The BIOFIRE FILMARRAY Pneumonia Panel (BIOFIRE Pneumonia Panel) is a multiplexed nucleic acid test intended for use with BIOFIRE FILMARRAY 2.0 (BIOFIRE 2.0) or BIOFIRE FILMARRAY TORCH (BIOFIRE TORCH) systems for the simultaneous detection of multiple respiratory viral and bacterial nucleic acids, as well as select antimicrobial resistance genes, in sputum-like speciorated sputum, or endotracheal aspirates) or bronchoalveolar lavage (BAL)-like specimens (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, 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
• · Klebsiella oxytoca
• · Serratia marcescens
• · Enterobacter cloacae complex
• Klebsiella pneumoniae group
• · Staphylococcus aureus
• · Escherichia coli
• · Moraxella catarrhalis
• · Streptococcus agalactiae
• Haemophilus influenzae
• · Proteus spp.
• · Streptococcus pneumoniae
• Klebsiella aerogenes
• Pseudomonas aeruginosa
• · Streptococcus pyogenes

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

Atypical Bacteria

• Chlamydia pneumoniae
• · Legionella pneumophila
• Mycoplasma pneumoniae

Viruses

• · Adenovirus
• Human rhinovirus/enterovirus
• · Parainfluenza virus
• · Coronavirus
• · Influenza A virus
• Respiratory syncytial virus

• Human metapneumovirus

• Influenza B virus
  Antimicrobial Resistance Genes

• · CTX-M

• IMP

• КРС

• NDM

• OXA-48-like

• VIM

• · mecA/C and MREJ (MRSA)

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 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 BIOFIRE 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 BIOFIRE 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 BIOFIRE 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 BIOFIRE 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.

BIOFIRE FILMARRAY Pneumonia Panel plus:

The BIOFIRE FILMARRAY Pneumonia Panel plus (BIOFIRE Pneumonia Panel plus) is a multiplexed nucleic acid test intended for use with BIOFIRE FILMARRAY 2.0 (BIOFIRE 2.0) or BIOFIRE FILMARRAY TORCH (BIOFIRE TORCH) systems for the simultaneous detection and identification of nucleic acids from Middle East respiratory syndrome coronavirus (MERS-CoV) and 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 meeting MERS-CoV clinical and/or epidemiological criteria.

Testing with BIOFIRE Pneumonia Panel plus should not be performed unless the patient meets clinical and/or epidemiologic criteria for testing suspected MERS-CoV specimens. Thical signs and symptoms assocated with MERS-CoV infection, contact with a probable or confirmed MERS-CoV case, history of travel to geographic locations where MERS-CoV cases were detected, or other epidemiological links for which MERS-CoV testing may be indicated.

The following bacteria are reported semi-quantitatively with bins representing approximately 10^4, 10^5, 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

• · Middle East respiratory syndrome coronavirus (MERS-CoV)
• Adenovirus
• Coronavirus
• Human metapneumovirus
• Human rhinovirus/enterovirus
• · Influenza A virus
• Influenza B virus
• Parainfluenza virus
• · Respiratory syncytial virus

Antimicrobial Resistance Genes

• CTX-M

• IMP

• · KPC

• NDM

• OXA-48-like

• VIM

• · mecA/C and MREJ (MRSA)

The detection and identification of specific viral and bacterial nucleic acids from MERS-CoV and other respiratory pathogens, as well as the estimation of relative abundance of nucleic acid from common bacterial analytes, within specimens collected from individuals meeting MERS-CoV clinical and/or epidemiological criteria aids in the differential diagnosis of MERS-CoV infection, if used in conjunction with other clinical and epidemiological information in accordance with the guidelines provided by the appropriate public health authorities.

BIOFIRE Pneumonia Panel plus MERS-CoV positive results are for the presumptive identification of MERS-CoV. The definitive identification of MERS-CoV requires additional testing and confirmation procedures in consultation with the appropriate public health authorities (e.g., local or state public health departments, etc.) for whom reporting is necessary. The diagnosis of MERS-CoV infection must be made based on history, signs, symptoms, exposure likelihood, and other laboratory evidence in addition to the identification of MERS-CoV.

BIOFIRE Pneumonia Panel plus MERS-CoV negative results, even in the context of a BIOFIRE Pneumonia Panel plus positive result for one or more of the common respiratory pathogens, do not preclude MERS-CoV infection and should not be used as the sole basis for patient management decisions. The levels of MERS-CoV that would be present in sputum-like or BAL-like specimens from individuals with early infection and from asymptomatic MERS-CoV carriers are not well understood. A negative BIOFIRE Pneumonia Panel plus MERS-CoV result in an asymptomatic individual does not rule out the possibility of future illness and does not demonstrate that the individual is not infectious.

Viral culture should not be attempted on specimens with positive BIOFIRE Pneumonia Panel plus results for MERS-CoV unless a BSL 3 facility is available to receive and culture specimens.

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 BIOFIRE Pneumonia Panel plus 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 BIOFIRE Pneumonia Panel plus 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 BIOFIRE Pneumonia Panel plus 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 BIOFIRE Pneumonia Panel plus, 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.

The BIOFIRE® FILMARRAY® Pneumonia Panel and BIOFIRE® FILMARRAY® Pneumonia Panel plus use nested, multiplex reverse transcription polymerase chain reaction (PCR), followed by melting curve analysis for the detection of select organisms and antimicrobial resistance (AMR) genes in sputum-like (induced and expectorated sputum as well as endotracheal aspirate, ETA) and bronchoalveolar lavage (BAL)-like (BAL and mini-BAL) specimens. The panels allow for the identification of specific bacteria, atypical bacteria, viruses, and AMR genes as indicated in Table 1. The BIOFIRE Pneumonia Panel and BIOFIRE Pneumonia Panel plus pouches are identical, but the BIOFIRE Pneumonia Panel plus includes reporting of Middle East Respiratory Syndrome Coronavirus (MERS-CoV), which is not included in the BIOFIRE Pneumonia Panel. Reporting of MERS-CoV is controlled through software masking of the MERS-CoV result for the BIOFIRE Pneumonia Panel.

The BIOFIRE Pneumonia Panels are compatible with bioMérieux's PCR-based in vitro diagnostic BIOFIRE® FILMARRAY® 2.0 and BIOFIRE® FILMARRAY® TORCH Systems for infectious disease testing. Specific software module (i.e. BIOFIRE Pneumonia Panel Pouch Module Software) are used to perform BIOFIRE Pneumonia Panels testing on these systems.

This document refers to a 510(k) premarket notification for a medical device (BIOFIRE FILMARRAY Pneumonia Panel and BIOFIRE FILMARRAY Pneumonia Panel plus). This type of submission focuses on demonstrating substantial equivalence to a legally marketed predicate device rather than presenting a full de novo study with strict acceptance criteria and performance validation against a test set. The document clearly states that the submission is for software updates to mitigate false positive Coronavirus and CTX-M results and that "Reanalysis of the performance data with the modified pouch module software did not result in an overall change of the study conclusions or performance claims for non-clinical/analytical studies."

Therefore, the information typically requested in your prompt regarding acceptance criteria, study details, sample sizes, expert ground truth establishment, MRMC studies, and standalone performance might not be explicitly detailed in this type of FDA submission as it would be for a de novo marketing authorization. However, I can extract what is available and clarify what is not.

Based on the provided text, here's a breakdown of the requested information:

1. A table of acceptance criteria and the reported device performance

The document does not explicitly state formal "acceptance criteria" in a quantitative table format as might be seen for a new device submission. Instead, the focus is on the impact of the software update on existing performance. The relevant performance change mentioned is:

The document implies that these updated specificities are acceptable because they represent an improvement in mitigating false positives and "did not result in an overall change of the study conclusions or performance claims for non-clinical/analytical studies."

2. Sample sizes used for the test set and the data provenance (e.g. country of origin of the data, retrospective or prospective)

The document mentions a "Clinical Prospective Study" for which the Coronavirus specificity numbers are reported. It does not provide the exact sample size for this specific study, nor does it explicitly state the country of origin. It indicates that the reanalysis of existing performance data was done.

3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts (e.g. radiologist with 10 years of experience)

This information is not provided in the document. As this is a molecular diagnostic test, ground truth would typically be established by highly sensitive and specific laboratory methods (e.g., PCR, sequencing, culture) rather than expert human interpretation in the way radiologists interpret images.

4. Adjudication method (e.g. 2+1, 3+1, none) for the test set

This information is not provided.

5. If a multi-reader multi-case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance

An MRMC study is not applicable here as this is a molecular diagnostic device, not an AI-assisted diagnostic imaging device that involves human reader interpretation.

6. If a standalone (i.e. algorithm only without human-in-the-loop performance) was done

This refers to the performance of the assay itself. The document implicitly discusses the "standalone" performance of the BIOFIRE FILMARRAY Pneumonia Panel and Panel Plus, which is a molecular diagnostic test. The reported specificities are a measure of this standalone performance. The software update is an internal modification to the assay's interpretation logic.

7. The type of ground truth used (expert consensus, pathology, outcomes data, etc)

The document implies that the ground truth for the clinical performance evaluations was established through highly sensitive and specific methods for pathogen detection, as is standard clinical laboratory practice for molecular diagnostics. It does not explicitly state the specific ground truth methods but mentions that "Culture is required to obtain isolates for antimicrobial susceptibility testing, and BIOFIRE Pneumonia Panel results should be used in conjunction with culture results for determination of bacterial susceptibility or resistance." This suggests that culture and other definitive laboratory tests would be part of the ground truth establishment, particularly for bacterial analytes and antimicrobial resistance genes.

8. The sample size for the training set

This document describes a software update to an already cleared device. It does not provide details about a "training set" in the context of machine learning model development. The software update appears to be a rule-based or algorithmic adjustment to optimize melting curve analysis and mitigate cross-reactivity with human genomic DNA, rather than a re-training of a complex AI model. The modification was driven by "routine post-market monitoring and complaint investigations combined with concurrent findings from an internal product development study."

9. How the ground truth for the training set was established

As there's no mention of a traditional "training set" in the context of an AI model, this information is not provided. The "ground truth" that informed the software change was likely observations of false positives in clinical samples, identified through investigations and potentially confirmed by orthogonal testing or characterization of the offending interactions (cross-reactivity with hgDNA).

Ask a specific question about this device

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 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 -KPC -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 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 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 CFUmL. 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.

The FilmArray Pneumonia (PN) Panel is designed to simultaneously identify 26 potential pathogens of lower respiratory tract infection (LRTI) and associated antimicrobial resistance (AMR) genes from a sputum-like (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 PN Panel is compatible with BioFire Diagnostics' (BioFire) PCR-based in vitro diagnostic FilmArray 2.0 (K143178) and FilmArray Torch (K160068) systems for infectious disease testing. A specific software module (i.e. FilmArray PN Panel pouch module) is used to perform FilmArray PN 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
Bacteria (Atypical) - Qualitative Results: Chlamydia pneumoniae, Legionella pneumophila, Mycoplasma pneumoniae
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)
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 PN 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 pouch into 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 first stage, 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 single plex 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.

A feature of the FilmArray PN Panel is the reporting of organism abundance for common bacteria in discrete bins representing 10^4, 10^5, 10^6, and >10^7 genomic copies/mL. The panel accomplishes this by comparing the amplification of the bacterial assays with that of a Quantified Standard Material (QSM) present in the pouch.

Here's an analysis of the provided text regarding the FilmArray Pneumonia Panel, focusing on the acceptance criteria and study details. It's important to note that this document is a Special 510(k) Summary for a labeling modification related to a previously cleared device. Therefore, it primarily discusses the change and its impact on the device's labeling, rather than presenting a comprehensive de novo validation study.

Key takeaway: The document describes a labeling modification due to a stability issue with the Adenovirus2 assay (specifically for Adenovirus C) within 6 months of the pouch expiration date. It does not provide details of an initial, full validation study with acceptance criteria and reported performance for all analytes, as that would have been part of the original K180966 submission. The information below is extracted from what's available in this specific document regarding the impact of the noted issue.

1. Table of Acceptance Criteria and Reported Device Performance

This document does not present a table of general acceptance criteria and reported performance for all analytes of the FilmArray Pneumonia Panel since it's a labeling modification submission. The focus is specifically on the change in performance for Adenovirus C due to stability.

The "acceptance criteria" discussed here are essentially the observed degradation in sensitivity and the resulting limitation on the use of the device for Adenovirus C detection under specific conditions.

Note: The original acceptance criteria for the initial clearance (K180966) would have included specific sensitivity (Limit of Detection - LoD) and specificity targets, which are not detailed in this specific document.

2. Sample Size Used for the Test Set and Data Provenance

The document does not specify a "test set" in the context of a de novo validation study. Instead, it refers to a stability study where the issue with Adenovirus2 assay's performance was discovered.

• Sample Size for Test Set: Not explicitly stated as a separate test set. The issue was identified during a stability study of the device pouches.
• Data Provenance: The issue was identified through internal stability study results conducted by BioFire Diagnostics, LLC. No country of origin for clinical samples is mentioned, as the data appears to be from analytical testing (stability of laboratory-manufactured pouches). The study type is retrospective in the sense that previously manufactured pouches were being tested for stability over time.

3. Number of Experts Used to Establish Ground Truth and Qualifications

Not applicable. This document does not describe a study involving expert consensus to establish ground truth for clinical cases. The issue identified was an analytical performance degradation discovered during internal stability testing.

4. Adjudication Method

Not applicable. This document is not describing a study that required adjudication of complex clinical cases or image interpretations. The "adjudication" was the internal assessment of stability study results and the determination that the performance characteristic (LoD for Adenovirus C) had degraded under specific conditions.

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study

Not applicable. The device is an in vitro diagnostic (IVD) based on molecular detection, not one requiring human interpretation of results. Therefore, an MRMC study is not relevant to this type of device.

6. Standalone (Algorithm Only) Performance

Yes, the information presented relates to standalone performance of the device. The reported impairment in LoD for Adenovirus C was determined through analytical testing of the device itself (the assay in the pouch) under specific storage conditions. There is no human-in-the-loop component in the detection process of the FilmArray Pneumonia Panel.

7. Type of Ground Truth Used

The ground truth used for identifying this issue was analytical performance data (stability study results) and the degradation of the Limit of Detection (LoD) for Adenovirus C, which represents a quantifiable measure of the assay's sensitivity. This is akin to a "spike-in" experiment or testing known positive controls at various concentrations across the shelf-life of the product.

8. Sample Size for the Training Set

Not applicable. This document describes a stability issue with an already-cleared device and a subsequent labeling modification. It does not refer to a "training set" in the context of an algorithm or AI development. The device pre-dates common AI/ML nomenclature in medical device submissions for IVDs.

9. How the Ground Truth for the Training Set Was Established

Not applicable, as there is no "training set" described in this document.

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The Unyvero LRT BAL Application is a qualitative nucleic acid multiplex test intended for the simultaneous detection and identification of nucleic acid sequences from the following microorganisms (N = 20) and antibiotic resistance markers (N = 10) in bronchoalveolar lavage (BAL)-like specimens (BAL or mini-BAL) from adult hospitalized patients with suspected lower respiratory tract infections.

The Unyvero LRT BAL Application performed on the Unyvero System is indicated as an aid in the diagnosis of lower respiratory tract infection in adult hospitalized patients with signs and symptoms of lower respiratory infection; results should be used in conjunction with other clinical and laboratory findings. As BAL specimens may contain colonizing microorganisms, detection of Unyvero LRT BAL microbial targets does not indicate that the microorganism is the disease. Unyvero positive results do not rule out co-infection with other microorganisms. Negative results do not preclude lower respiratory infection, as the causative agent may be a microorganism not detected by this test.

A negative result for any antibiotic resistance marker does not indicate that detected microorganisms are susceptible to applicable antimicrobial agents. Detected antibiotic resistance markers cannot be definitively linked to specific microorganisms, and may be present in organisms that are not detected by the Unyvero LRT BAL Application.

Microbiology cultures of BALs should be performed to obtain isolates for species identification and antimicrobial susceptibility testing and to identify potential microorganisms not targeted by the Unyvero LRT BAL Application.

The Unyvero LRT BAL Application automates and integrates DNA purification and eight parallel multiplex endpoint PCR reactions. It provides qualitative detection of nucleic acids from multiple lower respiratory pathogens using hybridization on PCR chamber arrays in a single use cartridge from a single bronchoalveolar lavage (BAL)-like specimen (BAL or mini-BAL).

The Unyvero LRT BAL Application identifies 20 microorganisms and 10 antibiotic resistance markers as listed in the Intended Use Statement, below.

The Unyvero LRT BAL Application uses a multiplex PCR approach following array hybridization which targets 30 individual analytes (microorganisms (N = 20) and antibiotic resistance markers (N = 10) divided into eight separate PCR reactions that are performed in individual reaction chambers simultaneously on a Unyvero LRT BAL Application cartridge. Multiplex compositions are designed to avoid any expected common occurrence of certain analytes within the same multiplex to largely reduce competitive PCR inhibition. Individual analyte assays of the Unyvero LRT BAL panel are designed to exhibit low or absent cross-reactivity with the relevant bronchoalveolar lavage (BAL)like specimens (BAL or mini-BAL) sample matrix or 'close neighbor' strains. Array oligonucleotides are designed for similar hybridization and melting temperatures (approx. 65 - 80 ℃, varying by amplicon). Hybridization and melting temperatures are used to exclude non-specific hybridization signals for improved signal specificity.

The instrumentation consists of one (or more) Unyvero L4 Lysator, one (or more) Unyvero A50 Analyzer, a Unyvero C8 Cockpit, and four single-use consumables: the Unyvero LRT BAL Cartridge, the Unyvero Sample Tube, Sample Tube Cap and the Unyvero Master Mix. A Unyvero Sample Tube Holder is supplied as accessory to simplify the sample filling step.

• Unyvero LRT BAL Cartridge contains DNA isolation and purification reagents, . primers, hybridization and wash buffers, and oligonucleotides for detection.
• Unyvero T1 Sample Tube and Transport Cap contains glass beads and buffers to lyse ● microorganisms and liquefy the sample.
• . Unyvero T1 Sample Tube Cap seals the Unyvero Sample Tube and contains Proteinase K and a synthetic control gene for process monitoring.
• Unyvero M1 Master Mix Tube contains reagents for DNA amplification. ●

An internal control (a synthetic gene without any homology to known sequences) is processed in every chamber in order to verify the DNA purification, array hybridization, and detection.

Other than the built-in controls, no external materials are supplied with Unyvero LRT BAL Application devices and consumables. Good laboratory practice recommends running external positive and negative controls using samples cultured in the laboratory.

No additional reagents are required to perform the Unyvero LRT BAL Application; all reagents are supplied within the cartridge or within the other consumables with the exception of the polymerase Master Mix, which is provided separately (frozen).

How to perform a Unyvero LRT BAL test:

• 1. Remove the Unyvero Sample Tube from its packaging and slide it in the Unyvero Sample Tube Holder in the upright position with the barcoded end at the bottom.
• 2. Remove the Transport Cap from the Sample Tube by pulling it upward.
• 3. Pipette 180 uL of vortexed patient specimen into the Sample Tube and close it using the Unyvero Cap provided in the LRT BAL kit: align the small nodules on the neck of the Sample Tube with the Tube with the openings on the Cap and press down to lock in place.
• 4. Scan the Sample Tube and place it into the Lysator. Close the Lysator lid to start the Lysator.
• 5. Remove Master Mix from freezer and thaw at room temperature (15 ℃ 25 ℃) for approximately 30 minutes.
• 6. When lysis is complete, remove the Sample Tube from the Lysator and place it into the labeled position on the left-hand side of the Unyvero LRT BAL Cartridge.
• 7. Place the thawed Master Mix into the labeled position on the right-hand side of the Cartridge.
• 8. Scan the Cartridge on the Cockpit and insert it into the position indicated on the Analyzer. The software provides on-screen instructions to start the test.
• 9. View results after completion of the run.

During the automated analysis (Step 8), which is entirely controlled by the A50 Analyzer, the sample is mixed with ethanol and then transferred onto the DNA purification column, where buffers purify and elute the DNA. Eluted DNA is transferred to a chamber, where mixing with the Master Mix takes place. This mixture is distributed into eight separate PCR reaction chambers each containing multiple primer pairs, consisting of one labeled and one non-labeled primer for the respective multiplex endpoint PCR. After specific amplification, PCR products are hybridized to the corresponding array probes. Each array has been manufactured with probes corresponding to the amplicons for the targeted microorganism sequences described above. A total of up to 49 spots per array allows for redundant detection with at least four spots per analyte, as well as spots for intensity calibration, and orientation markers for the image processing software. Binding of amplicons to specific probes is detected by analyzing fluorescence images of the arrays. Result data are displayed on the C8 Cockpit for visualization, printout, temporary storage and electronic data export.

A test run is completed after 4 to 5 hrs, and results for panel microorganisms and corresponding antibiotic resistance markers are displayed on the Cockpit screen. Four screens are provided:

• . The Result Summary screen provides a quick overview of all detected LRT BAL panel microorganisms, together with all detected corresponding antibiotic resistance markers.
• . The Microorganisms screen provides a list of all panel microorganisms grouped in Grampositive bacteria, non-fermenting bacteria, Enterobacteriaceae and other microorganisms together with the analysis result (reported as detected, not detected or invalid).
• The Result Details screen provides a list of all analyzed microorganisms and the ● corresponding antibiotic resistance markers together with the analysis result (reported as detected, not detected, invalid or NA).
• Test Details screen showing user name, lot numbers of used consumables, expiration dates of ● the consumables and start and stop times and dates of the test.

Results can be reviewed on the cockpit or, optionally, be printed out. All results are saved in a database on the Unyvero Cockpit for later review and printing.

The Unyvero software is designed to:

• Manage analysis workflow (Cockpit) ●
• Carry out sample lysis (Lysator) ●
• Execute the analysis and generate the analytical result (Analyzer) .
• Manage communication among units (Cockpit, Lysator, Analyzer) ●
• Monitor internal mechanical / electrical actuators (Lysator, Analyzer) ●
• Present analysis results (Cockpit) .
• Store analysis results (Cockpit) ●

Each device (Cockpit, Lysator. Analyzer) is a subsystem within the overall system, and each consists of hardware and software components. The different devices are interconnected by an Ethernet based communication interface, and system functionality is provided by the interaction of all three device types. Only the Cockpit presents a rich user interface and allows interaction with the operator. The Lysator and Analyzer units include a simple display for showing device status. Optional HIS/LIS connectivity allows transferring results to a hospital or laboratory information system.

This looks like a medical device submission, specifically a 510(k) summary for the "Unyvero LRT BAL Application." Let's break down the acceptance criteria and study data provided.

Acceptance Criteria and Device Performance (Summary derived from the provided text)

The document doesn't explicitly state "acceptance criteria" numerical thresholds in a single, consolidated table. However, regulatory submissions often imply acceptance criteria based on the demonstrated performance that is deemed sufficient for clearance. Based on the performance tables provided, particularly the "Comparison to Composite Comparator Reference" (Table 28), we can infer the device's demonstrated performance for each microorganism. For antibiotic resistance markers, the "Comparison of antibiotic resistance markers to corresponding molecular reference assays" (Table 29) shows reported performance.

Inferred Acceptance Criteria (Implicit from demonstrated performance):

• For Microorganism Detection (PPA & NPA values compared to a Composite Comparator): The FDA generally expects high Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA) to ensure the device is effective and safe. While no explicit thresholds are given, the achieved percentages, predominantly in the range of 80-100% for PPA and 90-100% for NPA across various microorganisms, suggest these levels were acceptable to the FDA.
• For Antibiotic Resistance Marker Detection (PPA & NPA values compared to Molecular Reference): Similar high PPA and NPA values were demonstrated (e.g., 88.9-100% for PPA and 69.5-100% for NPA), indicating these ranges were likely acceptable.
• Reproducibility: High agreement rates (over 90% for most cases, 100% for negative samples) for both moderate and low concentrations.
• Inclusivity & Exclusivity: Detection of target strains at or near LoD, and no cross-reactivity with non-target or common flora organisms.
• Limit of Detection (LoD): Demonstrated specific LoD values for each target (as listed in Tables 2 and 3).

Reported Device Performance (from Tables 28 & 29 for clinical accuracy):

Antibiotic Resistance Markers (Table 29):

Study Details

Here's a breakdown of the information regarding the studies:

1. Sample size used for the test set and the data provenance:

   • Prospective Study: 1,016 bronchoalveolar lavage (BAL) or mini-BAL specimens from hospitalized adult patients with suspected lower respiratory tract infections. Data provenance: Collected at nine US clinical sites between June 2015 and July 2016 for a previous study, stored frozen. Retrospective collection of prospectively collected samples.
   • Archived Study: 392 lavage specimens (BAL or mini-BAL) from patients (age 18 or older) with suspected lower respiratory infection and at least one positive LRT BAL panel microorganism by standard-of-care (SoC). Data provenance: Collected at 11 US clinical sites between 2015 and 2019, complemented by a few specimens from other sites. This included 197 specimens from a previous study and 195 new specimens. Retrospective collection.
   • Contrived Study: Used to supplement rare analytes. Consisted of 60 contrived specimens per target analyte (total not explicitly stated for all analytes combined, but implies 60 for each of the 13 microorganisms and relevant antibiotic resistance markers used in this component). Prepared by spiking pooled microorganisms into negative lavage specimens.

2. Number of experts used to establish the ground truth for the test set and the qualifications of those experts:

   • The document describes "Standard-of-Care (SoC) culture results" as one reference method for microorganisms and "molecular multiplex PCR comparator assay for which all positive PCR results were followed by bi-directional sequencing" as another.
   • For "atypical microorganisms," a combination of two different validated PCR comparator assays (each targeting different genetic loci) was used.
   • For antibiotic resistance markers, "PCR assays corresponding to each LRT BAL antibiotic resistance marker assay were included into the multiplex PCR comparator assay as a (single) molecular reference. Positive PCRs were followed by bi-directional sequencing."
   • "Cultured isolates had been collected for positive prospective specimens whenever possible. Isolates were regrown and evaluated by MALDI-TOF to confirm strain identities and by whole genome sequencing using a next generation sequencing (NGS) approach to screen for presence or absence of LRT BAL panel antibiotic resistance markers."
   • "Phenotypic AST results for positive specimens as reported by SoC culture were collected to correlate antibiotic resistance markers detected by the LRT BAL Application to resistance phenotypes."
   • The document does NOT explicitly mention the number or specific qualifications of "experts" (e.g., medical doctors, microbiologists, or other specialists) routinely involved in establishing the ground truth measurements (like interpreting SoC culture results, performing PCR/sequencing or AST). It refers to "standard-of-care" clinical laboratory procedures and "molecular reference assays," implying trained laboratory personnel, but no explicit "expert panel" or count is given as typically seen for imaging studies.

3. Adjudication method (e.g., 2+1, 3+1, none) for the test set:

   • For microbiology results: Discrepant results for false positives were analyzed by performing "singleplex PCRs/bi-directional sequencing using primer pairs targeting different genetic loci compared to the corresponding LRT BAL assays on specimen DNA extracts for each discrepant LRT BAL analyte." This indicates a form of molecular adjudication for discrepancies, rather than a consensus by human experts in the traditional sense, but still a specific method for resolving differences.
   • For antibiotic resistance markers, "Correlation of Detected Antibiotic Resistance Markers to Strain Genotypes and Phenotypes" involved comparing the device's results to both molecular sequencing of isolates and phenotypic antimicrobial susceptibility testing (AST) results. This also serves as a multi-modal "adjudication" or reference verification process.
   • No explicit "2+1" or "3+1" human reader adjudication method, common in imaging studies, is mentioned here, as it's a diagnostic test based on molecular/culture evidence.

4. If a multi-reader multi-case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance:

   • No, this is not applicable. The Unyvero LRT BAL Application is a qualitative nucleic acid multiplex test for microorganism and resistance marker detection. It is a standalone diagnostic device, not an AI-assisted interpretation tool for human readers (like radiologists interpreting images). Therefore, an MRMC comparative effectiveness study involving human reader improvement with/without AI is not relevant to this device.

5. If a standalone (i.e., algorithm only without human-in-the-loop performance) was done:

   • Yes, this was a standalone performance study. The device itself performs automated DNA purification, multiplex PCR, hybridization, and interpretation. The clinical performance data presented (PPA, NPA, etc.) represent the performance of the "Unyvero LRT BAL Application" as a full system, comparing its direct output to the established reference methods (SoC culture, molecular comparator assays, sequencing). There is no "human-in-the-loop" aspect to the test's result generation.

6. The type of ground truth used (expert consensus, pathology, outcomes data, etc.):

   • For "typical" microorganisms: The primary ground truth for the prospective study was Standard-of-Care (SoC) culture results, supplemented by a composite comparator (SoC culture combined with molecular multiplex PCR and bi-directional sequencing) to address limitations of culture.
   • For "atypical" microorganisms: The primary ground truth was a composite comparator consisting of two different validated PCR comparator assays (each with bi-directional sequencing).
   • For antibiotic resistance markers: Ground truth involved comparison to molecular reference assays (PCR/sequencing) for the specific marker and phenotypic antimicrobial susceptibility testing (AST) results for cultured isolates.

7. The sample size for the training set:

   • The document describes performance studies (prospective, archived, contrived) used for validation. It does NOT specify a distinct "training set" or its size. This is typical for a traditional (non-AI/machine learning) diagnostic device validation, where the focus is on a comprehensive validation dataset rather than a separate training and test split. The LoD, inclusivity, exclusivity, interference, and reproducibility studies use various laboratory-prepared samples.

8. How the ground truth for the training set was established:

   • Not applicable, as a specific "training set" is not mentioned in the context of this device. The validation studies utilized reference methods as described in point 6 above.

Ask a specific question about this device

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.

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.

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.

Acceptance Criteria and Device Performance for BioFire FilmArray Pneumonia Panel (K180966)

The BioFire FilmArray Pneumonia Panel is a multiplexed nucleic acid test intended for the simultaneous detection of multiple respiratory viral and bacterial nucleic acids, as well as select antimicrobial resistance genes, in respiratory specimens to aid in the diagnosis of lower respiratory tract infection.

1. Table of Acceptance Criteria and Reported Device Performance

The provided document does not explicitly state pre-defined acceptance criteria (e.g., "sensitivity must be >X%"). Instead, the performance is reported as the observed sensitivity/PPA and specificity/NPA from clinical and analytical studies, which are then used to demonstrate substantial equivalence to the predicate device.

Below is a summary of the reported performance for key analytes in the clinical study for BAL specimens, which served as a primary evaluation for the device's effectiveness. A similar detailed table for sputum is also available in the source document.

Summary of FilmArray Pneumonia Panel Clinical Performance (BAL Specimens)

2. Sample size used for the test set and the data provenance

Clinical Study (Test Set):

• Total specimens acquired: 904 residual BAL (821 BAL and 83 mini-BAL) and 925 residual sputum (478 sputum and 447 ETA).
• Final data set for analysis: 846 BAL and 836 sputum specimens after exclusions.
• Data provenance: Multi-center study conducted at eight geographically distinct U.S. study sites from October 2016 to July 2017. The study was prospective as it involved collecting specimens for testing.

Archived Specimens (Supplemental Test Set):

• Total specimens: 171 frozen archived specimens (13 BAL and 5 sputum negatives; 139 BAL and 14 sputum positives).
• Final data set for analysis: 18 negative and 149 positive specimens (containing 173 analytes).
• Data provenance: Retrospective, preselected archived specimens from external laboratories.

Contrived Specimens (Supplemental Test Set):

• Total specimens: 1125 contrived specimens (spiked using residual clinical samples).
• Data provenance: Generated in-house by BioFire (presumably in the US) using residual clinical samples that were pre-screened to be negative for the analytes of interest.

3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts

The document describes the reference methods used to establish ground truth for the clinical and archived test sets, rather than explicitly stating "experts" for ground truth.

• For bacterial analytes: Quantitative reference culture (qRefCx) performed at a central reference laboratory. The method was considered positive if the organism was recovered and enumerated at ≥10^3.5 CFU/mL. Molecular methods (single PCR followed by quantitative molecular assay and sequencing (qMol)) were also used in discrepancy resolution.
• For atypical bacteria and viruses: Two conventional PCR assays followed by bidirectional sequencing. A specimen was positive if bidirectional sequencing data matched organism-specific sequences in the NCBI GenBank database.
• For antimicrobial resistance genes: Single PCR assay followed by sequencing for specimens where an applicable bacterium was detected by FilmArray.

The document does not specify the number or qualifications of the individuals performing these reference methods. However, the involvement of a "central reference laboratory" for qRefCx and detailed molecular methods suggests that qualified laboratory personnel with expertise in microbiology and molecular diagnostics would have performed these tasks.

4. Adjudication method for the test set

• Discrepancy Investigation: For clinical study results, discrepancies between FilmArray Pneumonia Panel results and comparator method results were investigated.
  • For bacterial analytes and qRefCx discrepancies: investigated whether FilmArray or qRefCx reported "negative" or "Not Detected" due to being below detection threshold. If unresolved, qMol testing results were considered. If still unresolved, multiple additional molecular assays followed by sequence analysis were used. Standard of Care (SOC) testing results were also considered.
  • For atypical bacteria, viruses, and AMR genes (where molecular comparators were primary): The discrepancy investigation involved further molecular assays (e.g., additional molecular methods, retesting with FilmArray Pneumonia Panel, or sequencing).

The document details the steps taken for resolving discrepancies but doesn't explicitly refer to an "adjudication panel" or specific "X+Y" method with named experts. The process is a multi-step laboratory investigation rather than a formalized expert adjudication panel in the context of imaging.

5. If a multi-reader multi-case (MRMC) comparative effectiveness study was done

No a multi-reader multi-case (MRMC) comparative effectiveness study was mentioned. The study focused on the standalone performance of the device against reference methods, rather than comparing human reader performance with and without AI assistance.

6. If a standalone (i.e., algorithm only without human-in-the-loop performance) was done

Yes, the studies described are standalone performance evaluations. The FilmArray Pneumonia Panel automates the process from nucleic acid extraction to result interpretation. The reported sensitivity, specificity, PPA, and NPA values represent the performance of the algorithm/device itself in detecting and identifying pathogens and resistance genes, without human interpretation influencing the final binary output (Detected/Not Detected) or semi-quantitative binning.

7. The type of ground truth used

The ground truth used in the studies included:

• Quantitative Reference Culture (qRefCx): For bacterial analytes in clinical specimens.
• PCR/Sequencing: For atypical bacteria, viruses, and antimicrobial resistance genes in clinical and archived specimens. This involved conventional PCR followed by bidirectional sequencing, matching results to NCBI GenBank.
• Known Composition of Contrived Specimens: For contrived specimens, the ground truth was the known presence and concentration of spiked organisms and AMR genes.
• Phenotypic AST methods (e.g., ESBL activity testing, carbapenem susceptibility testing, cefoxitin susceptibility testing): Used to assess correlation with AMR gene results on cultured isolates from clinical specimens.

8. The sample size for the training set

The document does not explicitly state a sample size for a "training set." The FilmArray Pneumonia Panel is a molecular diagnostic test that identifies specific nucleic acid sequences. Its development likely involves extensive analytical validation (e.g., inclusivity, exclusivity, LoD, precision), which informs the assay design and algorithm parameters. This process differs from machine learning algorithms that typically require large labeled training sets. The document focuses on performance testing rather than algorithm training.

9. How the ground truth for the training set was established

As no specific "training set" is described in the context of machine learning, the establishment of "ground truth" for training purposes is not directly detailed. However, the analytical studies (e.g., inclusivity, exclusivity, LoD) serve to validate the design and performance characteristics of the assays within the panel. For these analytical studies, ground truth was established by:

• Known concentrations of organisms/nucleic acids: Pure cultures and nucleic acid extracts at defined concentrations.
• Genetically characterized isolates: Using a diverse collection of confirmed strains/isolates/serotypes for inclusivity testing, whose identity and gene presence were validated through standard microbiological and molecular methods (e.g., sequencing, typing).
• In silico analysis: Predicting reactivity based on publicly available genetic sequences.

This rigorous analytical validation process ensures that the fundamental components of the FilmArray Pneumonia Panel (primers, probes, thermal cycling conditions, melting curve analysis parameters) are robust and accurate for their intended targets across known genetic diversity.

Ask a specific question about this device

The Unyvero LRT Application is a qualitative nucleic acid multiplex test intended for the simultaneous detection and identification of nucleic acid sequences from the following microorganisms and antibiotic resistance markers in endotracheal aspirates from adult hospitalized patients with suspected lower respiratory tract infections.

The Unyvero LRT Application performed on the Unyvero System is indicated as an aid in the diagnosis of lower respiratory tract infection in adult hospitalized patients with signs and symptoms of lower respiratory infection: results should be used in conjunction with other clinical and laboratory findings. As tracheal aspirates commonly contain colonizing microorganisms, detection of Unyvero LRT microbial targets does not indicate that the microorganism is the cause of the disease. Unyvero positive results do not rule out co-infection with microorganisms not detected by the Unyvero LRT Application. Negative results do not preclude lower respiratory infection, as the causative agent may be a microorganism not detected by this test.

A negative result for any antibiotic resistance marker does not indicate that detected microorganisms are susceptible to applicable antimicrobial agents. Detected resistance markers cannot be definitively linked to specific microorganisms, and may be present in organisms that are not detected by the Unyvero LRT Application such as organisms present as colonizing or normal flora.

Microbiology cultures of aspirates should be performed to obtain isolates for species identification and antimicrobial susceptibility testing, to differentiate quantities of identified microorganisms as well as normal flora present in the specimen and to identify potential microorganisms not targeted by the Unyvero LRT Application.

The Unyvero LRT Application is a qualitative test that includes specimen processing, genomic bacterial DNA isolation and purification, multiplex PCR and array hybridization and detection. The Unyvero LRT Application performed using the Unyvero System detects specific nucleic acid sequences from microorganisms and resistance markers in tracheal aspirates collected from patients with signs and symptoms of lower respiratory infection.

The Unyvero LRT Application consists of the following components:

• Unyvero LRT Cartridge: Contains DNA isolation and purification reagents, a DNA isolation column, eight separate PCR chambers with eight corresponding detection arrays. The Cartridge also contains fluorescently-labeled primers, hybridization and wash buffers and oligonucleotide probes for detection of targeted PCR products using array hybridization technology.
• Unyvero T1 Sample Tube: Contains glass beads and buffers to lyse bacteria and liquefy the sample.
• Unyvero T1 Sample Tube Cap (with Internal Control): Contains proteinase K and a synthetic internal control gene for process monitoring. The T1 Sample Tube Cap seals the Unyvero Sample Tube after which the internal control is combined with each patient specimen. The internal control DNA sequence does not have significant homology to targeted sequences and is amplified independently in each of the eight PCR chambers and the amplified internal control product is hybridized on each array.
• Unyvero M1 Master Mix: Contains reagents for DNA amplification.
• Unyvero T1 Transfer Tool: The Transfer tool can be used to transfer viscous specimens from the primary sample container to the Unyvero Sample Tube.

The Unyvero System consists of the following components:

• Unyvero Lysator: The Lysator lyses the specimen and can process up to four specimens simultaneously in four separate slots.
• Unyvero Analyzer: The Analyzer automates DNA purification, amplification and detection. Each Analyzer can simultaneously process up to two Unyvero Cartridges with each slot available using random access.
• Unyvero Cockpit: The Cockpit provides the main user interface for the Unyvero System, guides the user through the steps to run the Unvvero LRT Application and automatically generates and displays test results. The Cockpit is equipped with a high-resolution touch screen and a barcode reader.
• Unyvero Sample Tube Holder: The Sample Tube holder holds the Sample Tube securely while the specimen is transferred into the Sample Tube.

Here's a breakdown of the acceptance criteria and study proving the device meets them, based on the provided text:

Device: Unyvero Lower Respiratory Tract (LRT) Application and Unyvero System (Qualitative nucleic acid amplification assay)

Purpose: Simultaneous detection and identification of nucleic acid sequences from specified microorganisms and antibiotic resistance markers in endotracheal aspirates from adult hospitalized patients with suspected lower respiratory tract infections.

1. Table of Acceptance Criteria and Reported Device Performance

The document does not explicitly state pre-defined acceptance criteria in a quantitative table format (e.g., "PPA must be >90%"). However, it describes the performance observed in the validation studies and then concludes that this performance is "acceptable" in conjunction with various limitations and recommended practices. The implicit acceptance criteria appear to be the demonstrated performance characteristics from the clinical studies.

Below is a table summarizing the reported device performance for organism detection (compared to a composite comparator, which is generally the most stringent comparison) and resistance marker detection (with software masking). The document doesn't provide a single, overarching table of "acceptance criteria" but rather details performance metrics across various studies.

Table of Reported Device Performance (Key Clinical Metrics)

2. Sample Size Used for the Test Set and Data Provenance

• Test Set (Clinical Studies):
  • Prospective Study: 603 evaluable tracheal aspirate specimens.
    • Provenance: Multi-center study at nine clinical sites in the United States.
    • Nature: Prospectively collected.
  • Retrospective (Archived) Study: 369 evaluable archived specimens (211 from US study sites, 158 from other US or European sites tested in-house at Curetis).
    • Provenance: US and European sites.
    • Nature: Previously frozen, retrospective.
  • Contrived Clinical Study: Ranged from 21 to 50 specimens for each microorganism or resistance marker.
    • Provenance: Prepared and tested at three sites in the United States and in-house at Curetis.
    • Nature: Contrived specimens (spiked into natural tracheal aspirate matrices).

3. Number of Experts Used to Establish Ground Truth for the Test Set and Qualifications of those Experts

The document does not specify the number or qualifications of experts involved in establishing the ground truth for the clinical studies.

• For "typical" microorganisms in the prospective study, the primary ground truth was standard-of-care (SoC) tracheal aspirate culture. For the composite comparator, it also included independent and validated multiplexed PCR assays with bi-directional sequencing.
• For "atypical" microorganisms, the ground truth was two independent and validated multiplex PCR assays with bi-directional sequencing.
• For antibiotic resistance markers, the ground truth was validated multiplex PCR assays followed by bi-directional sequencing.
• For comparative phenotypic analysis of mecA, phenotypic antimicrobial susceptibility (AST) testing results from established culture methods were used.

The validation of these comparator methods (LoDs, inclusivity) is mentioned, implying expert oversight, but specific expert count or detailed qualifications are not provided.

4. Adjudication Method for the Test Set

The document describes the "Composite Comparator" method as an adjudication of sorts:

• For 'typical' microorganisms: A specimen was considered positive if culture was positive OR if the validated comparator PCR and follow-up bi-directional sequencing was positive. Any specimen negative by both culture and PCR was considered negative. This is a form of "consensus" or "best clinical practice" ground truth, where molecular confirmation is used to bolster culture results, or establish positivity where culture might miss (e.g., non-viable organisms, difficult to culture).
• For 'atypical' microorganisms: Specimens positive by either of the two PCR/sequencing assays were considered positive. Those negative by both were considered negative. This implies a "molecular consensus" approach.
• For 'false positives' against culture: False positive LRT results were analyzed with molecular assays (PCR/bi-directional sequencing) using sample DNA extracts to confirm presence or absence of microorganisms. This acts as a re-adjudication/confirmation step for discordant results.

There is no mention of human expert adjudication panels in the way it might be done for image-based AI devices (e.g., 2+1 radiologist review). The adjudication is inherently built into the definition of the reference/comparator method, often relying on molecular methods to resolve discrepancies or provide more sensitive ground truth than traditional culture.

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was done

No, an MRMC comparative effectiveness study was not done for this device. This device is a diagnostic assay for molecular detection, not an AI-assisted diagnostic aid for human readers/interpreters in the typical MRMC study design (e.g., radiology AI). The study proves the standalone performance of the device.

6. If a Standalone (algorithm only without human-in-the-loop performance) was done

Yes, the study primarily evaluates the standalone performance of the Unyvero LRT Application. The reported PPA and NPA values for specific targets are measures of the device's accuracy in detecting nucleic acid sequences in the specimens, independent of human interpretation of the device's raw output. The device itself (Unyvero System with LRT Application) automates DNA purification, amplification, and detection, and automatically generates and displays results. The human involvement is in sample preparation and loading, and interpreting the final qualitative "positive/negative" result the device provides, rather than interpreting raw data alongside or without AI assistance.

7. The Type of Ground Truth Used

The ground truth used varied depending on the target:

• For 'typical' microorganisms:
  • Primary Reference: Standard-of-care (SoC) tracheal aspirate culture.
  • Composite Comparator: A combination of SoC culture PLUS independent and validated multiplexed PCR assays with bi-directional sequencing. This represents a more sensitive and comprehensive "true presence" ground truth, especially for organisms that may be difficult to culture or present as non-viable DNA.
• For 'atypical' microorganisms: Two independent and validated multiplex PCR assays with bi-directional sequencing.
• For antibiotic resistance markers: Validated multiplex PCR assays followed by bi-directional sequencing.
• For genotypic linkage and phenotypic correlation: Molecular sequencing of cultured isolates for the presence of the resistance marker gene, and phenotypic antimicrobial susceptibility testing (AST) results from those isolates.

8. The Sample Size for the Training Set

The document does not explicitly mention a "training set" in the context of machine learning or AI models. Given that this is a PCR-based diagnostic assay, it's developed through traditional molecular assay design and optimization, not typically through supervised machine learning with distinct training and test sets as seen in AI imaging. The "development" or "optimization" of the assay (e.g., primer design, LoD determination, inclusivity/exclusivity testing) would constitute its "training" or optimization phase, but no sample size for this phase is provided. The data presented relates to the validation of the final assay.

9. How the Ground Truth for the Training Set Was Established

As noted above, there's no explicitly defined "training set" in the machine learning sense. The ground truth for the analytical studies and clinical validation studies (which would inform refinement if developmental iterations were considered "training") was established as follows:

• Analytical Studies (LoD, Reproducibility, Inclusivity, Exclusivity, Interference): Samples were contrived by spiking known concentrations of well-characterized reference strains (ATCC, NCTC, DSM, JMI, Micromyx, NRZ, UCLA, RKI, clinical isolates) into artificial respiratory matrix (ARM) or PBS. The "ground truth" here is the known identity and concentration of the spiked microorganisms/resistance markers.
• Clinical Studies (Prospective and Retrospective): As detailed in point 7, ground truth was established by:
  • Standard-of-care (SoC) tracheal aspirate culture.
  • Validated multiplex PCR assays with bi-directional sequencing.
  • A composite of SoC culture and PCR/sequencing.
  • Molecular sequencing of isolates and phenotypic AST.

This means the "ground truth" was established by conventional microbiological and molecular diagnostic methods, which were considered the gold standard for comparing the performance of the new Unyvero system.

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