(100%)
Escherichia coli120/0
(-)12/12
(100%)0/0
(-)12/12
(100%)0/0
(-)12/12
(100%)0/0
(-)12/12
(100%)0/0
(-)12/12
(100%)
Klebsiella aerogenes70/0
(-)7/7
(100%)0/0
(-)7/7
(100%)0/0
(-)7/7
(100%)0/0
(-)7/7
(100%)0/0
(-)7/7
(100%)
Klebsiella oxytoca20/0
(-)2/2
(100%)0/0
(-)2/2
(100%)0/0
(-)2/2
(100%)0/0
(-)2/2
(100%)0/0
(-)2/2
(100%)
Klebsiella pneumoniae group140/0
(-)14/14
(100%)0/0
(-)14/14
(100%)0/0
(-)14/14
(100%)0/0
(-)14/14
(100%)0/0
(-)14/14
(100%)
Proteus spp.60/0
(-)6/6
(100%)0/0
(-)6/6
(100%)0/0
(-)6/6
(100%)0/0
(-)6/6
(100%)0/0
(-)6/6
(100%)
Pseudomonas aeruginosa420/12
(0%)30/30
(100%)0/12
(0%)30/30
(100%)0/12
(0%)30/30
(100%)0/12
(0%)30/30
(100%)0/12
(0%)30/30
(100%)
Serratia marcescens60/0
(-)6/6
(100%)0/0
(-)6/6
(100%)0/0
(-)6/6
(100%)0/0
(-)6/6
(100%)0/0
(-)6/6
(100%)
Polymicrobial specimens280/5
(0%)23/23
(100%)3/5
(60.0%)23/23
(100%)0/5
(0%)23/23
(100%)0/5
(0%)23/23
(100%)3/5b
(60.0%)23/23
(100%)

Table 15. IMP, KPC, NDM, and VIM Performance Table (comparison to phenotypic AST methods for BAL specimens)

a The isolate recovered from one specimen (P. aeruginosa) did not yield valid AST results on the VITEK instrument

"Of the specines that wee species: A. calculario is and K. premovive group dectably First.exp. (A. catesteach bumanni isolated by ReCt., compresence esstance dentified by aReflect in the internet in the internet in the experience in close complex and P , arregions detected by Film, and industrial in the E. cloace issue by aReC. AST, and RPC isential in the E. cloace sop. and P. eengines descted by Film.tray (P. eengines instant identified by gleC. AST, and Kentified in the inder by PCR (see speciment that ween to concertainment) i one presiment in and experience by Fin.html (P. eenginns issued by alled by allet in the by allet in the issue by PCR sont included in the issuate by PCR sont complex and K. arrygere dected by Film.htm (E. coace isolated by qRefCx, carbapenem resistance identified by qRefCx AST and KPC not identified in the isolate by PCVseq).

24

Sputum
Applicable Bacteria ResultNIMPKPCNDMVIMOverall
(any carbapenem resistance
gene)
PPANPAPPANPAPPANPAPPANPAPPANPA
Overall
(any applicable bacteria Detected)229a0/35
(0%)194/194
(100%)6/35
(17.1%)193/194
(99.5%)0/35
(0%)194/194
(100%)1/35
(2.9%)194/194
(100%)6/35b
(17.1%)
[8.1-32.7%]193/194
(99.5%)
[97.1-
99.9%]
Acinetobacter calcoaceticus-baumannii complex5c0/2
(0%)3/3
(100%)0/2
(0%)3/3
(100%)0/2
(0%)3/3
(100%)0/2
(0%)3/3
(100%)0/2
(0%)3/3
(100%)
Enterobacter cloacae complex7d0/1
(0%)6/6
(100%)0/1
(0%)6/6
(100%)0/1
(0%)6/6
(100%)0/1
(0%)6/6
(100%)0/1
(0%)6/6
(100%)
Escherichia coli100/0
(-)10/10
(100%)0/0
(-)10/10
(100%)0/0
(-)10/10
(100%)0/0
(-)10/10
(100%)0/0
(-)10/10
(100%)
Klebsiella aerogenes30/0
(-)3/3
(100%)0/0
(-)3/3
(100%)0/0
(-)3/3
(100%)0/0
(-)3/3
(100%)0/0
(-)3/3
(100%)
Klebsiella oxytoca40/0
(-)4/4
(100%)0/0
(-)4/4
(100%)0/0
(-)4/4
(100%)0/0
(-)4/4
(100%)0/0
(-)4/4
(100%)
Klebsiella pneumoniae group210/2
(0%)19/19
(100%)1/2
(50.0%)19/19
(100%)0/2
(0%)19/19
(100%)0/2
(0%)19/19
(100%)1/2
(50.0%)19/19
(100%)
Proteus spp.60/0
(-)6/6
(100%)0/0
(-)6/6
(100%)0/0
(-)6/6
(100%)0/0
(-)6/6
(100%)0/0
(-)6/6
(100%)
Pseudomonas aeruginosa670/16
(0%)51/51
(100%)0/16
(0%)51/51
(100%)0/16
(0%)51/51
(100%)0/16
(0%)51/51
(100%)0/16
(0%)51/51
(100%)
Serratia marcescens14e0/1
(0%)13/13
(100%)1/1
(100%)13/13
(100%)0/1
(0%)13/13
(100%)0/1
(0%)13/13
(100%)1/1
(100%)13/13
(100%)
Polymicrobial specimens920/13
(0%)79/79
(100%)4/13
(30.8%)78/79
(98.7%)0/13
(0%)79/79
(100%)1/13
(7.7%)79/79
(100%)4/13f
(30.8%)78/79
(98.7%)

Table 16. IMP, KPC, NDM, and VIM Performance Table (comparison to phenotypic AST methods for sputum specimens)

a The isolate recovered from one specimen (P. aeruginosa) did not yield valid AST results on the VITEK instrument

b One specimen had presence of dual AMR genes (KPC and VIM)

& An A. calcoaceticus-baumannii isolate was not recovered by qRefCx for one specimen

d An E. cloacae isolate was not recovered by qRefCx for one specimen

& A S. marcescens isolate was not recovered by qRefCx for one specimen

"Of the for specines that vers oncenter A rakoaeticus-camanine complex. K. acregores, K. penginse detected on Film-tray (A. calcacetivs-samazani, K. neumone, and P. acrument is and of the C. and promote in all the sistem of the K. premoment what by PCS. sand TM identified in the P. acoginos inde by PCR (sq); cne speciment . calcaceticas-samani . comp. Priess sp., and P. centraliano (K. permania: and P. ecoginos isolad by ReC's. congoner resistant itentified in the K. peamnine in the K. pad KPC dentified in the K. memoriae islate by P.C.Sea), cre specimer E. coli and K. meammine ervon descted by Timetry and solded by R (currences resistentified in the K. premiment in the solate of PC (entified in the solate of PCReag) one species and S. marces cased by Film Army and soluted by ReC. (carbopener resilance in the S. marces in the solate by Releag. Of the notate by P.C. (sept. Of the new speciment were not consocious were and consocious were and ountplex and Protected by Film-tray and issuate by decided in the in the A. calconcerns womannii solar by quell. AST and KPC in dentified in ether issue in ether issue in eth specified in the learner and P . excurines included by the C., carteporten resistance identified by (ReC. AST, and KP in the solate by P. C. AST, and KP. not icentified in th E. coll and P. acruman descted by Filmlery and issisted in the P. ecusions issues issues by Refl. AST and KP ont identified in einer in einer in einer in einer in einer in ei premaine government detected by FinArray radi solated by aReC. (carapean resistence identified in the K. newmaniae isolae by PC Norther in ether isolar by PC Nor is of dentif speciment Process on 1 . actualized by Film hand by ReC. (carbopener ressione dentified in the P. acquinse in added by dReck is and K.C. AST and Kerisolar in ether islar by P speciner P. acception and S. market by dief. (1 carberen resistance identified in the S. marcesens isolately of Reflix AST and KPC on i KPC ont i election in effective in eff one specime P. acrypinss and S. marces and by Pinters (P. acquines issues ilentified by aRellined by aReliev in the inder the investment of calcasedenas-bannanii comber, E. col. K. preumos detected by Film.traw (E. oli ad P. actupener resistence included in the P. acropeners resistence in the P. acupinses inder b

25

oller C. AST, and KP in isher issiate by PCReag) one speciment. A. calosetics-bannami company, Protess spp., P. acruginos, and S. marces cased by Film-low sprand P. actively isolated by qRefCx; carbapenen resistance in the P. aeruginosa isolate by qRefCx AST and KPC not identified in either isolate by PCR/seq).

BAL
Applicable Bacteria ResultPositive Percent AgreementNegative Percent Agreement
TP/(TP + FN)%95%CITN/(TN + FP)%95%CI
Overall
(any applicable bacteria Detected)0/20-77/7710095.2-100%
Enterobacter cloacae complex0/10-9/910070.1-100%
Escherichia coli0/0--13/1310077.2-100%
Klebsiella aerogenes0/0--7/710064.6-100%
Klebsiella oxytoca0/0--3/310043.9-100%
Klebsiella pneumoniae group0/0--15/1510079.6-100%
Proteus spp.0/0--6/610061.0-100%
Serratia marcescens0/0--8/810067.6-100%
Polymicrobial specimens0/1a0-16/1610080.6-100%

Table 17. OXA-48-like Performance Table (comparison to phenotypic AST methods for BAL specimens)

2 E. cloace complex and K. aerogenes detected by FilmArray (E. cloacae complex isolated by qRefCx and carbapenem resistance identified by qRefCx AST)

Sputum
Applicable Bacteria ResultPositive Percent AgreementNegative Percent Agreement
TP/(TP + FN)%95%CITN/(TN + FP)%95%CI
Overall
(any applicable bacteria Detected)0/100-121/12110096.9-100%
Enterobacter cloacae complex0/10-8/810067.6-100%
Escherichia coli0/0--17/1710081.6-100%
Klebsiella aerogenes0/0--4/410051.0-100%
Klebsiella oxytoca0/0--5/510056.6-100%
Klebsiella pneumoniae group0/30-22/2210085.1-100%
Proteus spp.0/0--9/910070.1-100%
Serratia marcescens0/30-22/2210085.1-100%
Polymicrobial specimens0/3a0-34/3410089.8-100%

Table 18. OXA-48-like Performance Table (comparison to phenotypic AST methods for sputum specimens)

² One spesimer E. ool and K. preumoniae group detated by qRefC: (carbpenen resistance in the K. preumonies group isolate by qRefC: AST); on especinen K. aerogenes and K. preumonie group isotaled by (ReCx and carbapenen resistance identified by (ReC. AST), one speiment. In neumoriae group and Proteus spp. detected by FilmArray (K. pneumoniae group isolated by qRefCx and carbapenem resistance identified by qRefCx AST)

26

The FilmArray Pneumonia Panel mec.A/C and MREJ reporting was also compared to the standard phonotypic cefoxitin susceptibility testing methods performed in conjunction with qRefCx. The correlation between FilmArray Pheumonia Panel reporting of mecAC and MREJ in a particular specimen as compared to the phenotypic AST results of isolates recovered from the same specimen is shown in Error! Reference source not found. and Table 19 and Table 20Error! Reference source not found..

BAL
S. aureus
mecA/C and MREJqRefCx: S. aureus
qRefCx Phenotypic AST: cefoxitin susceptibility
Org+/ Res+Org+/ Res-Org -Total
FilmArray
ResultOrg+/ Res+1832546
Org+/ Res-1244570
Org -01729730
Total1928799846
PerformanceAgreement%95%CI
Org+/ Res+18/1994.7%75.4-99.1%
Org+/ Res-24/2885.7%68.5-94.3%
Org -729/79991.2%89.1-93.0%
InterpretationPPANPAPrevalence
MRSA18/19
(94.7%)799/827
(96.6%)46/846
(5.4%)
MSSA24/28
(85.7%)772/818
(94.4%)70/846
(8.3%)
S. aureus46/47
(97.9%)729/799
(91.2%)116/846
(13.7%)

Table 19. mecA/C and MREJ 3x3 Performance Table (qRefCx & phenotypic AST on cultured isolate(s) from BAL specimens)

Table 20. mecA/C and MREJ 3x3 Performance Table (qRefCx & phenotypic AST on cultured isolate(s) from sputum specimens)

Sputum
FilmArray
ResultS. aureus
mecA/C and MREJqRefCx: S. aureus
qRefCx Phenotypic AST: cefoxitin susceptibility
Org+/ Res+Org+ / Res-Org -Total
Org+/ Res+59345107
Org+ / Res-1484897
Org -01631632
Total6052724836
PerformanceAgreement%95%CI
Org+ / Res+59/6098.3%91.1-99.7%
Org+ / Res-48/5292.3%81.8-97.0%
Org -631/72487.2%84.5-89.4%
InterpretationPPANPAPrevalence
MRSA59/60
(98.3%)728/776
(93.8%)107/836
(12.8%)
MSSA48/52
(92.3%)735/784
(93.8%)97/836
(11.6%)
S. aureus111/112
(99.1%)631/724
(87.2%)204/836
(24.4%)

27

The FilmArray Pneumonia Panel bin performance compared to the quantitative molecular assay (qMol) comparator is shown for BAL (Table 21) and sputum (Table 22). The qMol values are broken into onelog ranges correlating to the reported semi-quantitative FilmArray Pneumonia Panel bins. The relationship between qMol quantitative bins in copies/mL and traditional culture quantification in CFU/mL is unknown.

BAL
qMol Binned Valuesa
(copies/mL)ND to
Streptococcus pyogenes7/710064.6-100%8/810067.6-100%
Antimicrobial Resistance Genes
CTX-M1/1100-12/1210075.8-100%
Viruses
Influenza A2/210034.2-100%0/10-

Testing of Contrived Specimens

A prospective clinical evaluation of the FilmArray Pneumonia Panel was performed during the 2016-2017 respiratory infection season at several geographically diverse clinical laboratories. Over 1600 specimens were analyzed from subjects whose specimens were submitted for microbial evaluation of lower respiratory tract pathogens. In the prospective study, some analytes were of insufficient prevalence to adequately demonstrate system performance and additional archived, preselected positive specimens containing rare analytes were also tested. Several analytes were so rare that both prospective and archived testing efforts were insufficient to demonstrate system performance. In this study, contrived clinical specimens were created to evaluate the sensitivity and specificity of the FilmArray Pneumonia Panel assays for these rare analytes (Table 39Error! Reference source not found.).

36

Matrix
AnalyteBALSputum
Bacteria
Acinetobacter calcoaceticus-baumannii complexX
Klebsiella aerogenesXX
Klebsiella oxytocaX
Proteus spp.X
Serratia marcescensX
Streptococcus pyogenesXX
Atypical Bacteria
Chlamydia pneumoniaeXX
Legionella pneumophilaXX
Mycoplasma pneumoniaeXX
Viruses
AdenovirusXX
Human MetapneumovirusXX
Influenza AXX
Influenza BXX
Antibiotic Resistance Markers
CTX-MXX
IMPXX
KPCXX
NDMXX
OXA-48-likeXX
VIMXX

Table 39. Contrived Clinical Specimen Analytes

Contrived specimens (N=1125) were spiked using residual clinical samples that were pre-screened with the FilmArray Pneumonia Panel and found to be negative for the analytes of interest. Specimens were spiked with a variety of different isolates/strains for each organism at concentrations spanning observed ranges in clinical specimens. Different isolates of organisms were used in analytical testing when possible. Samples positive for one analyte served as negatives for other analytes.

For the majority of analytes reported qualitatively, at least 25 of the contrived positive specimens had analyte concentrations at 2 × the limit of detection (LoD), while the remaining specimens were tested at additional concentrations that spanned clinically observed ranges. The "clinically observed range" was based on data from previous FilmArray Pneumonia Panel positive test results (e.g. observations from the prospective or archived studies). If a clinically observed range could not be determined for a particular analyte, specimens were spiked at various factors of LoD. If the stock concentration of organism did not allow for spiking at the highest level, the highest achievable level was used. For bacteria reported with binned values, specimens were spiked at various concentrations starting just below and then spanning the reported levels (i.e. 10^3 to ≥10^7 copies per milliliter (mL)).

Specimens were prepared and randomized at BioFire such that the analyte status of each contrived specimen was unknown to the users performing the testing. BAL and sputum specimens were analyzed separately; however, the preparation and testing for both matrices were identical. Contrived specimens

37

were frozen, then distributed to prospective study sites and tested according to the prospective clinical study protocol alongside clinical (non-contrived) specimens.

The positive percent agreement (PPA) and negative percent (NPA) for the FilmArray Pneumonia Panel assays were determined using standard binomial sampling statistics. In this study, a success was defined as agreement between the known composition of the contrived specimen and the FilmArray Pneumonia Panel result; i.e., a positive FilmArray Pneumonia Panel result for spiked samples (True Positive, TP) and a negative FilmArray Pneumonia Panel result for un-spiked samples (True Negative, TN).

The results of the 1125 specimens tested in this study are summarized in Error! Reference source not found. for BAL and Error! Reference source not found. for sputum below.

The majority of analytes in both specimen types met the performance goals of 90% PPA with an 80% lower bound of the 95% Cl and 98% NPA with a 95% lower bound of the 95% CI. The exceptions being influenza A spiked into BAL and Klebsiella aerogenes spiked into BAL and sputum. Influenza A spiked in BAL demonstrated 86% PPA in part due to two missed detections at 0.2 × LoD and two additional missed detections at 2 × LoD from a strain that may have been under-quantified. However, FilmArray performance goals for influenza A in BAL were achieved in the archived study. Klebsiella aerogenes spiked in both sample types demonstrated 85.5% PPA in part due to five missed detections in BAL and four missed detections in sputum at a range of concentrations from a Klebsiella aerogenes strain that demonstrated poor reactivity with the FilmArray Pneumonia Panel.

Samples spiked with bacterial analytes just below the 10% copies/mL binned value (i.e., near 1.00E+03), and samples spiked with other analytes below their LoD (i.e., near 0.2 × LoD), produced the expected unreliable detection.

Sensitivity/PPASpecificity/NPA
AnalyteTP/(TP + FN)%95% CITN/(TN + FP)%95% CI
Bacteria
Acinetobacter calcoaceticus-baumannii complex47/5094.083.8-97.9%598/59810099.4-100%
Klebsiella aerogenesa47/5585.573.8-92.4%592/59499.798.8-99.9%
Klebsiella oxytoca46/5092.081.2-96.8%604/60410099.4-100%
Proteus spp.48/5096.086.5-98.9%603/60310099.4-100%
Serratia marcescens49/5098.089.5-99.6%604/60410099.4-100%
Streptococcus pyogenes49/5098.089.5-99.6%597/59710099.4-100%
Atypical Bacteria
Chlamydia pneumoniae47/5094.083.8-97.9%604/60410099.4-100%
Legionella pneumophila50/5010092.9-100%599/59910099.4-100%
Mycoplasma pneumoniae48/5096.086.5-98.9%603/60499.899.1-100%
Viruses
Adenovirus50/5394.384.6-98.1%568/56999.899.0-100%
Human Metapneumovirus50/5010092.9-100%597/59899.899.1-100%
Influenza Ab43/5086.073.8-93.0%585/58510099.3-100%
Influenza Bc47/5094.083.8-97.9%588/58999.899.0-100%
Antibiotic Resistance Markers

Table 40. FilmArray Pneumonia Panel Performance of Contrived BAL Specimens

38

Sensitivity/PPASpecificity/NPA
AnalyteTP/(TP + FN)%95% CITN/(TN + FP)%95% CI
CTX-M130/13010097.1-100%323/32499.798.3-100%
IMP45/4510092.1-100%412/41210099.1-100%
KPC53/5310093.2-100%400/40010099.1-100%
NDM53/5310093.2-100%404/40410099.1-100%
OXA-48-like53/5310093.2-100%307/30710098.8-100%
VIM58/5810093.8-100%399/39910099.0-100%
  • Five FN specimens were spiked with an K. aerogenes) strain (ATCC 2975) that demonstrated poor reactivity with the FilmArray Pneumonia Panel (see Error! Reference source not found.).

b Two FN specimens were spiked with an influenza A strain that may have been under-quantified

6 Three FN specimens were spiked with an influenza B strain that may have been under-quantified

Table 41. FilmArray Pneumonia Panel Performance of Contrived Sputum Specimens

Sensitivity/PPASpecificity/NPA
AnalyteTP/(TP + FN)%TN/(TN + FP)%
Bacteria
Klebsiella aerogenesa47/5585.5513/513100
Streptococcus pyogenes48/5096.0516/516100
Atypical Bacteria
Chlamydia pneumoniae49/5098.0521/521100
Legionella pneumophila50/50100521/521100
Mycoplasma pneumoniae48/5096.0521/521100
Viruses
Adenovirus50/5296.2494/494100
Human Metapneumovirus51/51100520/520100
Influenza Ab47/5094.0517/52199.2
Influenza Bc48/5194.1516/51799.8
Antibiotic Resistance Markers
CTX-M121/12299.2289/29099.7
IMP43/4497.7381/381100
KPC54/54100360/36199.7
NDM53/53100372/372100
OXA-48-like51/51100232/232100
VIM56/56100369/369100

" Four FN specimens were spiked with an K. aerogenes) strain (ATCC 2975) that demonstrated poor reactivity with the FilmAray Pneumonia Panel (see Error! Reference source not found.).

b Two FN specimens were spiked with an influenza A strain that may have been under-quantified

6 Two FN specimens were spiked with an influenza B strain that may have been under-quantified

Testing of Polymicrobial Contrived Specimens

Additionally, two sets of individual BAL (N=60) and sputum (N=60) specimens were multi-spiked with randomized low, medium and high relative concentrations of either A. baumannii, E. cloacae, and E. coli or K. oxytoca, P. mirabilis, and S. marcescens. As shown in Table 43, the majority of the spiked organisms were reported at the expected relative low, medium, or high bin level by the

39

FilmArray Pneumonia Panel. In four BAL specimens, E. cloacae was intended to be spiked at a medium level but was reported in a in a high (≥10^7) bin. Also, one specimen spiked with a high level of P. mirabilis was not detected (i.e. a false negative result for P. mirabilis).

Organism Spiked Into Sputum
Spike LevelFilmArray ResultTotal
LowMediumHigh
Low
(104 copies/mL)600060
Medium
(105.5 copies/mL)060060
High
(107 copies/mL)006060

Table 42. Polymicrobial Sputum Specimen Results

Table 43. Polymicrobial BAL Specimen Results

Organism Spiked Into BAL
FilmArray Result
Spike LevelLowMediumHighTotal
Low
(104 copies/mL)600060
Medium
(105.5 copies/mL)0564a60
High
(107 copies/mL)005959b

ªQuantified at the bin boundary and reported as>=10^7

bOne false negative result

Selected Analytical Studies

Limit of Detection

A limit of detection (LoD) was established for atypical bacteria and viruses detected by the FilmArray Pneumonia Panel. LoD was estimated by testing dilutions of contrived BAL or sputum samples containing known concentrations of organisms. Confirmation of LoD was achieved by testing at least 20 replicates per samples type on FilmArray 2.0 and FilmArray Torch systems (60 replicates total per sample type). LoD concentration was confirmed when the analyte was detected in at least 95% of the replicates tested.

The confirmed LoD for each atypical bacterium or virus (including a LoD for more than one isolate of the more genetically diverse viruses) is listed in Table 44Error! Reference source not found.. LoD concentration is based on quantification of each culture in viable units (TCID50/mL or CFU/mL) and a

40

corresponding molecular LoD concentration (DNA or RNA copies/mL) is provided based on quantitative real-time or digital PCR.

Table 44. Summary of Limit of Detection (LoD) for FilmArray Pneumonia Panel Atypical Bacteria and Viruses
AnalyteIsolate
Strain/Serotype/Source IDLoD Concentrationa
Viable UnitsMolecular (DNA or RNA)
Atypical Bacteria
Chlamydia pneumoniaeTW183
ATCC VR-22825.0E-01 TCID50/mLb3.3E+02 copies/mLb
Legionella pneumophilaPhiladelphia-1
ATCC 331525.0E+02 CFU/mL1.6E+03 copies/mL
Mycoplasma pneumoniaeM129
Zeptometrix 08015797.5E+01 TCID50/mLb3.5E+03 copies/mLb
Viruses
AdenovirusSpecies A (A18)
ATCC VR-195.0E+01 TCID50/mL9.2E+03 copies/mL
Species B (B3)
Zeptometrix 0810062CF1.0E+00 TCID50/mL1.8E+03 copies/mL
Species C (C2)
ATCC VR-8465.0E+00 TCID50/mL7.5E+03 copies/mL
Species D (D37)
Zeptometrix 0810119CF2.5E-01 TCID50/mLb2.9E+03 copies/mLb
Species E (E4)
Zeptometrix 0810070CF1.0E-01 TCID50/mLb3.5E+04 copies/mLb
Species F (F41)
ATCC VR-9305.0E+00 TCID50/mL5.5E+03 copies/mL
229E
ATCC VR-7405.0E-01 TCID50/mL8.1E+01 copies/mL
CoronavirusHKU1
Clinical Specimend-1.0E+04 copies/mL
NL63
BEI NR-4702.5E+00 TCID50/mLc5.4E+02 copies/mLc
OC43
ATCC VR-7595.0E+02 TCID50/mLc9.3E+03 copies/mLc
Human Metapneumovirus16 Type A1
Zeptometrix 0810161CF5.0E+01 TCID50/mL5.9E+03 copies/mL
Human
Rhinovirus/
EnterovirusRhinovirus
Type 1A
Zeptometrix 810012CFN1.5E+01 TCID50/mLb6.6E+03 copies/mLb
Echovirus 6
Zeptometrix 0810076CF1.0E+02 TCID50/mL5.7E+02 copies/mL
Influenza AH1N1pdm09 A/SwineNY/03/09
Zeptometrix 0810249CF2.5E+00 TCID50/mLc1.7E+03 copies/mLc
H3N2
A/Port Chalmers/1/73
ATCC VR-8101.0E+00 TCID50/mLb2.1E+02 copies/mLb
Influenza BB/FL/04/06
Zeptometrix 0810255CF5.0E+00 TCID50/mLc4.2E+02 copies/mLc
Parainfluenza VirusType 1
Zeptometrix 0810014CF2.5E+01 TCID50/mL5.2E+03 copies/mL
Type 2
Zeptometrix 0810015CF2.5E+01 TCID50/mLc1.5E+03 copies/mLc
Type 3
Zeptometrix 0810016CF2.5E+01 TCID50/mLc3.8E+02 copies/mLc
Type 4A
Zeptometrix 0810060CF2.5E+02 TCID50/mL8.1E+03 copies/mL
Respiratory Syncytial VirusType A
Zeptometrix 0810040ACF1.0E+00 TCID50/mL4.3E+02 copies/mL

Table 44. Summary of Limit of Detection (LoD) for FilmArray Pneumonia Panel Atypical Bacteria and Viruses

3 The listed concentration was confirmed with ≥95% detection on each FilmArray system in artificial BAL (aBAL) and/or sputum.

b LoD confirmation (≥95% detection) was achieved at a 2 to 5-fold lower concentration in aBAL.

· LoD confirmation (≥95% detection) was achieved at a 2 to 5-fold lower concentration in sputum.

41

d No cultured isolates of Coronavirus HKU1 were available for testing.

Note: LoD concentrations of the cultured viruses and the obligate intracellular atypical bacteria (C. pneumoniae and M. pneumoniae) are provided in units of TCID50 (50% Tissue Culture Infectious Dose). TCIDso is an indirect measure of viral or bacterial concentration based on infectivity and cvtotoxicity and will therefore vary considerably depending on technique and methodology (including cell type, culture media and conditions, cytotoxicity of the virus, etc.). It is not appropriate to make determinations on relative sensitivity of different molecular assays for detection of viruses and bacteria based on LoD values measured in TCID50/mL. Concentrations are also presented in copies/mL based upon independent quantitative PCR assays (qPCR) or digital PCR. Note that the accuracy of qPCR assays may also be affected by assay conditions, the standard reference material, and sequence variance between strains.

No assay-specific LoD concentrations were determined for the bacterial analytes. For bacteria, the FilmArray Pneumonia Panel reports a Detected result when the estimated bacterial nucleic acid abundance is ≥10^3.5 copies/mL, and the panel reports a Not Detected result if there is no amplification or the estimated bacterial nucleic acid abundance is 0.95) and estimates of nucleic acid abundance and corresponding bin results were determined to be accurate within 0.5-log10 copies/mL when compared to a copies/mL input concentration determined by digital PCR.No assay-specific LoD concentrations were determined for the antimicrobial resistance (AMR) gene assays. AMR genes are reported as Detected when an applicable bacterium is Detected and the assay for the AMR gene is positive AMR gene assay results were observed in ≥95% of 90 replicates when the applicable bacterium was tested at a concentration ≥10^3.5 copies/mL in the precision evaluation (see Precision (Reproducibility) below).

Inclusivitv

Analytical reactivity of FilmArray Pneumonia Panel assays was evaluated via a combination of empirical (wet) testing and in silico analysis of sequences available in public databases. Testing was performed on a collection of more than 350 genetically diverse viruses, bacteria, and antimicrobial resistance genes. The tested isolates represent relevant species, strains, serotypes, or genotypes as well as temporal and geographic diversity for each of the panel analytes. Each isolate was tested in triplicate at concentrations near LoD or the lowest reportable level for the analyte. In silico analysis of sequence data was also used to make predictions of assay reactivity for less common strains or serotypes and AMR gene types that were not tested but that may be detected by the FilmArray Pneumonia Panel assays.

Atypical bacteria and viruses were tested and detected at concentrations within 3× LoD (Table 46Error! Reference source not found. - Table 56Error! Reference source not found.). Bacteria were tested at a concentration of 1.0E+04 copies/mL (based on digital PCR of a single-copy gene in the bacterial chromosome) and the majority of isolates (94.4%) were detected with the expected bin result (Table 57Error! Reference source not found. - Table 71Error! Reference source not found.) and when the bacterium was detected, the appropriate AMR gene(s) were also detected (Table 72Error! Reference source not found. - Table 79Error! Reference source not found.).

Limitations on assay reactivity (based on wet testing observations) with specific viral and bacterial isolates or sequences and AMR gene types or sequences are noted in Table 45Error! Reference source not found.. Most limitations are associated with single-base sequence variants under one or more assay primers. Additional predicted limitations on reativity based on in-silico sequence analysis are provided in the footnotes and analyte-specific tables below.

42

Note: FilmArray Pneumonia Panel Influenza B assays are predicted to react with attenuated viruses used in vaccines.

| Limitation | Observed
Result | Analyte | Strain/Isolate/Variant |
|------------|---------------------------------------------------------------------|---------------------------------------------------|----------------------------------------------------------------------|
| Minor | Detected
may be under-reported by
one bin (≤10-fold) | Enterobacter cloacae complex | Enterobacter hormaechei
(ATCC 49162)a |
| | | Klebsiella pneumoniae group | Klebsiella quasipneumoniae subsp.
quasipneumoniae
(DSM 28211)b |
| | | Moraxella catarrhalis | Moraxella catarrhalis
(ATCC 23246)c |
| | | Streptococcus pyogenes | Streptococcus pyogenes
(ATCC 19615) |
| Major | Detected
may be under-reported by
two or more bins (>10-fold) | Enterobacter cloacae complex | Enterobacter asburiae
(ATCC 35953, 35954, 35955, and 35957)d |
| | | Klebsiella aerogenes | Klebsiella (Enterobacter) aerogenes
(ATCC 29751)e |
| | | mecA/C and MREJ | MREJ type xv |
| | Not Detected | Acinetobacter calcoaceticus-
baumannii complex | Acinetobacter nosocomialis
(ATCC 700472)f |
| | | Pseudomonas aeruginosa | Pseudomonas aeruginosa
(ATCC 25619)g |
| | | mecA/C and MREJ | MREJ type xviii
MREJ type xix |

Table 45. Limitations on Analytical Reactivity of FilmArray Pneumonia Panel Assays Observed in Testing

3 Minor limitation observed for this isolate due to sequence variation predicted for two E. hormachei sequences (3.0%) from public databases.

b Minor limitation observed for this isolate due to sequence variant under a primer. Similar limitation predicted for one K. quasipneumonice sequence (20.0%) from public databases. Additional minor or major limitation predicted for four K. pneumoniae sequences. (0.3%) from public databases.

"Minor limitation observed for this isolate. Additional mintation predicted for two M. catarrhalis sequences (3.3%) from public databass.

4 Major limitation observed or predicted for these isonance variance under primer. Similar limitation predicted for five E. ashuriae squences (10.9%), eight E. cloacae sequences (2.2%), and two E. ludwigii sequences (16.7%) from public databases.

" Major limitation observed for this isolate duer primers. Similar limitation predicted for six K. aerogeners sequences (4.3%) from public databases

f ATCC 700472 could not be confirmed as A. nosocomialis by sequence and may be a non-Acinetobacticas-baumannii complex species that has been mis-identified.

8 Major limitation observed for this solate due to sequence variant unitation predicted for one P. ceriginosa sequence (0.7%) from public databases.

Table 46. Adenovirus Isolates Tested and Detected

OrganismSpeciesSerotype aSource [Strain/Location/Year]Test ConcentrationResult
(copies/mL)xLoD
AdenovirusA18ATCC VR-19 [Washington D.C./1954]9.2E+031x
12ATCC VR-863 [Huie/Massachusetts]2.7E+043x
31Zeptometrix 0810073CF2.7E+043x
B3Zeptometrix 0810062CF1.8E+031x
7ATCC VR-7 [Gomen/California/1954]5.3E+033x
7AZeptometrix 0810021CF5.3E+033x
7d/d2Univ of Iowa Research Foundation [Iowa/2001]5.3E+033x
7hUniv of Iowa Research Foundation [Iowa/1999]5.3E+033x
11ATCC VR-12 [Slobitski/Massachusetts]5.3E+033x
14ATCC VR-15 [De Wit/Netherlands/1955]5.3E+033x
Adenovirus16ATCC VR-17 [CH.79/Saudi Arabia/1955]5.3E+033xAdenovirus
Detected
21ATCC VR-1833 [128/Saudi Arabia/1956]5.3E+033x
34ATCC VR-716 [Compton/1972]5.3E+033x
35ATCC VR-718 [Holden]5.3E+033x
50ATCC VR-1602 [Wan/Amsterdam/1988]5.3E+033x
AdenovirusC2ATCC VR-846 [Adenoid 6]7.5E+031x
1Zeptometrix 0810050CF2.3E+043x
5Zeptometrix 0810020CF2.3E+043x
6ATCC VR-6 [Tonsil 99]2.3E+043x
37Zeptometrix 08100119CF5.8E+021x
D8Zeptometrix 0810069CF1.7E+033x
Adenovirus20Zeptometrix 0810115CF1.7E+033x

43

OrganismSpeciesSerotype aSource [Strain/Location/Year]Test ConcentrationResult
(copies/mL)xLoD
E4Zeptometrix 0810070CF1.7E+041x
4ATCC VR-1572 [RI-67/Missouri/1952-1953]1.0E+040.6x
4aUniv of Iowa Research Foundation [S Carolina/2004]1.0E+040.6x
F41ATCC VR-930 [Tak 73-3544/ Netherlands/1973]5.5E+031x
40NCPV 0101141v1.6E+043x
40Zeptometrix 0810084CF1.6E+043x
41Zeptometrix 0810085CF1.6E+043x

4 In silico analysis of available sequences predicts the FilmArray Pneumonia Parel with Adenovirus B55, C57, all species D serotyes, and G52.

Table 47. Coronavirus Isolates Tested and Detected

OrganismTypeSource [Location/Year]Test ConcentrationResult
(copies/mL)xLoD
Coronavirus229EATCC VR-7408.1E+011xCoronavirus
Detected
229EZeptometrix 0810229CF2.4E+023x
229EClinical Specimen [Utah/2015]1.0E+041x
HKU1aClinical Specimen [Detroit/2010]3.0E+043x
CoronavirusHKU1aClinical Specimen [Utah/2015]3.0E+043x
HKU1aClinical Specimen [Utah/2015]3.0E+043x
HKU1aClinical Specimen [S Carolina/2010]3.0E+043x
NL63BEI NR-470b [Amsterdam/2003]2.7E+021x
CoronavirusNL63Zeptometrix 0810228CF8.0E+023x
OC43ATCC VR-759c4.6E+031x
OC43Zeptometrix 0810024CF1.4E+043x

ª No cultured isolates of Coronavirus HKUI were available for testing. Five clinical NPS specimens HKUI were collected from different regions of the US in 2010 and 2015, quantified molecularly, and tested.

  • Organism obtained through the NH Biodefense and Emerging Infections Research Resources Repository, NIAID, NH: Human Coronavirus NL63, NR-470.

€ Discontinued part #. See ATCC VR-1558.

Table 48. Human Metapneumovirus Isolates Tested and Detected

OrganismGenotypeSerotypeSource [Location/Year]Test ConcentrationResult
(TCID50/mL)xLoD
Human
MetapneumovirusA116Zeptometrix 0810161CF [Iowa10/2003]5.0E+011xHuman
Metapneumovirus
Detected
9Zeptometrix 0810160CF [Iowa3/2002]1.5E+023x
A220Zeptometrix 0810163CF [Iowa14/2003]1.5E+023x
27Zeptometrix 0810164CF [Iowa27/2004]1.5E+023x
B13Zeptometrix0810156CF [Peru2/2002]1.5E+023x
5Zeptometrix 0810158CF [Peru3/2003]1.5E+023x
B28Zeptometrix 0810159CF [Peru6/2003]1.5E+023x
4Zeptometrix 0810157CF [Peru1/2002]1.5E+023x
18Zeptometrix 0810162CF [Iowa18/2003]1.0E+023x

Table 49. Human Rhinovirus and Enterovirus Isolates Tested and Detected

| Species | Serotype | Source [Strain/Location/Year] | Test Concentration
(copies/mL) | xLoD | Result |
|-------------------|-------------------|----------------------------------------|-----------------------------------|------|----------------------|
| Human Rhinovirusa | | | | | |
| | 1 | Zeptometrix 0810012CFN [1A] | 2.2E+03 | 1x | |
| | 2 | ATCC VR-482 [HGP] | 1.7E+03 | 3x | |
| | 7 | ATCC VR-1601 [68-CV11] | 1.7E+03 | 3x | |
| | 16 | ATCC VR-283 [11757/Washington DC/1960] | 1.7E+03 | 3x | |
| A | 34 | ATCC VR-507b [137-3] | 1.7E+03 | 3x | |
| | 57 | ATCC VR-1600 [Ch47] | 1.7E+03 | 3x | Human
Rhinovirus/ |
| | 77 | ATCC VR-1187 [130-63] | 1.7E+03 | 3x | Rhinovirus/ |
| | 85 | ATCC VR-1195 [50-525-CV54] | 1.7E+03 | 3x | Enterovirus |
| | 3 | ATCC VR-483 [FEB] | 1.7E+03 | 3x | Detected |
| | 14 | ATCC VR-284 [1059/S Carolina/1959] | 1.7E+03 | 3x | |
| | 17 | ATCC VR-1663 [33342/N Carolina/1959] | 1.7E+03 | 3x | |
| B | 27 | ATCC VR-1137 [5870] | 1.7E+03 | 3x | |
| | 42 | ATCC VR-338 [56822] | 1.7E+03 | 3x | |
| | 83 | ATCC VR-1193 [Baylor 7] | 1.7E+03 | 3x | |
| Enterovirus | | | | | |
| A | Coxsackievirus 10 | ATCC VR-168 [NY/1950] | 1.7E+03 | 3x | Human Rhinovirus/ |
| | Enterovirus 71 | ATCC VR-1432 [H] | 1.7E+03 | 3x | Enterovirus |

44

SpeciesSerotypeSource [Strain/Location/Year]Test ConcentrationResult
(copies/mL)xLoD
BCoxsackievirus A9Zeptometrix 0810017CF1.7E+033xDetected
Coxsackievirus B3Zeptometrix 0810074CF1.7E+033x
Coxsackievirus B4Zeptometrix 0810075CF1.7E+033x
Echovirus 6Zeptometrix 0810076CF5.7E+021x
Echovirus 9Zeptometrix 0810077CF1.7E+033x
Echovirus 11Zeptometrix 0810023CF1.7E+033x
CCoxsackievirus A21ATCC VR-850 [Kuykendall/California/1952]1.7E+033x
Coxsackievirus A24ATCC VR-583 [DN-19/Texas/1963]1.7E+033x
DEnterovirus 68ATCC VR-1823 [US/MO/2014-18947]1.7E+033x

4 The concentration used for Human Rhinovirus isolate testing was based on 3× the Enterovins LoD concentration (5.7E+02 copies/mL).

b Discontinued part #; see ATCC VR-1365.

Table 50. Influenza A Isolates Tested and Detected

| Organism | Subtype | Source [Strain/Location/Year] | Test Concentration
(copies/mL) xLoD | | Result | |
|-------------|--------------------------------|----------------------------------------------------|---------------------------------------------------------------------------------------|---------|--------|-------------------------|
| Influenza A | H1N1 | Human | | | | |
| | | ATCC VR-219 [NWS/1933] | 3.1E+02 | 3x | | |
| | | ATCC VR-95 [PR/8/1934a] | 1.0E+03 | 1.5xa | | |
| | | ATCC VR-96 [Wiess/1943] | 3.1E+02 | 3x | | |
| | | ATCC VR-97 [FM/1/1947] | 3.1E+02 | 3x | | |
| | | ATCC VR-98 [Mal/302/1954] | 3.1E+02 | 3x | | |
| | | ATCC VR-546 [Denver/1/1957] | 3.1E+02 | 3x | | |
| | | Zeptometrix 0810036CF [New Caledonia/20/1999] | 3.1E+02 | 3x | | |
| | | Zeptometrix 0810036CFN [Solomon Islands/3/2006] | 3.1E+02 | 3x | | |
| | | Zeptometrix 0810244CF [Brisbane/59/2007] | 3.1E+02 | 3x | | |
| | | H1N2 | BEI NR-3478b [Kilbourne F63 A/NWS/1934 (HA) x
A/Rockefeller Institute/5/1957 (NA)] | 3.1E+02 | 3x | |
| | | | Zeptometrix 0810249CF [SwineNY/03/2009] | 6.6E+02 | 1x | |
| | | H1N1pdm09 | Zeptometrix 0810109CFJ [Canada/6294/2009] | 3.1E+02 | 3x | |
| | | | Zeptometrix 0810165CF [California/07/2009] | 3.1E+02 | 3x | |
| | | | Zeptometrix 0810166CF [Mexico/4108/2009] | 3.1E+02 | 3x | |
| | | | BEI NR-19823c [Netherlands/2629/2009] | 3.1E+02 | 3x | |
| | | | BEI NR-42938d [Georgia/F32551/2012] | 3.1E+02 | 3x | |
| | | | BEI NR-44345e [Hong Kong/H090-761-V1(0)/2009] | 1.0E+03 | 1.5xa | |
| | | H3N2 | ATCC VR-810 [Port Chalmers/1/1973] | 1.0E+02 | 1x | Influenza A
Detected |
| | | | ATCC VR-776 [Alice (live attenuated vaccine)] | 3.1E+02 | 3x | |
| | | | Zeptometrix 0810238CF [Texas /50/2012] | 3.1E+02 | 3x | |
| | ATCC VR-547 [Aichi/2/1968] | | 3.1E+02 | 3x | | |
| | ATCC VR-544 [Hong Kong/8/1968] | | 3.1E+02 | 3x | | |
| | | ATCC VR-822 [Victoria/3/1975] | 3.1E+02 | 3x | | |
| | | Zeptometrix 0810252CF [Wisconsin/67/2005] | 3.1E+02 | 3x | | |
| | | Zeptometrix 0810138CF [Brisbane/10/2007] | 3.1E+02 | 3x | | |
| | H3N2v | ODHL1 [Ohio/2012] | 3.1E+02 | 3x | | |
| | Avian | | | | | |
| | H2N2 | BEI NR-2775f [Japan/305/1957] | 3.1E+02 | 3x | | |
| | H2N3 | MRIGlobalg [Mallard/Alberta/79/2003] | 3.1E+02 | 3x | | |
| | H5N1 | MRIGlobalg [Chicken/Yunnan/1251/2003] | 3.1E+02 | 3x | | |
| | H5N2 | MRIGlobalg [Northern pintail/Washinton/40964/2014] | 3.1E+02 | 3x | | |
| | H5N3 | BEI NR-9682h [Duck/Singapore/645/97] | 3.1E+02 | 3x | | |
| | H5N8 | MRIGlobalg [Gyrfalcon/Washing-ton/41088-6/2014] | 3.1E+02 | 3x | | |
| | H7N7 | MRIGlobalg [Netherlands/219/2003] | 3.1E+02 | 3x | | |
| | H7N9 | MRIGlobalg [Anhui/01/2013] | 3.1E+02 | 3x | | |
| | H10N7 | BEI NR-2765i [Chicken/Germany/N/49] | 3.1E+02 | 3x | | |
| | Swine | | | | | |
| | H1N1 Swine | ATCC VR-333 [Swine/Iowa/15/1930] | 3.1E+02 | 3x | | |
| | | ATCC VR-99 [Swine/1976/1931] | 3.1E+02 | 3x | | |

ª 1.5x the LoD for Influenza A H1N1pdm09 Zeptometrix 0810109CFN [SwineNY/03/2009] (6.6E+02 copies/mL).

6 Genomic RNA obtained through BEI Resources NAID, NH: Kibourne F63: ANWS/1934 (HA) x ARockefeller Institute/5/197 (HIN2), Reassortant NWS-F, NR-9677.

6 Virus obtained through BEI Resources, NIAID, NIH: Influenza A Virus, A/Netherlands/2629/2009 (H1N1)pdm09, NR-19823

d Virus obtained through BEI Resources, NIAID, NIH: Influenza A Virus, A/Georgia/F32551/2012 (H1N1)pdm09, NR-42938.

° Virus obtained through BEI Resources, NIAID, NH: Influenza A Virus, A/Hong Kong/H090-761-V1(0) 2009 (H1N1) pdm09, NR-44345.

f Genomic RNA obtained through BEI Resources, NIAID, NIH: Genomic RNA from Influenza A Virus, A/Japan/305/1957 (H2N2), NR-2775.

8 Isolate provided and tested by MRI Global, Kansas City, MO.

BioFire Diagnostics 510(k)

FilmArray Pneumonia Panel

45

4 Genomic RNA obtained through BEI Resources, NIAID, NIH: Genomic RNA from Influenza A Vins, Alduck/Singapore/645/1997 (H5N3), Wild Type, NR-9682.

' Genomic RNA obtained through BEI Resources, NIAID, NH: Genomic RNA from Influenza A Virus, A/chicken/Germany/N/1949 (H/0N/J/NR-2765.

| Organism
Lineage | | [Strain/Location/Year], Source | Test Concentration
(copies/mL) | | xLoD | Result |
|---------------------|----------|----------------------------------------------|-----------------------------------|--|------|-------------------------|
| Influenza B | | ATCC VR-101 [Lee/1940] | 6.3E+02 | | 3x | Influenza B
Detected |
| | | ATCC VR-102 [Allen/1945] | 6.3E+02 | | 3x | |
| | | ATCC VR-103 [GL/1739/1954] | 6.3E+02 | | 3x | |
| | | ATCC VR-296 [1/Maryland/1959] | 6.3E+02 | | 3x | |
| | | ATCC VR-295 [2/Taiwan/1962] | 6.3E+02 | | 3x | |
| | | ATCC VR-786 [Brigit/Russia/1969] | 6.3E+02 | | 3x | |
| Influenza B | Victoria | ATCC VR-823 [5/Hong Kong/1972] | 6.3E+02 | | 3x | |
| | | Zeptometrix 0810258CF [2506/Malaysia/2004] | 6.3E+02 | | 3x | |
| | | CDC 2005743348 [1/Ohio/2005] | 6.3E+02 | | 3x | |
| | Yamagata | Zeptometrix 0810256CF [07/Florida/2004] | 6.3E+02 | | 3x | |
| | | Zeptometrix 0810255CF [04/Florida/2006] | 2.1E+02 | | 1x | |
| | | Zeptometrix 0810241CF [1/Wisconsin/2010] | 6.3E+02 | | 3x | |
| | | Zeptometrix 0810239CF [2/Massachusetts/2012] | 6.3E+02 | | 3x | |

Table 51. Influenza B Isolates Tested and Detected

Table 52. Parainfluenza Virus Isolates Tested and Detected

OrganismTypeSource [Strain/Location/Year]Test ConcentrationResult
(copies/mL)xLoD
Parainfluenza Virus1Zeptometrix 0810014CF5.2E+031xParainfluenza Virus Detected
1BEI NR-48680ª [FRA/29221106/2009]1.6E+043x
1ATCC VR-94 [C-35/Washington DC/1957]1.6E+043x
2Zeptometrix 0810015CF1.5E+031x
2ATCC VR-92 [Greer/Ohio/1955]8.9E+020.6x
3Zeptometrix 0810016CF1.9E+021x
3BEI NR-3233ᵇ [NIH 47885, Wash/47885/57]5.7E+023x
3ATCC VR-93 [C-243/Washington DC/1957]5.7E+023x
4Zeptometrix 0810060CF8.1E+031x
4ATCC VR-1378 [M-25/1958]2.4E+043x
4Zeptometrix 0810060BCF2.4E+043x
4ATCC VR-1377 [CH-19503/Washington DC/1962]2.4E+043x

ª Virus obtained through BEI Resources, NIAID, NH: Human Parainfluenza Virus 1, HPV 1/FRA/29221106/2009, NR-4868.

b Virus obtained through BEI Resources, NIAID, NIH: Human Parainfluenza Virus 3, NIH 47885, NR-3233.

Table 53. Respiratory Syncytial Virus Isolates Tested and Detected

| Organism | Type | Source [Strain/Location/Year] | Test Concentration
(copies/mL) | | xLoD | Result |
|--------------------------------|------|-----------------------------------------|-----------------------------------|--|------|--------------------------------------------|
| Respiratory
Syncytial Virus | A | Zeptometrix 0810040ACF [2006] | 4.3E+02 | | 1x | Respiratory
Syncytial Virus
Detected |
| | | ATCC VR-26 [Long/Maryland/1956] | 1.3E+03 | | 3x | |
| | | ATCC VR-1540 [A2/Melbourne/1961] | 1.3E+03 | | 3x | |
| | B | Zeptometrix 0810040CF [Ch-93 (18)-18] | 1.3E+03 | | 3x | |
| | | ATCC VR-1400 [WV/14617/1985] | 1.3E+03 | | 3x | |
| | | ATCC VR-955 [9320/Massachusetts/1977] | 1.3E+03 | | 3x | |
| | | ATCC VR-1580 [18537/Washington DC/1962] | 1.3E+03 | | 3x | |

Table 54. Chlamydia pneumoniae Isolates Tested and Detected

Test Concentration
OrganismSource [Strain](copies/mL)xLoDResult
Chlamydia
pneumoniaeATCC VR-2282 [TW-183/Taiwan/1965]6.7E+011xChlamydia pneumoniae Detected
ATCC VR-1310 [CWL-029]2.0E+023xChlamydia pneumoniae Detected
ATCC VR-1360 [CM-1/Georgia]2.0E+023xChlamydia pneumoniae Detected
ATCC 53592 [AR-39/Seattle/1983]2.0E+023xChlamydia pneumoniae Detected

Table 55. Legionella pneumophila Isolates Tested and Detected

Species/SubspeciesSerogroupSource [Strain]Test ConcentrationResult
(CFU/mL)xLoD
L. pneumophila1ATCC 33152 [Philadelphia-1]$5.0E+02$1xLegionella
pneumophila
Detected
L. pneumophila3ATCC 33155 [Bloomington-2]$1.5E+03$3x
L. pneumophila subsp. fraseri4ATCC 33156 [Los Angeles-1]$1.5E+03$3x
L. pneumophila subsp. fraseri5ATCC 33216 [Dallas 1E]$1.5E+03$3x
L. pneumophila subsp. pascullei5ATCC 33737 [U8W]$1.5E+03$3x
L. pneumophila subsp. pneumophila10ATCC 43283 [Leiden 1]$1.5E+03$3x

46

Species/SubspeciesSerogroupSource [Strain]Test ConcentrationResult
14ATCC 43703 [1169-MN-H]1.5E+033x
(CFU/mL)xLoD
Table 56. Mycoplasma pneumoniae Isolates Tested and Detected
OrganismTypeSource [Strain]Test ConcentrationResult
(copies/mL)xLoD
Mycoplasma
pneumoniae1Zeptometrix 0801579 [M129]1.2E+031xMycoplasma
pneumoniae
Detected
1ATCC 29342 [M129-B7]3.5E+033x
1ATCC 29085 [PI 1428]3.5E+033x
2ATCC 15531-TTR [FH strain of Eaton Agent [NCTC
10119]]3.5E+033x
unknownATCC 15492 [Mac]3.5E+033x
unknownATCC 15293 [M52]3.5E+033x
unknownATCC 15377 [Bru]3.5E+033x
unknownATCC 39505 [Mutant 22]3.5E+033x
unknownATCC 49894 [UTMB-10P]3.5E+033x

Table 57. Acinetobacter calcoaceticus-baumannii complex Isolates Tested and Detected

OrganismSource [Strain]Test concentration (copies/mL)Result
A. baumanniiATCC 9955 [6-561]1.0E+04Acinetobacter
calcoaceticus-
baumannii complex
Detected
A. baumanniiATCC 19606 [2208 Type strain]1.0E+04
A. baumanniiATCC 17961 [CDC 7788]1.0E+04
A. baumanniiAR-Bank #00331.0E+04
A. baumanniiGRE 11530641.0E+04
A. baumanniiGRE 10620811.0E+04
A. calcoaceticusATCC 514321.0E+04
ATCC 23055 [46]1.0E+04
ATCC 14987 [HO-1]1.0E+04
A. calcoaceticus subsp. anitratusATCC 15308 [NCTC 7844]1.0E+04
A. pittiiATCC 19004 [57.071.228]1.0E+04
A. nosocomialisaATCC 17903 [NCTC 8102]1.0E+04
A.seifertiiCCUG 347851.0E+04

a A. nosocomialis ATCC 700472 was not detected at any concentration. Sequencing suggests the isolate may be mis-identified.

Table 58. Enterobacter cloacae complex Isolates Tested and Detected

OrganismSource [Strain]Test concentration (copies/mL)Result
E. cloacaeATCC 49141 [AmMs 204]1.0E+04
E. cloacaeATCC BAA-1143 [Entb 55M]1.0E+04
E. cloacaeATCC BAA-2341 [1101152]1.0E+04
E. cloacaeAR-Bank #01541.0E+04
E. cloacaeNCTC 134641.0E+04
E. cloacae subsp. cloacaeATCC 13047 [Type Strain]1.0E+04Enterobacter
E. cloacae subsp. dissolvensATCC 23373Dª [ICPB ED105]1.0E+04cloacae complex
E. asburiaeATCC 35953b [CDC 1497-78 Type Strain]1.0E+06bDetected
ATCC 35957b [CDC 570-83]1.0E+06b
E. hormaecheiATCC 49162b [CDC 992-77]1.0E+05b
E. hormaecheiATCC BAA-20821.0E+04
E. kobeiCCUG 594101.0E+04
E. ludwigiiCCUG 230501.0E+04

ª Genomic DNA from E. cloacae subsp. dissolvens.

b See Table 45Error! Reference source not found. for limitation.

Table 59. Escherichia coli Isolates Tested and Detected

OrganismSource [Strain]Test concentration (copies/mL)Result
E. coliATCC 25922 [FDA strain Seattle 1946]1.0E+04Escherichia coli
Detected
ATCC 43888 [CDC B568-73]1.0E+04
AR-Bank #00611.0E+04
AR-Bank #00861.0E+04
AR-Bank #01371.0E+04
AR-Bank #01501.0E+04
AR-Bank #01621.0E+04
GRE 10620161.0E+04
GRE 12520081.0E+04
GRE 12520091.0E+04
GRE 12560181.0E+04
Zeptometrix 0801905 [Z136]1.0E+04
ATCC 29930 [WRAIR I virulent]1.0E+04

47

Table 60. Haemophilus influenzae Isolates Tested and Detected

OrganismSerotypeSource [Strain/Location/Year]Test concentration (copies/mL)Result
H. influenzaeType aATCC 9006 [AMC 36-A-3 [610, PCM 2436]]1.0E+04Haemophilus
influenzae Detected
Type bATCC 10211 [AMC 36-A-1 [572]], Biotype 11.0E+04
Type cATCC 49699 [C 9007]1.0E+04
Type dATCC 9008 [AMC 36-A-6 [611]]1.0E+04
Type eATCC 8142 [AMC 36-A-7 [595, NCTC 8472]]1.0E+04
Type fATCC 700223 [GA1264]1.0E+04
Biogroup
aegyptiusATCC 11116 [180-a [NCTC 8502]]1.0E+04
Non-typeableATCC 51907 [Rd [KW20]]1.0E+04

NOTE: The Hinfluenzae assay will not react with strains that do not carry the hpd gene108

Table 61. Klebsiella (Enterobacter) aerogenes Isolates Tested and Detected

OrganismSource [Strain]Test concentration (copies/mL)Result
K. aerogenesATCC 13048 [NCTC 10006]1.0E+04Klebsiella
aerogenes
Detected
AR-Bank #00621.0E+04
AR-Bank #00741.0E+04
AR-Bank #01611.0E+04
GRE 12540661.0E+04
ATCC 29751a [MULB-250]1.0E+07a

ªPreviously known as Enterobacter aerogenes.

b See Table 45Error! Reference source not found. for limitation.

Table 62. Klebsiella oxytoca Isolates Tested and Detected

OrganismSource [Strain]Test concentration (copies/mL)Result
K. oxytocaATCC 13182 [479-2 Type strain]1.0E+04Klebsiella oxytoca
Detected
ATCC 43086 [Pasco 201]1.0E+04
ATCC 49131 [AmMS 101]1.0E+04
ATCC 700324 [LBM 90.11.033]1.0E+04
ATCC 8724 [NRRL B-199]1.0E+04
AR-Bank #01471.0E+04
JMI 25231.0E+04
JMI 26611.0E+04
JMI 78181.0E+04
JMI 106781.0E+04
JMI 146111.0E+04
GRE 12540541.0E+04

Table 63. Klebsiella pneumoniae Isolates Tested and Detected

OrganismSource [Strain]Test concentration (copies/mL)Result
K. pneumoniaeATCC BAA-1705 [ART 2008133]1.0E+04Klebsiella
pneumoniae
Detected
AR-Bank #00681.0E+04
AR-Bank #00751.0E+04
AR-Bank #00761.0E+04
AR-Bank #00791.0E+04
AR-Bank #00801.0E+04
AR-Bank # 00971.0E+04
AR-Bank #01071.0E+04
AR-Bank #01531.0E+04
GRE 10620841.0E+04
GRE 13550301.0E+04
JMI 3281.0E+04
JMI 7661.0E+04
NCTC 134651.0E+04
Zeptometrix 08018861.0E+04
K. pneumoniae subsp. ozaenaeATCC 11296 [AMC 35-E-5]1.0E+04
K. pneumoniae subsp. pneumoniaeATCC 13883 [NCTC 9633]1.0E+04
K. pneumoniae subsp. rhinoscleromatisATCC 13884 [NCTC 5046]1.0E+04
K. quasipneumoniae subsp. quasipneumoniaeDSM 28211ª [01A030, SB11]1.0E+05a
K. quasipneumoniae subsp. simipneumoniaeDSM 28212 [07A044, SB30]1.0E+04
K. variicolaATCC BAA-830 [F2R9]1.0E+04

ª See Table 45Error! Reference source not found. for limitation.

Table 64. Moraxella catarrhalis Isolates Tested and Detected

| Organism | Source [Strain] | Test concentration
(copies/mL) | Result |
|----------------------------|-----------------|-----------------------------------|--------|
| BioFire Diagnostics 510(k) | | | |

48

| M. catarrhalis | ATCC 25238 [Ne 11] | 1.0E+04 | Moraxella catarrhalis
Detected |
|-----------------------|-------------------------|----------|------------------------------------------|
| | ATCC 25240 [N9] | 1.0E+04 | |
| | ATCC 8176 [20] | 1.0E+04 | |
| | ATCC 23246a [NCTC 4103] | 1.0E+05a | |
| | ATCC 49143 [Am MS 116] | 1.0E+04 | |

ª See Table 45Error! Reference source not found. for limitation.

Table 65. Proteus spp. Isolates Tested and Detected

| Organism | Source [Strain] | Test concentration
(copies/mL) | Result |
|---------------------|---------------------------------------|-----------------------------------|--------------------------|
| P. mirabilis | ATCC 29906 [1003] | 1.0E+04 | Proteus spp.
Detected |
| | ATCC 33583 [571101] | 1.0E+04 | |
| | ATCC 35659 [LRA 08 01 73] | 1.0E+04 | |
| | AR-Bank #0156 | 1.0E+04 | |
| | AR-Bank #0159 | 1.0E+04 | |
| | GRE 1254053 | 1.0E+04 | |
| P. hauseri | ATCC 13315 [NCTC 4175 Strain Lehmann] | 1.0E+04 | |
| | ATCC 700826 [CDC 1732-80] | 1.0E+04 | |
| P. penneri | ATCC 33519 [Type Strain CDC 1808-73] | 1.0E+04 | |
| | ATCC 35197 [CDC 1655-67] | 1.0E+04 | |
| P. vulgaris | ATCC 29905 | 1.0E+04 | |
| | ATCC 33420 | 1.0E+04 | |
| | ATCC 27973 [CDC 1787-64-SC1] | 1.0E+04 | |

Table 66. Pseudomonas aeruginosa Isolates Tested and Detected

OrganismSource [Strain]Test concentration (copies/mL)Result
P. aeruginosaaATCC 10145 [ MDB strain BU 277 type strain]1.0E+04Pseudomonas aeruginosa
Detected
ATCC BAA-1744 [109246]1.0E+04
ATCC 19429 [NCTC 6750]1.0E+04
ATCC 27853 [Boston 41501]1.0E+04
AR-Bank #00541.0E+04
AR-Bank #00921.0E+04
AR-Bank #01001.0E+04
AR-Bank #01031.0E+04
AR-Bank #01111.0E+04
Creighton University PS281.0E+04
NCTC 134371.0E+04

ª P. aeruginosa ATCC 25619 was not detected at any concentration tested. See Table 45Error! Reference source not found. for limitation.

Table 67. Serratia marcescens Isolates Tested and Detected

OrganismSource [Strain]Test concentration (copies/mL)Result
S. marcescensATCC 13880 [Type strain]1.0E+04Serratia marcescens
Detected
ATCC 27137 [CDC 3100-71]1.0E+04
ATCC 43297 [3G]1.0E+04
ATCC BAA-885 [Type strain KRED]1.0E+04
GRE 16590051.0E+04
GRE 16590041.0E+04
JMI 6971.0E+04

Table 68. Staphylococcus aureus Isolates Tested and Detected

| Organism | Source [Strain] [PFGE Type if applicable] | Test concentration
(copies/mL) | Result |
|------------------|--------------------------------------------------------------|-----------------------------------|-----------------------------------|
| | Staphylococcus aureus representing PFGE Types USA100-USA1200 | | |
| S. aureus | NARSA NRS705 [PFGE USA100] | 1.0E+04 | Staphylococcus aureus
Detected |
| | NARSA NRS701 [PFGE USA200] | 1.0E+04 | |
| | ATCC BAA-1717 [PFGE USA300] | 1.0E+05a | |
| | NARSA NRS683 [PFGE USA300] | 1.0E+04 | |
| | NARSA NRS662 [PFGE USA300] | 1.0E+04 | |
| | NARSA NRS707 [PFGE USA300] | 1.0E+04 | |
| | ATCC BAA-1707 [PFGE USA400] | 1.0E+04 | |
| | NARSA NRS691 [PFGE USA500] | 1.0E+04 | |
| | NARSA NRS648 [PFGE USA600] | 1.0E+04 | |
| | NARSA NRS689 [PFGE USA700] | 1.0E+04 | |
| | NARSA NRS668 [PFGE USA800] | 1.0E+04 | |
| | ATCC BAA-1749 [PFGE USA900 96:308] | 1.0E+04 | |
| | ATCC BAA-1759 [PFGE USA900 N7129] | 1.0E+04 | |
| | ATCC BAA-1700 [PFGE USA1000] | 1.0E+04 | |
| | BELNP 46081 [PFGE USA1100 HIP12899] | 1.0E+04 | |

49

| Organism | Source [Strain] [PFGE Type if applicable] | Test concentration
(copies/mL) | Result |
|------------------------------------------------------------|------------------------------------------------------------|-----------------------------------|--------|
| | ATCC BAA-1765 [PFGE USA1200 102-04] | 1.0E+04 | |
| | ATCC BAA-1691 [Not USA100-1100] | 1.0E+04 | |
| Methicillin Sensitive Staphylococcus aureus (MSSA) | ATCC 10832 [Wood 46] | 1.0E+04 | |
| | ATCC 14154 [Rose] | 1.0E+04 | |
| | ATCC 12600 [NCTC Type strain] | 1.0E+04 | |
| | ATCC 25923 [Seattle/1945] | 1.0E+04 | |
| | ATCC 29213 [Wichita] | 1.0E+04 | |
| | ATCC BAA-2421 [Mass/2010] | 1.0E+04 | |
| | Rennes 1060728 | 1.0E+04 | |
| | GRE 1062519 [SCCmec Type: III / MREJ xix]b | 1.0E+04 | |
| | Borderline Resistant Staphylococcus aureus (BORSA) | | |
| | SUN1 [Sunnybrook] | 1.0E+04 | |
| Methicillin Resistant Staphylococcus aureus (MRSA) | ATCC 43300 [F182 Kansas / SCCmec Type: II] | 1.0E+04 | |
| | ATCC BAA-2422 [Worcester MA/2010 / SCCmec Type: II] | 1.0E+04 | |
| | ATCC BAA-1720 [MRSA252 / SCCmec Type: II / PFGE
USA200] | 1.0E+04 | |
| | NARSA NRS745 [CA-629 / SCCmec Type: V] | 1.0E+04 | |
| | ATCC BAA-38 [E2125 / SCCmec Type: I] | 1.0E+04 | |
| | NARSA NRS686 [MREJ type i] | 1.0E+04 | |
| | ATCC BAA-44 [HPV107 / SCCmec Type: I / PFGE: Iberian] | 1.0E+04 | |
| | ATCC BAA-41 [NYBK2464 / SCCmec Type: II / PFGE 100] | 1.0E+04 | |
| | NARSA NRS385 [MREJ type ii] | 1.0E+04 | |
| | ATCC BAA-42 [HDE288 / SCCmec: Type VI / PFGE 800] | 1.0E+04 | |
| | ATCC BAA-39 [HUSA304 / SCCmec Type: III] | 1.0E+04 | |
| | ATCC BAA-40 [CPS22 / SCCmec Type: III] | 1.0E+04 | |
| | GRE 1062264 [SCCmec Type: IV / MREJ type iv] | 1.0E+04 | |
| | GRE 1055015 [SCCmec Type: IVa / MREJ type vi] | 1.0E+04 | |
| | GRE 0759084 [SCCmec Type: IV / MREJ type v] | 1.0E+04 | |
| | GRE 0860042 [SCCmec Type: III / MREJ type vii] | 1.0E+04 | |
| | GRE 1052034 [MREJ ix] | 1.0E+04 | |
| | GRE 1151100 [SCCmec Type: IV / MREJ type xi] | 1.0E+04 | |
| | GRE 0960006 [MREJ type xii] | 1.0E+04 | |
| | GRE 1055017 [SCCmec Type: IVa / MREJ type xiii] | 1.0E+04 | |
| | GRE 0759163 [MREJ type xiv] | 1.0E+04 | |
| | GRE 1062373 [MREJ type xv] | 1.0E+04 | |
| | GRE 1057114 [MREJ type xvii] | 1.0E+04 | |
| | GRE 1062292 [MREJ type xviii] | 1.0E+04 | |
| Methicillin Resistant Staphylococcus aureus (MRSA) - mecC+ | ATCC BAA-2312 [M10/0061 / SCCmec Type: XI / mecC] | 1.0E+04 | |
| | ATCC BAA-2313 [M10/0148 / SCCmec Type: XI / mecC] | 1.0E+04 | |

ª Staphylococus aureus ATCC BAA-1717 was not detected at 1.0E+04 copies/mL with accurate bir results. The expected detection and 104 bin result was reported in ≥50% of additional replicates tested at 1.0E+04 opies in the initiation on reactivity could be identified based on the isolate sequence.

b MREJ type xix characterized as MSSA129.

Table 69. Streptococcus aqalactiae Isolates Tested and Detected

| Organism | Source [Strain] | Test concentration
(copies/mL) | Result |
|---------------|--------------------------------------|-----------------------------------|--------------------------|
| S. agalactiae | NCTC 8017 [MK 104 P] | 1.0E+04 | |
| | ATCC 13813 [Ia/c Type Strain] | 1.0E+04 | |
| | ATCC 12403 [III Typing Strain D136C] | 1.0E+04 | Streptococcus agalactiae |
| | ATCC 12386 [Grouping strain O90R] | 1.0E+04 | Detected |
| | ATCC BAA-611 [V 2603 V/R] | 1.0E+04 | |
| | ATCC BAA-2669 [VIII 5030-08] | 1.0E+04 | |
| | Clinical Isolate [Utah/2010/CI03] | 1.0E+04 | |

Table 70. Streptococcus pneumoniae Isolates Tested and Detected
---------------------------------------------------------------------

| Organism | Serotype | Source [Strain] | Test concentration
(copies/mL) | Result |
|---------------|----------------|-----------------------------|-----------------------------------|--------------------------|
| S. pneumoniae | 3 | ATCC 6303 | 1.0E+04 | |
| | 5 | ATCC BAA-341 [SPN1439-106] | 1.0E+04 | |
| | 11A | NCTC 11900 [Gorman] | 1.0E+04 | Streptococcus pneumoniae |
| | 14 | ATCC 700672 [VH14] | 1.0E+04 | Detected |
| | 19A | ATCC 700673 [Hungary 19A-6] | 1.0E+04 | |
| | Non-capsulated | ATCC BAA-255 [R6] | 1.0E+04 | |

50

unknownATCC BAA-1409 [62076]1.0E+04
---------------------------------------------

| Organism | Source [Strain] | Test concentration
(copies/mL) | Result |
|-------------|---------------------------------------------------|-----------------------------------|------------------------------------|
| S. pyogenes | ATCC 12344 [Typing strain T1, NCIB 11841, SF 130] | 1.0E+04 | |
| | ATCC 12348 [Typing strain S43 Type 6] | 1.0E+04 | |
| | ATCC 12384 [Typing strain C203 Type 3] | 1.0E+04 | |
| | ATCC 19615ª [Bruno] | 1.0E+06ª | Streptococcus pyogenes
Detected |
| | ATCC 700294 [SF370; M1 GAS [M-type 1 T-type 1]] | 1.0E+05b | |
| | ATCC 49399 [QC A62] | 1.0E+04 | |
| | ATCC BAA-595 [MGAS 315, serotype M3] | 1.0E+04 | |
| | ATCC BAA-947 [MGAS 5005, serotype M1] | 1.0E+04 | |

Table 71. Streptococcus pyogenes Isolates Tested and Detected

a See Table 45 Error! Reference source not found. for limitation.

® Sireplococcus pyogenes ATCC 700294 was detected in 3/5 replicates at 1.0E+04 copies/mL bin results and 3/3 replicates at 1.0E+05 copies/mL with 10°5 copies/mL bin results. The expected in ≥50% of additional replicates tested at 1.0E+04 copies/mL and no limitation on reactivity could be identified based on isolate sequence.

The following tables (Table 72 - Table 79) describe the reactivity of the AMR genes assays with different AMR gene types in various host bacteria. Results are shown for the isolates tested as well as predictions of reactivity with untested AMR gene types based on in silico analysis of sequences retrieved from public databases from June 2016 to Sept 2016.

| Organism | Source [Strain] | Test concentration
(copies/mL) | Result |
|------------------|------------------------------------------------------------------------------------------------------------------------------|-----------------------------------|------------------------------------|
| S. aureus | Methicillin Sensitive Staphylococcus aureus (MSSA)
containing SCCmec cassette (non-functional mecA variant) | | |
| | ATCC BAA-2421 [Mass/2010] | 1.0E+04 | |
| | Methicillin Resistant Staphylococcus aureus (MRSA)
(Characterized SCCmec Types) | | |
| | NARSA NRS705 [NY-12 / SCCmec Type: II] | 1.0E+04 | |
| | NARSA NRS701 [MN-082 / SCCmec Type: II] | 1.0E+04 | |
| | ATCC BAA-1717 [TCH1516 / SCCmec Type: IVa] | 1.0E+05a | |
| | NARSA NRS683 [GA-298 / SCCmec Type: IV] | 1.0E+04 | |
| | NARSA NRS662 [CO-34 / SCCmec Type: IV] | 1.0E+04 | |
| | NARSA NRS707 [NY-155 / SCCmec Type: IV] | 1.0E+04 | |
| | ATCC BAA-1707 [MW2 / SCCmec Type: IV] | 1.0E+04 | |
| | NARSA NRS691 [GA-62 / SCCmec Type: IV] | 1.0E+04 | |
| | NARSA NRS648 [CA-347 / SCCmec Type: II or IV] | 1.0E+05a | |
| | NARSA NRS689 [GA-442 / SCCmec Type: IV] | 1.0E+04 | |
| | NARSA NRS668 [CO-72 / SCCmec Type: IV] | 1.0E+04 | mecA/C and MREJ
Detected |
| | ATCC BAA-1700 [HFH-33798 / SCCmec Type: IVb] | 1.0E+04 | |
| | BEI NR-46081b (NRSA NRS484) [HIP12899 / SCCmec Type: IV] | 1.0E+05a | |
| | ATCC BAA-1691 [HFH-30137 / SCCmec Type: IV] | 1.0E+04 | |
| | ATCC 43300 [F182 Kansas / SCCmec Type: II ] | 1.0E+04 | |
| | ATCC BAA-2422 [Worcester MA/2010 / SCCmec Type: II] | 1.0E+04 | |
| | ATCC BAA-1720 [MRSA252 / SCCmec Type: II] | 1.0E+04 | |
| | NARSA NRS745 [CA-629 / SCCmec Type: IV or V] | 1.0E+04 | |
| | Methicillin Resistant Staphylococcus aureus (MRSA)
(Characterized MREJ Types) | | |
| | ATCC BAA-38 [MREJ type i] | 1.0E+04 | |
| | NARSA NRS686 [MREJ type i] | 1.0E+04 | |
| | ATCC BAA-44 [MREJ type ii] | 1.0E+04 | |
| | ATCC BAA-41 [MREJ type ii] | 1.0E+04 | |
| | NARSA NRS385 [MREJ type ii] | 1.0E+04 | |
| | ATCC BAA-42 [MREJ type ii] | 1.0E+04 | |
| | ATCC BAA-39 [MREJ type iii] | 1.0E+04 | |
| | ATCC BAA-40 [MREJ type iv] | 1.0E+04 | |
| | GRE 1062264 [MREJ type iv] | 1.0E+04 | |
| | GRE 1055015 [MREJ type vi] | 1.0E+04 | |
| | GRE 0860042 [MREJ type vii] | 1.0E+04 | |
| | GRE 1052034 [MREJ type ix] | 1.0E+04 | |
| | GRE 1151100 [MREJ type xi] | 1.0E+04 | |
| | GRE 0960006 [MREJ type xii] | 1.0E+04 | |
| | GRE 1055017 [MREJ type xiii] | 1.0E+04 | |

Table 72. Isolates Containing mecA/C and MREJ Tested and Detected

51

| Organism | Source [Strain] | Test concentration
(copies/mL) | Result | |
|---------------------|-----------------------------------------------------------------------|-----------------------------------|----------------------------------------|--|
| | GRE 0759163 [MREJ type xiv] | 1.0E+04 | | |
| | GRE 1062373 [MREJ type xv]c | 1.0E+06b | | |
| | GRE 1057114 [MREJ type xvii] | 1.0E+04 | | |
| | GRE 1062292 [MREJ type xviii]c | 3.3E+08 | | |
| | GRE 1062519 [MREJ type xix]c,d | 1.0E+07 | mecA/C and MREJ Not
Detected | |
| | Methicillin Resistant Staphylococcus aureus (MRSA) | | | |
| | ( SCCmec Type: XI / mecC / mecA LGA251 variants) | | | |
| | ATCC BAA-2312 [M10/0061 / SCCmec Type: XI / mecC ] | 1.0E+04 | mecA/C and MREJ | |
| | ATCC BAA-2313 [M10/0148 / SCCmec : Type XI / mecC ] | 1.0E+04 | Detected | |
| S. argenteus | Methicillin Resistant Staphylococcus argenteus | | | |
| | DSM 28299 [MSHR-1132] | 1.00E+05 | | |

2 mecA/C and MREJ assays positive in less than three replicates at 1.0E+04 copies/mL, no sequence based limitation on reactivity identified.

b Bacteria obtained through NARSA for distribution by BEI Resources, NIAID, NIH: Staphylococcus aureus, Strain HIP12899, NR-46081

& See Table 45Error! Reference source not found. for limitation.

d MREJ type xix characterized MSSA129.

Table 73. In Silico Reactivity Predictions for mecA/C and MREJ

mecA/C a,bMREJd
DetectedReduced Reactivity
or Not DetectedDetectedReduced Reactivity
or Not DetectedUnknown
Reactivity
(no sequences)
mecA in S. aureus cmecA in some isolates
of S. capitis, S. kloosii
and S. vitulinusMREJ i, ia – viieMREJ ixfMREJ viii
mecC in S. aureusmecC in S. sciuriMREJ xi-xivMREJ xvgMREJ x
mecA and mecC in
non-aureus staphylococci
(including S. argenteus )MREJ xvi – xviiiMREJ xviii
MREJ in
S. argenteusMREJ xix, xxh
MREJ in non-aureus
staphylococci and
other speciesd

" July 2016; analysis of 1,257 database mecA sequences from S. aureus, as well as well as meet and mec sequences from non-aureus staphylococci.

b mecC is also referenced as SCCmec XI and mecALGA251.

& Limited or reduced reactivity predicted for 2/1,257 mecA sequences from S. aureus (0.2%).

4 June 2016; analysis of approximately 1,450 typed MREJ database sequences from S. aureus, non-aureus staphylococi and non-staphylococus species (Bacillus thuringiensis, Macrococcus casedyticus, Clostridium aciduric, and Rummeliibacillus stabekisii).

· Limited or reduced reactivity predicted for 1/14); normal reactivity observed for the isolate of MREJ iii tested (see Table 72Error! Reference source not found.).

f Limited or reduced reactivity predicted for 2/8 MREJ ix sequences (25.0%); normal reactivity observed for the isolate 72)

6 Reduced reactivity predicted by in silico analysis and observed with the isolate of MREJ xv tested (see Table 72Error! Reference source not found.).

4 MREJ xix and xx were not included in the assay design because they were identified from methicillin-sensitive S. aureus'22

Table 74. Isolates Containing the black.agene Tested and In Silico Reactivity Predictions

| CTX-M Type | Organism | Source | Test concentration
(copies/mL) | Result |
|-----------------------------------|---------------|---------------------|-----------------------------------|--------------------------------------|
| Isolates Tested | | | | |
| | E. coli | AR-Bank #0137ª | 1.0E+04 | CTX-M Detected |
| | K. oxytoca | GRE 1254054 | 1.0E+04 | |
| CTX-M | K. pneumoniae | AR-Bank #0068ª | 1.0E+04 | |
| | K. pneumoniae | AR-Bank #0153ª | 1.0E+04 | |
| | | GRE 1355030 | 1.0E+04 | |
| CTX-M-1 | E. coli | AR-Bank #0162 | 1.0E+04 | CTX-M Detected |
| CTX-M-2 | K. pneumoniae | AR-Bank #0107 | 1.0E+04 | |
| CTX-M-8 | K. aerogenes | GRE 1254066 | 1.0E+04 | |
| CTX-M-9 | E.coli | AR-Bank #0086 | 1.0E+04 | |
| | E.cloacae | NCTC 13464 | 1.0E+04 | |
| CTX-M-14 | K. pneumoniae | AR-Bank #0079 | 1.0E+04 | |
| CTX-M-15 | E.coli | Zeptometrix 0801905 | 1.0E+04 | |
| CTX-M-22 | P. mirabilis | GRE 1254053 | 1.0E+04 | |
| CTX-M-25 | K. pneumoniae | NCTC 13465 | 1.0E+04 | |
| In silico Reactivity Predictionsª | | | | |
| Detected | | Not Detected | | Unknown Reactivity
(no sequences) |

52

CTX-M-1 – CTX-M-117CTX-M-150CTX-M-151CTX-M-118CTX-M-143
CTX-M-121 – CTX-M-126CTX-M-152CTX-M-119CTX-M-145
CTX-M-129 – CTX-M-132CTX-M-155 – CTX-M-177CTX-M-120CTX-M-146
CTX-M-134CTX-M-179 – CTX-M-185CTX-M-127CTX-M-149
CTX-M-136 – CTX-M-139CTX-M-128CTX-M-153
CTX-M-141 – CTX-M-142CTX-M-133CTX-M-154
CTX-M-144CTX-M-135CTX-M-178
CTX-M-147 – CTX-M-148CTX-M-140

a July 2016; analysis of over 1,200 database CTX-M sequences (typed and untyped).

Table 75. Isolates Containing the blamp gene Tested and In Silico Reactivity Predictions

IMP TypeOrganismSourceTest concentration (copies/mL)Result
Isolates Tested
IMPK. aerogenesAR-Bank #0161$1.0E+04$
E. coliGRE 1062016$1.0E+04$
K. pneumoniaeAR-Bank #0080$1.0E+04$
IMP-1aP. aeruginosaAR-Bank #0103$1.0E+04$
IMP-3aE. coliGRE 1252008$1.0E+04$IMP Detected
IMP-4A. baumanniiGRE 1062081$1.0E+04$
IMP-8K. pneumoniaeGRE 1062084$1.0E+04$
IMP-9E. coliGRE 1252009$1.0E+04$
IMP-14P. aeruginosaAR-Bank #0092$1.0E+04$
In silico Reactivity Predictionsb
DetectedReduced Reactivity
or Not DetectedUnknown Reactivity
(no sequences)
IMP-1 – IMP-30aIMP-31IMP-36
IMP-40 – IMP-45IMP-35IMP-39
IMP-32 – IMP-34IMP-46
IMP-48 – IMP-49IMP-47
IMP 37 – IMP-38IMP-50
IMP-51 – IMP-56IMP-57
IMP-58 – IMP-60

ª Limited or reduced reactivity predicted for 1/36 (2.8%) of IMP-1 and 1/3 (33.3%) of IMP-3 sequences.

b June 2016; analysis of over 220 database IMP sequences (typed and untyped).

Table 76. Isolates Containing the blake gene Tested and Detected, and In Silico Reactivity Predictions

| KPC Type | Organism | Source | Test concentration
(copies/mL) | Result |
|-----------------------------------|---------------|---------------------------|--------------------------------------|------------------|
| | | Isolates Tested | | |
| KPC | E. cloacae | ATCC BAA-2341 | 1.0E+04 | KPC Detected |
| | E. hormaechi | ATCC BAA-2082 | 1.0E+04 | |
| | P. mirabilis | AR-Bank #0156 | 1.0E+04 | |
| | K. oxytoca | AR-Bank #0147 | 1.0E+04 | |
| | K. pneumoniae | AR-Bank #0097 | 1.0E+04 | |
| | K. oxytoca | JMI 2523 | 1.0E+04 | |
| | K. oxytoca | JMI 7818 | 1.0E+04 | |
| KPC-2 | K. pneumoniae | Zeptometrix 0801886 | 1.0E+04 | |
| | K. pneumoniae | JMI 328 | 1.0E+04 | |
| | K. pneumoniae | ATCC BAA-1705 | 1.0E+04 | |
| | S. marcescens | JMI 697 | 1.0E+04 | |
| | E. coli | AR-Bank #0061 | 1.0E+04 | |
| KPC-3 | K. oxytoca | JMI 2661 | 1.0E+04 | |
| KPC-4 | K. pneumoniae | JMI 766 | 1.0E+04 | |
| KPC-5 | P. aeruginosa | Creighton University PS28 | 1.0E+04 | |
| In silico Reactivity Predictionsa | | | | |
| Detected | | Not Detected | Unknown Reactivity
(no sequences) | |
| KPC-1-19 | KPC-21-22 | KPC-24-26 | None | KPC-20
KPC-23 |

a August 2016; analysis of approximately 1,125 database KPC sequences (typed and untyped).

Table 77. Isolates Containing the blawn gene Tested and In Silico Reactivity Predictions

| NDM Type | Organism | Source | Test concentration
(copies/mL) | Result |
|-----------------|---------------|---------------|-----------------------------------|--------------|
| Isolates Tested | | | | |
| NDM | E. coli | AR-Bank #0162 | 1.0E+04 | |
| NDM | K. pneumoniae | AR-Bank #0153 | 1.0E+04 | NDM Detected |
| NDM | K. pneumoniae | AR-Bank #0068 | 1.0E+04 | |

53

| NDM Type | Organism | Source | Test concentration
(copies/mL) | Result |
|-----------------------------------|--------------|---------------|-----------------------------------|--------|
| NDM-1ª | P. mirabilis | AR-Bank #0159 | 1.0E+04 | |
| NDM-2 | A. baumannii | AR-Bank #0033 | 1.0E+04 | |
| NDM-5 | A. baumannii | GRE 1153064 | 1.0E+04 | |
| NDM-6 | E. coli | AR-Bank #0150 | 1.0E+04 | |
| | E. coli | AR-Bank #0137 | 1.0E+04 | |
| In silico Reactivity Predictionsb | | | | |
| | Detected | | Not Detected | |
| NDM-1ª | NDM-7 | NDM-13 | None | |
| NDM-2 | NDM-8 | NDM-14 | | |
| NDM-3 | NDM-9 | NDM-15 | | |
| NDM-4 | NDM-10 | NDM-16 | | |
| NDM-5 | NDM-11 | | | |
| NDM-6 | NDM-12 | | | |

ª Limited or reduced reactivity is predicted for 3/430 NDM-1 sequences (0.7%).

b June 2016; analysis of 900 database NDM sequences (typed and untyped).

Table 78. Isolates Containing the blacks and like genes Tested and In Silico Reactivity Predictions

| OXA-48-like Typea | Organism | Source | Test concentration
(copies/mL) | Result |
|-----------------------------------|----------------------|---------------|-----------------------------------|----------------------|
| Isolates Tested and Detected | | | | |
| OXA-48 | K. aerogenes | AR-Bank #0074 | 1.0E+04 | |
| OXA-48-like | S. marcescens | GRE 1411136 | 1.0E+04 | OXA-48-like Detected |
| OXA-162 | S. marcescens | GRE 1411137 | 1.0E+04 | |
| OXA-181 | K. pneumoniae | GRE 1355030 | 1.0E+04 | |
| OXA-232 | K. pneumoniae | AR-Bank #0068 | 1.0E+04 | |
| | K. pneumoniae | AR-Bank #0075 | 1.0E+04 | |
| In silico Reactivity Predictionsa | | | | |
| Detected | | | Not Detectedb | |
| OXA-48 | OXA-204 | OXA-370 | OXA-163° | OXA-438° |
| OXA-48-like | OXA-232 | OXA-484 | OXA-247° | OXA-439° |
| OXA-162 | OXA-244 | OXA-505 | OXA-405° | |
| OXA-181 | OXA-245 | | OXA-416 | |
| OXA-199 | OXA-252 | | OXA-436° | |

a June 2016; analysis of 165 database OXA-48-like sequences (typed and untyped).

  • Sequence analysis predicts that the listed OXA-48-like types will not be detected. Non-OXA-48-like, OXA-31-ike, OXA-51like, and OXA-58-like, OXA-143a-like and OXA-143-like) will also not be detected.

& Deletion variants with altered carbapenem hydrolysis activity, as described for OXA-163130.

Table 79. Isolates Containing the blaw gene Tested and In Silico Reactivity Predictions

| VIM Type | Organism | Source | Test concentration
(copies/mL) | Result |
|-----------------------------------|-----------------|-----------------|---------------------------------------|--------------------------------------|
| Isolates Tested | | | | |
| VIM | E. cloacae | AR-Bank #0154 | 1.0E+04 | |
| VIM | P. aeruginosa | AR-Bank #0111 | 1.0E+04 | |
| VIM | K. pneumoniae | AR-Bank 0076 | 1.0E+04 | |
| VIM-2a | P. aeruginosa | AR-Bank #0100 | 1.0E+04 | VIM Detected |
| VIM-4 | P. aeruginosa | AR-Bank #0054 | 1.0E+04 | |
| VIM-7 | E. coli | GRE 1256018 | 1.0E+04 | |
| VIM-10 | P. aeruginosa | NCTC 13437 | 1.0E+04 | |
| In silico Reactivity Predictionsb | | | | |
| Detected | | | Reduced Reactivity
or Not Detected | Unknown Reactivity
(no sequences) |
| VIM-1 – VIM-20a | VIM-23 – VIM-47 | VIM-49 – VIM-51 | VIM-39
VIM-45
VIM-46 | VIM-21
VIM-22
VIM-48 |

a Limited or reduced reactivity is predicted for 2/177 VIM-2 sequences (1.1%).

b September 2016; analysis of over 600 database VIM sequences (typed and untyped).

Analytical Specificity (Cross-Reactivity and Exclusivity)

There is a risk of false positive results due to non-specific amplification and/or cross-reactivity with organisms found in the respiratory tract. The potential for non-specific amplification and detection by BioFire Diagnostics 510(k) FilmArray Pneumonia Panel

54

the FilmArray Pneumonia Panel assays was evaluated by in silico analyses of available sequences and by empirical (wet) testing of high concentrations of organisms in contrived samples. The observed and predicted cross-reactivities for organisms closely related to those detected by the panel and unrelated organisms that may be present in lower respiratory specimens are summarized in Table 80. Erroneous results due to cross-reactivity with organisms that were not evaluated or new variant sequences that emerge is also possible.

On-panel organisms were tested to assess the potential for intra-panel cross-reactivity (Table 81Error! Reference source not found.). Off-panel organisms included species of the same genus or otherwise genetically related to organisms detected by the panel, as well as normal flora and pathogens that may be present in sputum-like and BAL-like specimens (Table 82). Antimicrobial resistance genes were also evaluated in conjunction with on and off panel host organisms.

The final concentration of analyte in the sample (typically >1.0E+07 CFU/mL for bacteria and fungi and >1.0E+05 TCID30/mL for viruses) represented levels ~100 - 100,000 fold higher than the LoD or lowest reportable level of the FilmArray Pneumonia Panel assays.

FilmArray Pneumonia Panel ResultCross-Reactive Organism
Closely-Related Species
Escherichia coliEscherichia fergusoniia
Klebsiella oxytocaShigella species (S. boydii, S. dysenteriae, S. flexneri, S. sonnei)a
Staphylococcus aureusKlebsiella michiganensisa
Pseudomonas aeruginosaStaphylococcus argenteusb
Staphylococcus schweitzeric
Pseudomonas putidad
Unrelated Species
Human Rhinovirus/EnterovirusBordetella speciese
Parainfluenza VirusfAspergillus niger
Cryptococcus laurentii
AdenovirusCryptococcus uniguttulatus
CTX-MhStenotrophomonas acidaminiphilag
Acinetobacter schindleri
Burkholderia vietnamiensisi
Escherichia colij,kLelliottia amnigena (Enterobacter amnigenus)
Enterobacter kobei
Enterobacter ludwigii
Enterobacter cloacae

a Genetically or phenotypically indistinguished by standard laboratory techniques. Detected at a concentrations ≥1.0E+04 copies/mL.

b Genetically or phenotypically indistinguished by standard laboratory techniques. Detected at a concentrations ≥1.0E+05 copies/mL

& Genetically or phenotypically indistinguished by standard laboratory techniques. Detected at a concentrations ≥1.0E+06 copies/mL.

d Cross-reactivity possible at concentrations >1.0E+07 copies/mL.

6 Cross-reactivity with B. pertussis confirmed at ≥1.0E+06 CFUmL. Cross-reactivity with B. parapertusis was not observed at 1.0E+08 CFU/mL, but possible based on sequence analysis.

F Cross-reactivity was observed with A. niger, C. lawentulatus at concentrations >1.0E+06 copiesmL. Cross-reactivity with other Cryptococcus species may be possible based on sequence analysis.

$ S. acidaminiphila has not been isolated from human clinical specimens, no cross-reactivity observed with other Steners species.

h Cross-reactive product observed only at concentrations >4.5E+07 CFUmL and only reported if an applicable gram-negative bacteried. 1 Not tested. Predicted by in silico analysis.

i If observed, results will be reported as Escherichia coli 10^4 copies/mL

" Based on in silico analysis, cross-reactivity is also possible at high copies/mL) of other Enterobracter species (E. hornachei, K. aerogenes, E. lingolyticus), Citrobacter koseri, Escherichia vulneris, and Leclercia adecaboxylata.

Table 81. On Panel Organisms Tested for Evaluation of FilmArray Pneumonia Panel Analytical Specificity False positive results were observed when testing the species shown in bold.

55

ON-PANEL
Bacteria
Acinetobacter baumanniiEnterobacter kobei aKlebsiella quasipneumoniaePseudomonas aeruginosa
Acinetobacter calcoaceticusEnterobacter ludwigii aKlebsiella variicolaSerratia marcescens
Acinetobacter nosocomialisEscherichia coliMoraxella catarrhalisStaphylococcus aureus
Acinetobacter pittiiHaemophilus influenzaeProteus hauseriStreptococcus agalactiae
Enterobacter asburiaeKlebsiella aerogenesProteus mirabilisStreptococcus pneumoniae
Enterobacter cloacae aKlebsiella oxytocaProteus penneriStreptococcus pyogenes
Enterobacter hormaecheiKlebsiella pneumoniaeProteus vulgaris
Atypical Bacteria
Chlamydia pneumoniaeLegionella pneumophilaMycoplasma pneumoniae
Viruses
Adenovirus BCoronavirus NL63Human MetapneumovirusParainfluenza Virus 2
Adenovirus CCoronavirus OC43Influenza AParainfluenza Virus 3
Adenovirus EEnterovirusInfluenza BParainfluenza Virus 4
Coronavirus 229EHuman RhinovirusParainfluenza Virus 1Respiratory Syncytial Virus
Coronavirus HKU1
Antimicrobial Resistance Genes
CTX-M
( Klebsiella oxytoca )KPC
( Klebsiella pneumoniae )OXA-48-like
( Serratia marcescens )mecA and MREJ
( Staphylococcus aureus )
IMP
( Pseudomonas aeruginosa )NDM
( Acinetobacter baumannii )VIM
( Enterobacter cloacae )

ª See Table 80Error! Reference source not found. for cross-reactivity information.

Table 82. Off-Panel Bacteria Tested or Evaluated by In Silico Analysis for FilmArray Pneumonia Panel Analytical Specificity

False positive results were observed when testing the species shown in bold.

OFF-PANEL
Bacteria
Abiotrophia defectivaEscherichia fergusoniiaMycobacterium tuberculosisShigella boydii a
Achromobacter
xylosoxidansEscherichia hermaniiMycoplasma bovisShigella dysenteriaea
Acinetobacter haemolyticusEscherichia vulnerisMycoplasma genitaliumShigella flexneria
Acinetobacter johnsoniiFluoribacter dumoffeiMycoplasma hominisShigella sonneia
Acinetobacter juniiFusobacterium variumMycoplasma oraleStaphylococcus argenteusa
Acinetobacter lwolfiiGemella morbillorumNeisseria gonorrhoeaeStaphylococcus capitis
Acinetobacter radioresistensGranulicatella adiacensNeisseria lactamicaStaphylococcus caprae
Acinetobacter schindleriaaHaemophilus ducreyiNeisseria meningitidisStaphylococcus cohnii
Acinetobacter ursingiiHaemophilus haemolyticusNeisseria mucosaStaphylococcus haemolyticus
Actinobacillus
actinomycetemcomitansHaemophilus parahaemolyticusNeisseria siccaStaphylococcus epidermidis
(mecA)
Actinobacillus hominisHaemophilus parainfluenzaeNocardia asteroidesStaphylococcus hominis
Actinobacillus ureaeHaemophilus parasuisNocardia brasilensisStaphylococcus intermedius
Actinomyces isrealiiHaemophilus sputorumbPantoea agglomeransStaphylococcus lugdunensis
Actinomyces naeslundiiHafnia alveiPasteurella multocidaStaphylococcus lutrae
Bacillus cereusHafnia paralveiPediococcus acidilacticiStaphylococcus pasteuri
Bacteriodes fragilisHelicobacter pyloriPeptostreptococcus anaerobiusStaphylococcus pseudointermedius
Bordatella bronchisepticaKingella kingaePluralibacter gergoviaeStaphylococcus saprophyticus
Bordatella parapertussisKlebsiella michiganensisaPorphyromonas gingivalisStaphylococcus schleiferi
Bordatella pertussisaKluyvera intermediaPrevotella intermediaStaphylococcus schweitzeria
Burkholderia cepaciaKluyvera ascorbataPrevotella melaninogenicaStaphylococcus sciuri
Burkholderia malleiLactobacillus acidophilusPrevotella oralisStaphylococcus warneri
Burkholderia multivoransLeclercia adecarboxylataPropionibacterium acnesStaphylococcus xylosus
Burkholderia pseudomalleiLegionellaProvidencia rettgeriStenotrophomonas
bozemanii(OXA-48-like)acidaminiphilaa
Cardiobacterium hominisLegionella cincinnatiensisProvidencia stuartiiStenotrophomonas maltophilia
Cedecea davisaeLegionella feeleiiPseudomonas fluorescensStenotrophomonas nitritireducens
Chlamydia trachomatisLegionella lansingensisPseudomonas luteolaStenotrophomonas rhizophila
Chlamydophila psittaciLegionella longbeachaePseudomonas nitroreducensStreptococcus equi subsp.
Zooepidemicus
Citrobacter freundii (KPC)Legionella micdadeiPseudomonas oryzihabitansStreptococcus mitis
Citrobacter koseri
(OXA-48-like)Legionella wadsworthiiPseudomonas pertucinogenaStreptococcus mutans
Citrobacter sedlakiiLelliottia nimipressuralisPseudomonas putidaa (IMP)Streptococcus oralis
Citrobacter werkmanii (VIM)Lelliottia amnigenaa
(Enterobacter amnigenus)Pseudomonas stutzeriStreptococcus parasanguinis
Clostridium difficileLeuconostoc lactisRalstonia pickettiiStreptococcus pseudopneumoniae
Clostridium perfringensListeria monocytogenesRaoultella ornithinolyticaStreptococcus salivarius
Corynebacterium diphtheriaeMacrococcus caseolyticusRaoultella planticolaStreptococcus sanguinis

56

OFF-PANEL
Corynebacterium genitaliumMicrococcus luteusRaoultella terrigenaStreptococcus tigurinus
Corynebacterium
pseudodiptherticumMoraxella equiRhodococcus equiStreptomyces anulatusc
Corynebacterium urealyticumMoraxella lacunataRothia mucilaginosaTreponema denticola
Cronobacter sakazakiiMoraxella lincolniiSalmonella enterica (CTX-M)Ureaplasma parvum
Eikenella corrodensMoraxella nonliquiefaciensSerratia fonticolaUreaplasma urealyticum
Enterobacter cancerogenusMorganella morganii (NDM)Serratia liquefaciensVagococcus fluvialis
Enterobacter massiliensisMycobacterium africanumbSerratia odoriferaVeillonella parvula
Enterobacter soliMycobacterium bovisSerratia plymuthicaYersinia enterocolitica
Enterococcus faeciumMycobacterium capraecSerratia rubidaeaYersinia pseudotuberculosis
Enterococcus faecalisMycobacterium microtib
Viruses
BocavirusHantavirusHuman Papillomavirus (HPV)Mumps virus
CytomegalovirusHerpes simplex virus 1Influenza CVaricella zoster virus
Epstein Barr virusHuman Immunodeficiency Virus (HIV)Middle East Respiratory Syndrome Coronavirus (MERS-CoV)Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV)
German Measles Virus (Rubella)Measles Virus (Rubeola)
Fungi/Yeast
Aspergillus flavusCoccidioides posadasiiFusarium kyushensePneumocystis carinii
Aspergillus fumigatusCryptococcus albidusHistoplasma capsulatumPneumocystis jirovecii
Aspergillus nigeraCryptococcus gattiiPaecilomyces variottiiPneumocystis murina
Aspergillus terreusCryptococcus laurentiiaParacoccidodes brasiliensisRhizopus microsporus
Blastomyces dermatitidisCryptococcus neoformansPenicillium chrysogenumScedosporium apiospermum
Candida albicansCryptococcus uniguttalatusaPenicillium marneffeiScedosporium prolificans
Candida glabrataFilobasidium capsuligenum
Antimicrobial Resistance Genes
AmpC
(Klebsiella (Enterobacter)
aerogenes)OXA-24/40 (non-48-like)
(Acinetobacter baumannii)SME
(Serratia marcescens)TEM
(Escherichia coli)
CMY (II)
(Escherichia coli)SHV
(Klebsiella pneumoniae)SPM
(Pseudomonas aeruginosa)VAN
(Staphylococcus aureus)
ompK36 [SHV-12,OMPC]
(Klebsiella pneumoniae)SCCmec variant lacking mecA or mecC ( Staphylococcus aureus )d

a See Table 80Error! Reference source not found. for cross-reactivity information.

b Analytical specificity was evaluated only by in silco analysis of whole genome sequences in public database. No cross-reactive is predicted based on the sequences analyzed.

c Tested at a concentration less than 1.0E+07 CFUmL and also evaluated by in silico analysis. Cross-reactivity was not observed in testing nor prediced based on the sequences analyzed.

4 Methicillin-sensitive isolate of S. aureus (Rentes MREJ sequence but no mecA or mecC gene (empy cassette). Staphylococus aureus was reported as Detected and the mecA/C and MREJ result was Not Detected

Precision (Reproducibility)

Precision (Reproducibility) testing was performed with contrived BAL samples over multiple days at three laboratory locations (sites) on a combination of FilmArray 2.0 and FilmArray Torch systems. The testing incorporated a range of potential variation introduced by operator, system, instrument or Torch module, concentration and reagent lot, for a total of 30 tests per system and 90 total replicates per sample/concentration.

Evaluation of the reproducibility of Detected results for atypical bacteria and viruses included samples containing combinations of five different analytes, at Negative, Low Positive

57

(1×LoD), and Moderate Positive (3×LoD) concentrations. Negative results were obtained from samples that were not spiked with the analyte (see evaluation of precision for bacterial analytes below).

A summary of results (percent (%) agreement with the expected Detected or Not Detected result) for atypical bacteria and viruses (by site and system) is provided in Table 83Error! Reference source not found..

AnalyteConcentration TestedExpected ResultFilmArray Site AFilmArray 2.0 Site BFilmArray Torch Site CAll Sites/Systems [95% CI]
Atypical Bacteria
Chlamydia pneumoniaeNone
(No Analyte)Not Detected780/780
100%780/780
100%780/780
100%2,340/2,340
100%
[99.8%-100%]
Legionella pneumophila
Philadelphia-1
ATCC 33152Moderate Positive
3x LoD
$1.5E+03$ CFU/mLDetected30/30
100%30/30
100%30/30
100%90/90
100%
[96.0%-100%]
Low Positive
1x LoD
$5.0E+02$ CFU/mLDetected30/30
100%30/30
100%30/30
100%90/90
100%
[96.0%-100%]
None
(No Analyte)Not Detected720/720
100%720/720
100%720/720
100%2,160/2,160
100%
[99.8%-100%]
Mycoplasma pneumoniaeNone
(No Analyte)Not Detected780/780
100%780/780
100%780/780
100%2,340/2,340
100%
[99.8%-100%]
Viruses
Adenovirus
Species B Serotype 3
ZeptoMetrix 0810062CFModerate Positive
3x LoD
$3.0E+00$ TCID50/mLDetected30/30
100%30/30
100%30/30
100%90/90
100%
[96.0%-100%]
Low Positive
1x LoD
$1.0E+00$ TCID50/mLDetected30/30
100%30/30
100%30/30
100%90/90
100%
[96.0%-100%]
None
(No Analyte)Not Detected720/720
100%720/720
100%720/720
100%2,160/2,160
100%
[99.8%-100%]
CoronavirusNone
(No Analyte)Not Detected780/780
100%776/780
99.5%780/780
100%2,336/2,340
99.8%
[99.6%-100%]
Human Metapneumovirus
16 Type A1
ZeptoMetrix 0810161CFModerate Positive
3x LoD
$1.5E+02$ TCID50/mLDetected30/30
100%30/30
100%30/30
100%90/90
100%
[96.0%-100%]
Low Positive
1x LoD
$5.0E+01$ TCID50/mLDetected30/30
100%29/30
96.7%30/30
100%89/90
98.9%
[94.0%-100%]
None
(No Analyte)Not Detected720/720
100%720/720
100%720/720
100%2,160/2,160
100%
[99.8%-100%]
Human Rhinovirus/EnterovirusNone
(No Analyte)Not Detected779/780
99.9%780/780
100%779/780
99.9%2,338/2,340
99.9%
[99.7%-100%]
Influenza A
H3N2
A/Port Chalmers/1/73
ATCC VR-810Moderate Positive
3x LoD
$1.5E+00$ TCID50/mLDetected30/30
100%30/30
100%30/30
100%90/90
100%
[96.0%-100%]
Low Positive
1x LoD
$0.5E-01$ TCID50/mLDetected30/30
100%29/30
96.7%30/30
100%89/90
98.9%
[94.0%-100%]
None
(No Analyte)Not Detected720/720
100%720/720
100%720/720
100%2,160/2,160
100%
[99.8%-100%]
Influenza BNone
(No Analyte)Not Detected780/780
100%780/780
100%780/780
100%2,340/2,340
100%
[99.8%-100%]
Parainfluenza Virus
Type 2
ZeptoMetrix 0810015CFModerate Positive
3x LoD
$7.5E+01$ TCID50/mLDetected30/30
100%30/30
100%30/30
100%90/90
100%
[96.0%-100%]
AnalyteConcentration TestedExpected ResultAgreement with Expected ResultAll Sites/Systems [95% CI]
FilmArray
Site AFilmArray 2.0
Site BFilmArray Torch
Site C
Low Positive
1× LoD
$2.5E+01 TCID_{50}/mL$Detected30/30
100%30/30
100%30/30
100%90/90
100%
[96.0%-100%]
None
(No Analyte)Not Detected720/720
100%720/720
100%720/720
100%2,160/2,160
100%
[99.8%-100%]
Respiratory Syncytial VirusNone
(No Analyte)Not Detected780/780
100%780/780
100%780/780
100%2,340/2,340
100%
[99.8%-100%]

Table 83. Reproducibility of FilmArray Pneumonia Panel Atypical Bacteria and Virus Results

58

Precision for bacterial analytes was measured at each concentration as 1) precision of bin results and 2) reproducibility of analyte detection. When a sample containing one or more bacteria is tested repeatedly, the precision of the bin results (probability that each replicate will receive the same bin result) will vary based on the concentration of nucleic acid measured and the relation of that concentration to the limits of each bin. Bin precision may be as low as 50% for values at a bin limit and precision will increase (up to 90% or higher) as the distance of the measured value from a bin limit increases. The precision of FilmArray Pneumonia Panel bin results will follow the model illustrated in Figure 1Error! Reference source not found.:

  • 90% at a bin center (Scenario 1)

  • ~60 90% between a bin limit and bin center (Scenario 2) ●
  • ~50% at bin limits (Scenario 3)

Image /page/58/Figure/5 description: The image shows a graph of the probability of a reported bin result versus the measured value. The graph shows the probability of reporting a bin result for different measured values, with the bins labeled as 'Not Detected', '10^4', '10^5', '10^6', and '≥10^7'. Below the graph, there are three scenarios that show the mean measured value for different bins. Scenario 1 shows a mean measured value of 10^5.0 with a probability of ≥90%, scenario 2 shows a mean measured value of 10^5.25 with probabilities of ~75% and ~25%, and scenario 3 shows a mean measured value of 10^5.5 with probabilities of ~50% and ~50%.

Figure 1. Model for Precision of FilmArray Pneumonia Panel Bin Results

Top: The probability (0.0 – 1.0) of the same bin results for each replicate tested varies based on proximity of the measured value to a bin limit.

Bottom: Expected distribution of bin results at different mean measured values.

Samples containing bacteria and corresponding antimicrobial resistance (AMR) genes were tested at six different concentrations over the reportable range and below. A summary of the bin precision (percent (%) of replicates reported in each bin) and the reproducibility of detection is shown at each concentration tested in Table 84Error! Reference source not found ..

Table 84. Reproducibility of FilmArray Pneumonia Panel Bacterial Bin Results on FilmArray 2.0 and FilmArray Torch

Grey shading indicates the expected bin results based on the analyte concentration and bold font indicates the bin with the greatest percentage of results at each concentration.

BioFire Diagnostics 510(k)

FilmArray Pneumonia Panel

59

| | Concentration
(log10 copies/mL) | % Replicates Reported in Each Bin Result | | | | | Total
Detected |
|---------------------------------------------------------------------------|------------------------------------|------------------------------------------|------------------|--------------------------------------------------|------------------|----------------------|-------------------|
| Analyte | | ≥10^7 | 10^6 | 10^5 | 10^4 | ND | |
| Acinetobacter baumannii
(NDM-1)
AR-BANK#0033 | 7.5 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 6.5 | 87/90
(96.7%) | 3/90
(3.3%) | - | - | - | 90/90
100% |
| | 5.5 | - | 82/90
(91.1%) | 8/90
(8.9%) | - | - | 90/90
100% |
| | 4.5 | - | 1/90
(1.1%) | 80/90
(88.9%) | 9/90
(10.0%) | - | 90/90
100% |
| | 3.5 | - | - | 1/90
(1.1%) | 74/90
(82.2%) | 15/90
(16.7%) | 75/90
83.3% |
| | 2.5 | - | - | - | 1/90
(1.1%) | 89/90
(98.9%) | 1/90
1.1% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| Enterobacter cloacae
(VIM)
AR-BANK#0154 | 7.0 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 6.0 | 4/90
(4.4%) | 86/90
(95.6%) | - | - | - | 90/90
100% |
| | 5.0 | - | 6/90
(6.7%) | 80/90
(88.9%) | - | 4/90
(4.4%) | 86/90
95.6% |
| | 4.0 | - | - | 6/90
(6.7%) | 83/90
(92.2%) | 1/90
(1.1%) | 89/90
98.9% |
| | 3.0 | - | - | 1/90
(1.1%) | 4/90
(4.4%) | 85/90
(94.4%) | 5/90
5.6% |
| | 2.0 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| Escherichia coli
(IMP)
GRE 1062016 | 7.0 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 6.0 | 7/90
(7.8%) | 82/90
(91.1%) | - | - | 1/90
(1.1%) | 89/90
98.9% |
| | 5.0 | - | 10/90
(11.1%) | 80/90
(88.9%) | - | - | 90/90
100% |
| | 4.0 | - | - | 12/90
(13.3%) | 77/90
(85.6%) | 1/90
(1.1%) | 89/90
98.9% |
| | 3.0 | - | - | 1/90
(1.1%) | 15/90
(16.7%) | 74/90
(82.2%) | 16/90
17.8% |
| | 2.0 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| Haemophilus
influenzae
ATCC 10211 | 7.0 | 89/90
(98.9%) | 1/90
(1.1%) | - | - | - | 90/90
100% |
| | 6.0 | 35/90
(48.9%) | 55/90
(61.1%) | - | - | - | 90/90
100% |
| | 5.0 | - | 49/90
(54.4%) | 40/90
(44.4%) | 1/90
(1.1%) | - | 90/90
100% |
| | 4.0 | - | - | 41/90
(45.6%) | 49/90
(54.4%) | - | 90/90
100% |
| | 3.0 | - | - | - | 42/90
(46.7%) | 48/90
(53.3%) | 42/90
46.7% |
| | 2.0 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| Klebsiella aerogenes
(Enterobacter aerogenes)
ATCC 13048 | 7.5 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 6.5 | 65/90
(72.2%) | 25/90
(27.8%) | - | - | - | 90/90
100% |
| | 5.5 | - | 52/90
(57.8%) | 38/90
(42.2%) | - | - | 90/90
100% |
| | 4.5 | - | - | 38/90
(42.2%) | 51/90
(56.7%) | 1/90
(1.1%) | 89/90
98.9% |
| | 3.5 | - | - | - | 33/90
(36.7%) | 57/90
(63.3%) | 33/90
36.7% |
| | 2.5 | - | - | - | 1/90
(1.1%) | 89/90
(98.9%) | 1/90
1.1% |
| | Concentration
(log10 copies/mL) | % Replicates Reported in Each Bin Result | | | | | Total
Detected |
| Analyte | | ≥10^7 | 10^6 | 10^5 | 10^4 | ND | |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| Klebsiella oxytoca
(CTX-M)
GRE 1254054 | 7.5 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 6.5 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 5.5 | 1/90
(1.1%) | 84/90
(93.3%) | 3/90
(3.3%) | - | 2/90
(2.2%) | 88/90
97.8% |
| | 4.5 | - | - | 89/90
(98.9%) | - | 1/90
(1.1%) | 89/90
98.9% |
| | 3.5 | - | - | - | 90/90
(100%) | - | 90/90
100% |
| | 2.5 | - | - | 1/90
(1.1%) | 1/90
(1.1%) | 88/90
(97.8%) | 2/90
2.2% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| | 7.0 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 6.0 | 12/90
(13.3%) | 78/90
(86.7%) | - | - | - | 90/90
100% |
| | 5.0 | - | 15/90
(16.7%) | 75/90
(83.3%) | - | - | 90/90
100% |
| Klebsiella pneumoniae
(KPC)
AR-BANK#0097 | 4.0 | - | - | 23/90
(25.6%) | 66/90
(73.3%) | 1/90
(1.1%) | 89/90
98.9% |
| | 3.0 | - | - | - | 15/90
(16.7%) | 75/90
(83.3%) | 15/90
16.7% |
| | 2.0 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| | 7.0 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 6.0 | 26/90
(28.9%) | 64/90
(71.1%) | - | - | - | 90/90
100% |
| | 5.0 | - | 6/90
(6.7%) | 83/90
(92.2%) | 1/90
(1.1%) | - | 90/90
100% |
| Moraxella catarrhalis
ATCC 8176 | 4.0 | - | - | 4/90
(4.4%) | 86/90
(95.6%) | - | 90/90
100% |
| | 3.0 | - | - | - | 4/90
(4.4%) | 86/90
(95.6%) | 4/90
4.4% |
| | 2.0 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| | 7.0 | 88/90
(97.8%) | - | - | - | 2/90
(2.2%) | 88/90
97.8% |
| | 6.0 | 27/90
(30.0%) | 63/90
(70.0%) | - | - | - | 90/90
100% |
| Proteus mirabilis
ATCC 35659 | 5.0 | - | 26/90
(28.9%) | 64/90
(71.1%) | - | - | 90/90
100% |
| | 4.0 | - | - | 14/90
(15.6%) | 75/90
(83.3%) | 1/90
(1.1%) | 89/90
98.9% |
| | 3.0 | - | - | - | 28/90
(31.1%) | 62/90
(68.9%) | 28/90
31.1% |
| | 2.0 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| | 7.0 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 6.0 | 20/90
(22.2%) | 70/90
(77.8%) | - | - | - | 90/90
100% |
| Pseudomonas aeruginosa
ATCC 10145 | 5.0 | - | 24/90
(26.7%) | 66/90
(73.3%) | - | - | 90/90
100% |
| | 4.0 | - | - | 16/90
(17.8%) | 74/90
(82.2%) | - | 90/90
100% |
| | 3.0 | - | - | - | 14/90
(15.6%) | 76/90
(84.4%) | 14/90
15.6% |
| | Concentration
(log10 copies/mL) | ≥10^7 | 10^6 | % Replicates Reported in Each Bin Result
10^5 | 10^4 | ND | Total
Detected |
| | 2.0 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| | 7.0 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 6.0 | 2/90
(2.2%) | 88/90
(97.8%) | - | - | - | 90/90
100% |
| | 5.0 | - | 7/90
(7.8%) | 83/90
(92.2%) | - | - | 90/90
100% |
| Serratia marcescens
(OXA-48-like)
GRE 1659005 | 4.0 | - | - | 6/90
(6.7%) | 83/90
(92.2%) | 1/90
(1.1%) | 89/90
98.9% |
| | 3.0 | - | - | 6/90
(6.7%) | 84/90
(93.3%) | - | 6/90
6.7% |
| | 2.0 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| | 7.0 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 6.0 | - | 90/90
(100%) | - | - | - | 90/90
100% |
| Staphylococcus aureus
subsp. aureus
(mecA/C and MREJ)
ATCC 43300 | 5.0 | - | - | 90/90
(100%) | - | - | 90/90
100% |
| | 4.0 | - | - | - | 89/90
(98.9%) | 1/90
(1.1%) | 89/90
98.9% |
| | 3.0 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | 2.0 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | None
(No Analyte) | - | - | - | 2/1260
(0.2%) | 1258/1260
(99.8%) | 2/1260
0.2% |
| | 7.8 | 89/90
(98.9%) | 1/90
(1.1%) | - | - | - | 90/90
100% |
| | 6.8 | 89/90
(98.9%) | - | - | - | 1/90
(1.1%) | 89/90
98.9% |
| | 5.8 | - | 88/90
(97.8%) | 1/90
(1.1%) | 1/90
(1.1%) | - | 90/90
100% |
| Streptococcus agalactiae
ATCC 13813 | 4.8 | - | - | 89/90
(98.9%) | 1/90
(1.1%) | - | 90/90
100% |
| | 3.8 | - | - | - | 86/90
(95.6%) | 4/90
(4.4%) | 86/90
95.6% |
| | 2.8 | - | - | - | 3/90
(3.3%) | 87/90
(96.7%) | 3/90
3.3% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| | 6.5 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 5.5 | - | 90/90
(100%) | - | - | - | 90/90
100% |
| | 4.5 | - | - | 89/90
(98.9%) | 1/90
(1.1%) | - | 90/90
100% |
| Streptococcus pneumoniae
ATCC 6303 | 3.5 | - | - | - | 89/90
(98.9%) | 1/90
(1.1%) | 89/90
98.9% |
| | 2.5 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | 1.5 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| Streptococcus pyogenes
ATCC 49399 | 7.8 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 6.8 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 5.8 | 5/90
(5.6%) | 84/90
(93.3%) | 1/90
(1.1%) | - | - | 90/90
100% |
| | 4.8 | - | 4/90
(4.4%) | 86/90
(95.6%) | - | - | 90/90
100% |
| Analyte | Concentration
(log10 copies/mL) | ≥10^7 | 10^6 | 10^5 | 10^4 | ND | Total
Detected |
| | 3.8 | - | - | 3/90
(3.3%) | 87/90
(96.7%) | - | 90/90
100% |
| | 2.8 | - | - | - | 16/90
(18.9%) | 74/90
(81.1%) | 16/90
17.8% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |

60

61

62

The precision of the antimicrobial resistance (AMR) genes was measured as the reproducibility of analyte detection on each system and overall, presented in Error! Reference source not found. as the percent of replicates that are detected at concentrations of the associated bacterium that are within the reportable range, or below the reportable range, as well as the percent with the expected Not Detected result in unspiked samples.

Table 85. Reproducibility of FilmArray Pneumonia Panel Antimicrobial Resistance Gene Results on FilmArray 2.0 and FilmArray Torch

| | Concentration of
Organism
(log10 copies/mL) | Expected
Result | Agreement with the Expected Result | | | All Systems/Sites
[95% CI] |
|--------------------------------------------------------|---------------------------------------------------|------------------------|------------------------------------|-------------------------|------------------------------|------------------------------------|
| AMR Gene
Organism | | | FilmArray | FilmArray 2.0 | FilmArray
Torch | |
| | | | Site A | Site B | Site C | |
| CTX-M
Klebsiella oxytoca
GRE 1254054 | Reportable Range
(3.5 - 7.5) | Detected | 150/150
100% | 149/150a
99.3% | 150/150
100% | 449/450a
99.8%
[98.8%-99.9%] |
| | Below Reportable
Range
(2.5) | Detected
(Variable) | 0/30
0.0% | 1/30
3.3% | 0/30
0.0% | 1/90
1.1%
[0.03%-6.0%] |
| | None
(No Analyte) | N/A or
Not Detected | 600/600
100% | 599/600
99.8% | 600/600
100% | 1799/1800
99.9%
[99.7%-100%] |
| IMP
Escherichia coli
GRE 1062016 | Reportable Range
(4.0 - 7.0) | Detected | 120/120
100% | 120/120
100% | 120/120
100% | 360/360
100%
[99.0%-100%] |
| | Below Reportable
Range
(2.0-3.0) | Detected
(Variable) | 10/60
16.7% | 9/60
15.0% | 3/60
5.0% | 22/180
12.2%
[7.8%-17.9%] |
| | None
(No Analyte) | N/A or
Not Detected | 600/600
100% | 600/600
100% | 600/600
100% | 1800/1800
100%
[99.8%-100%] |
| KPC
Klebsiella pneumoniae
AR-Bank#0097 | Reportable Range
(4.0 - 7.0) | Detected | 120/120
100% | 119/120b
99.2% | 120/120
100% | 359/360b
99.7%
[98.5%-100%] |
| | Below Reportable
Range
(2.0-3.0) | Detected
(Variable) | 14/60
23.3% | 12/60
20.0% | 9/60
15.0% | 35/180
19.4%
[13.9%-25.0%] |
| | None
(No Analyte) | N/A or
Not Detected | 600/600
100% | 600/600
100% | 600/600
100% | 1800/1800
100%
[99.8%-100%] |
| mecA/C and MREJ
Staphylococcus aureus
ATCC 43300 | Reportable Range
(4.0 - 7.0) | Detected | 119/120b
99.2% | 118/120b
98.3% | 120/120
100% | 357/360b
99.2%
[97.6%-99.8%] |
| | Below Reportable
Range
(2.0-3.0) | Detected
(Variable) | 0/60
0.0% | 0/60
0.0% | 0/60
0.0% | 0/180
0%
[0.0%-2.0%] |
| | None
(No Analyte) | N/A or
Not Detected | 420/420
100% | 420/420
100% | 420/420
100% | 1260/1260
100%
[99.7%-100%] |
| NDM
Acinetobacter
baumannii
AR-Bank#0033 | Reportable Range
(3.5 - 7.5) | Detected | 150/150
100% | 149/150a
99.3% | 150/150
100% | 449/450a
99.8%
[98.8%-100%] |
| | Below Reportable
Range
(2.5) | Detected
(Variable) | 1/30
3.3% | 1/30
3.3% | 0/30
0.0% | 2/90
2.2%
[0.3%-7.8%] |
| | None
(No Analyte) | N/A or
Not Detected | 599/600
99.8% | 600/600
100% | 600/600
100% | 1799/1800
99.9%
[99.7%-100%] |
| OXA-48-like Serratia | Reportable Range | Detected | 120/120 | 119/120b | 120/120 | 359/360b |
| AMR Gene
Organism | Concentration of
Organism
(log10 copies/mL) | Expected
Result | FilmArray
Site A | FilmArray 2.0
Site B | FilmArray
Torch
Site C | All Systems/Sites
[95% CI] |
| marcescens
GRE 1659005 | (4.0 - 7.0) | Detected | 100% | 99.2% | 100% | 99.70%
[98.5%-100%] |
| | Below Reportable
Range
(2.0 - 3.0) | Detected
(Variable) | 14/60
23.3% | 12/60
20.0% | 9/60
15.0% | 35/180
19.4%
[13.9%-26.0%] |
| | None
(No Analyte) | N/A or
Not Detected | 598/600
99.7% | 600/600
100% | 600/600
100% | 1798/1800
99.9%
[99.6%-100%] |
| VIM
Enterobacter cloacae
AR-BANK#0154 | Reportable Range
(4.0 - 7.0) | Detected | 120/120
100% | 120/120
100% | 120/120
100% | 360/360
100%
[99.0%-100%] |
| | Below Reportable
Range
(2.0-3.0) | Detected
(Variable) | 10/60
16.7% | 9/60
15.0% | 3/60
5.0% | 22/180
12.2%
[7.8%-17.9%] |
| | None
(No Analyte) | N/A or
Not Detected | 599/600
99.8% | 600/600
100% | 600/600
100% | 1799/1800
99.9%
[99.7%-100%] |

63

3 CTX-M and NDM Not Detected results observed at the corresponding bacterial concentration of 4.5 logio copies/mL.

b KPC, mecA/C and MREI, and OXA-48-like Not Detected results observed at the corresponding bacterial of 4.0 log10 copies/mL.

Interference

Potentially interfering substances that could be present in BAL-like or sputum-like specimens or that may be introduced during specimen collection and testing were evaluated for their effect on FilmArray Pneumonia Panel performance. Substances included endogenous substances that may be found in specimens at normal or elevated levels (e.g. blood, mucus/mucin, human genomic DNA), various commensal or infectious microorganisms, medications, a variety of sample processing substances and substances used to clean, decontaminate, or disinfect work areas. The performance of the FilmArray Pneumonia Panel has not been established with all potentially interfering medications for the treatment of lower respiratory tract infections. The effect of interfering substances has only been evaluated for those listed in Table 86. Interference from substances that were not evaluated could lead to erroneous results.

Each substance was added to contrived samples containing representative qualitatively reported organisms and representative organisms with bin reporting. Qualitatively reported organisms were at concentrations near (2-3×) LoD and those with bin reporting were present at 4.0 log10 (copies/mL) (e.g. in the lowest reported bin). The concentration of substance added to the samples (Table 86) was equal to or greater than the highest level expected to be in BAL-like or Sputum-like specimens.

Four of the evaluated substances were found to interfere with the FilmArray Pneumonia Panel to report accurate analyte results; Bleach, MycoPrep, 2% NaOH, and 5% Oxalic acid. Each of these substances contain chemicals known to react with nucleic acids, altering their chemical structure. The interference observed was related to the inability to detect the chemically modified nucleic acids. Treatment of specimens with these substances prior to FilmArray Pneumonia Panel testing may result in loss of analyte detection, therefore samples that have been in contact with these substances should not be tested using the FilmArray Pneumonia Panel. None of the other substances tested were shown to interfere with the FilmArray Pneumonia Panel results, however, testing of specimens that have been centrifuged or pre-treated by addition of enzyme, media, mucolytic agent, or decontaminating substances is not recommended..

64

SubstanceConcentration TestedTesting Outcome
Endogenous Substances
Blood10% v/vNo Interference
Albumin60 mg/mLNo Interference
HCl (gastric acid)5 mmol/LNo Interference
Hemoglobin2 mg/mLNo Interference
Human Cells (K-562 cell line)3.8E+06 cells/mLNo Interference
Immunoglobulins (IgG)60 mg/mLNo Interference
Mucin16 mg/mLNo Interference
Exogenous Substances
Albuterol (bronchodilator)1.7 µmol/LNo Interference
Benzocaine (Orajel)1.0 % w/vNo Interference
Epinephrine (hormone, bronchodilator)8.3 µg/mLNo Interference
Galphimia glauca (Homeopathic remedy)1.0 % w/vNo Interference
Guaifenesin (expectorant)15.2 mmol/LNo Interference
Lidocaine5.1 mmol/LNo Interference
Menthol and cetylpyridinium chloride
(Cepacol Mouthwash)1.0% v/vNo Interference
Mupirocin (antibiotic)6.0 ng/mLNo Interference
Nicotine6.2 µmol/LNo Interference
Pentamidine (antimicrobial)1.5 mg/mLNo Interference
Phenylephrine hydrochloride (decongestant)0.3 mg/mLNo Interference
Tobramycin sulfate (antibiotic)30 mg/mLNo Interference
Zanamivir (influenza antiviral)426 ng/mLNo Interference
Competitive Microorganisms
Actinobacillus actinomycetecomitans3.8E+07 CFU/mLNo Interference
Aspergillus fumigatus5.5E+07 CFU/mLNo Interference
Burkholderia cepacia1.7E+07 CFU/mLNo Interference
Cryptococcus neoformans2.5E+05 CFU/mLNo Interference
Enterovirus D681.4E+06 copies/mLNo Interference
Haemophilus influenzae1.8E+07 CFU/mLNo Interference
Legionella pneumophila8.1E+06 CFU/mLNo Interference
Respiratory Syncytial Virus3.5E+04 copies/mLNo Interference
Staphylococcus epidermidis1.9E+07 CFU/mLNo Interference
Streptococcus mutans5.9E+06 CFU/mLNo Interference
Streptococcus pyogenes5.5E+06 CFU/mLNo Interference
Varicella Zoster Virus8.7E+07copies/mLNo Interference
Disinfection/Cleaning Substances
Reagent Alcohol7.0%No Interference
Bleach1.0% v/v
(600 ppm chlorine)Interference Observed a
Sample Processing Materialsa
Copan Snotbuster (active ingredient DTT)50.0% v/vNo Interference
Sputolysin (active ingredient DTT)50.0% v/vNo Interference
SPUTASOL (active ingredient DTT + salts)50.0% v/vNo Interference
MycoPrep (active ingredient NaOH + NALC)50.0% v/vInterference Observed b
NaOH (decontaminant)1.0%Interference Observed b
Oxalic Acid (decontaminant)2.5%Interference Observed b

Table 86. Evaluation of Potentially Interfering Substances on the FilmArray Pneumonia Panel

ª FilmArray Pneumonia Panel testing of lower respiratory specimens that have been processed or treated with these or other substances (e.g. trypsin) has not been validated and is not recommended.

b Pouch controls passed but Not Detected results were reported for one or more analytes after incubation of the sample with substance. Substance(s) are known to chemically interact with and damage nucleic acids (DNA and/or RNA) to prevent amplification.