FilmArray Pneumonia Panel

{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 {4}------------------------------------------------ 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. {5}------------------------------------------------ This section applies only to requirements of the Paperwork Reduction Act of 1995. ## *DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.* The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to: > Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff@fda.hhs.gov "An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number." {6}------------------------------------------------ # 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<br>complex | Klebsiella oxytoca | Serratia marcescens | | Enterobacter cloacae complex | Klebsiella pneumoniae group | Staphylococcus aureus | | Haemophilus influenzae | Moraxella catarrhalis | Streptococcus agalactiae | | Escherichia coli | 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 | Respiratory Syncytial Virus | | Human Metapneumovirus | Influenza B | | | Antimicrobial Resistance Genes | | | | CTX-M | NDM | <i>mecA/C</i> and MREJ | | IMP | OXA-48-like | | | KPC | VIM | | 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 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 | | Streptococcus agalactiae | Human Rhinovirus/Enterovirus | | Streptococcus pneumoniae | Influenza A | | Streptococcus pyogenes | Influenza B | | 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 {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:<br>FilmArray Pneumonia Panel | Predicate:<br>Curetis Unyvero LRT Application<br>(DEN170047) | |-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------| | Specimen<br>Types | Sputum-like (induced or expectorated<br>sputum, endotracheal aspirates) and BAL-<br>like (BAL or mini-BAL) specimens from<br>individuals (of all ages) with signs and/or<br>symptoms of lower respiratory tract<br>infection | Endotracheal aspirate specimens from adult<br>hospitalized patients with suspected lower<br>respiratory tract infections | | Element | New Device:<br>FilmArray Pneumonia Panel | Predicate:<br>Curetis Unyvero LRT Application<br>(DEN170047) | | Organisms<br>Detected | Bacteria: Acinetobacter calcoaceticus-<br>baumannii complex, Enterobacter cloacae<br>complex, Escherichia coli , Haemophilus<br>influenzae, Klebsiella aerogenes, Klebsiella<br>oxytoca, Klebsiella pneumoniae group,<br>Moraxella catarrhalis, Proteus spp.,<br>Pseudomonas aeruginosa, Serratia<br>marcescens, Staphylococcus aureus,<br>Streptococcus agalactiae, Streptococcus<br>pneumoniae, Streptococcus pyogenes<br>Atypical Bacteria: Chlamydia pneumoniae,<br>Legionella pneumophila, Mycoplasma<br>pneumoniae<br>Antimicrobial Resistance Genes: CTX-M,<br>IMP, KPC, $mecA/C$ + MREJ, NDM,<br>OXA48-like, VIM | Bacteria: Acinetobacter spp., Citrobacter<br>freundii, Enterobacter cloacae complex,<br>Escherichia coli, Haemophilus influenzae,<br>Klebsiella oxytoca, Klebsiella pneumoniae,<br>Klebsiella variicola, Moraxella catarrhalis,<br>Morganella morganii, Proteus spp.,<br>Pseudomonas aeruginosa, Serratia<br>marcescens, Staphylococcus aureus,<br>Stenotrophomonas maltophilia,<br>Streptococcus pneumoniae<br>Atypical Bacteria: Chlamydia pneumoniae,<br>Legionella pneumophila, Mycoplasma<br>pneumonia,<br>Antimicrobial Resistance Genes: CTX-M,<br>KPC, mecA, NDM, OXA-23, OXA-24,<br>OXA-48, OXA-58, TEM, VIM | | Viruses: Adenovirus, Coronavirus, Human<br>Metapneumovirus, Human<br>Rhinovirus/Enterovirus, Influenza A,<br>Influenza B, Parainfluenza virus,<br>Respiratory Syncytial virus | | | | Analyte | DNA/RNA | DNA only | | Technological<br>Principles | Multiplex nucleic acid | Same | | Result types | Semi-quantitative detection with bin values<br>(in 1-log rounded copies/mL bins) for<br>Bacteria<br>Qualitative for Atypical Bacteria,<br>Antimicrobial Resistance Genes, and<br>Viruses | Qualitative for all analytes | | Instrumentation | FilmArray, FilmArray 2.0, or FilmArray<br>Torch | Unyvero System | | Time to result | About 1 hour | About 4-5 hours | | Reagent<br>Storage | Room temperature | Unknown | | Test<br>Interpretation | Automated test interpretation and report<br>generation. User cannot access raw data. | Same | | Controls | Two controls are included in each reagent<br>pouch to control for sample processing and<br>both stages of PCR and melt analysis. | One control is included in each cartridge to<br>monitor for the presence of PCR inhibitors<br>and enables the system to detect any failures<br>in the testing process. | | User<br>Complexity | Moderate/Low | Same | Table 1. Comparison of the FilmArray Pneumonia Panel and the Curetis Unyvero LRT Application) {10}------------------------------------------------ {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 | | | | | | | | | | | | |-----------------------------------------------------------------------------------------------------|---------------|-----------|----------|---------|-----------|----------|----------|----------|----------|----------|--| | | | Overall | Site 1ª | Site 2 | Site 3 | Site 4 | Site 5 | Site 6 | Site 7 | Site 8 | | | Sex | Male | 480 (57%) | 80 (59%) | 7 (54%) | 138 (55%) | 21 (68%) | 75 (61%) | 82 (52%) | 27 (61%) | 50 (55%) | | | | Female | 366 (43%) | 55 (41%) | 6 (46%) | 113 (45%) | 10 (32%) | 48 (39%) | 76 (48%) | 17 (39%) | 41 (45%) | | | Age | ≤ 5 years | 23 (3%) | 0 (0%) | 5 (38%) | 0 (0%) | 15 (48%) | 0 (0%) | 3 (2%) | 0 (0%) | 0 (0%) | | | | 6 - 17 years | 27 (3%) | 0 (0%) | 8 (62%) | 0 (0%) | 13 (42%) | 0 (0%) | 4 (3%) | 1 (2%) | 1 (1%) | | | | 18 - 34 years | 70 (8%) | 18 (13%) | 0 (0%) | 17 (7%) | 3 (10%) | 10 (8%) | 10 (6%) | 5 (11%) | 7 (8%) | | | | 35 - 65 years | 470 (56%) | 78 (58%) | 0 (0%) | 152 (61%) | 0 (0%) | 70 (57%) | 88 (56%) | 27 (61%) | 55 (60%) | | | | > 65 years | 255 (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%) | | | | | | | | | | | | | Outpatient | | 159 (19%) | 18 (13%) | 0 (0%) | 28 (11%) | 22 (71%) | 31 (25%) | 39 (25%) | 14 (32%) | 7 (8%) | | | Emergency | | 21 (2%) | 1 (1%) | 1 (8%) | 0 (0%) | 0 (0%) | 10 (8%) | 1 (1%) | 5 (11%) | 3 (3%) | | | Total | | 846 | 135 | 13 | 251 | 31 | 123 | 158 | 44 | 91 | | 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 | | | | | | | | | | | |--------------|---------------|-----------|-----------|----------|-----------|-----------|----------|----------|----------|-----------| | | | Overall | Site 1 | Site 2 | Site 3 | Site 4 | Site 5 | Site 6 | Site 7 | Site 8 | | Sex | Male | 481 (58%) | 66 (59%) | 54 (54%) | 136 (56%) | 97 (61%) | 14 (82%) | 31 (53%) | 34 (74%) | 49 (47%) | | | Female | 355 (42%) | 45 (41%) | 46 (46%) | 105 (44%) | 61 (39%) | 3 (18%) | 28 (47%) | 12 (26%) | 55 (53%) | | Age | ≤ 5 years | 138 (17%) | 0 (0%) | 49 (49%) | 0 (0%) | 80 (51%) | 0 (0%) | 0 (0%) | 2 (4%) | 7 (7%) | | | 6 - 17 years | 107 (13%) | 0 (0%) | 35 (35%) | 0 (0%) | 64 (41%) | 0 (0%) | 0 (0%) | 2 (4%) | 6 (6%) | | | 18 - 34 years | 86 (10%) | 15 (14%) | 16 (16%) | 20 (8%) | 13 (8%) | 1 (6%) | 6 (10%) | 5 (11%) | 10 (10%) | | | 35 - 65 years | 284 (34%) | 51 (46%) | 0 (0%) | 133 (55%) | 1 (1%) | 6 (35%) | 36 (61%) | 20 (43%) | 37 (36%) | | | > 65 years | 221 (26%) | 45 (41%) | 0 (0%) | 88 (37%) | 0 (0%) | 10 (59%) | 17 (29%) | 17 (37%) | 44 (42%) | | Care Setting | Hospitalized | 682 (82%) | 106 (95%) | 64 (64%) | 219 (91%) | 105 (66%) | 12 (71%) | 52 (88%) | 23 (50%) | 101 (97%) | | | Outpatient | 73 (9%) | 2 (2%) | 14 (14%) | 18 (7%) | 24 (15%) | 2 (12%) | 5 (8%) | 7 (15%) | 1 (1%) | | | Emergency | 81 (10%) | 3 (3%) | 22 (22%) | 4 (2%) | 29 (18%) | 3 (18%) | 2 (3%) | 16 (35%) | 2 (2%) | {12}------------------------------------------------ | Sputum | | | | | | | | | | |--------|---------|--------|--------|--------|--------|--------|--------|--------|--------| | | Overall | Site 1 | Site 2 | Site 3 | Site 4 | Site 5 | Site 6 | Site 7 | Site 8 | | Total | 836 | 111 | 100 | 241 | 158 | 17 | 59 | 46 | 104 | 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 | | | | |---------|-----------|-----------------|-----------------| | Analyte | Reference | Sensitivity/PPA | Specificity/NPA | {13}------------------------------------------------ | | Method | TP/(TP + FN) | % | 95%CI | TN/(TN + FP) | % | 95%CI | |------------------------------------------------|---------|-------------------|------|------------|--------------|------|------------| | | | Bacteria | | | | | | | Acinetobacter calcoaceticus-baumannii complexb | qRefCx | 0/0 | - | | 839/846 | 99.2 | 98.3-99.6% | | Klebsiella aerogenesc | qRefCx | 6/7 | 85.7 | 48.7-97.4% | 832/839 | 99.2 | 98.3-99.6% | | Enterobacter cloacae complexd | qRefCx | 11/12 | 91.7 | 64.6-98.5% | 822/834 | 98.6 | 97.5-99.2% | | Escherichia colie | qRefCx | 12/12 | 100 | 75.8-100% | 826/834 | 99.0 | 98.1-99.5% | | Haemophilus influenzaef | qRefCx | 10/10 | 100 | 72.2-100% | 764/836 | 91.4 | 89.3-93.1% | | Klebsiella oxytocag | qRefCx | 2/2 | 100 | 34.2-100% | 835/844 | 98.9 | 98.0-99.4% | | Klebsiella pneumoniae grouph | qRefCx | 15/15 | 100 | 79.6-100% | 819/831 | 98.6 | 97.5-99.2% | | Moraxella catarrhalisi | qRefCx | 0/0 | - | | 817/846 | 96.6 | 95.1-97.6% | | Proteus spp.j | qRefCx | 5/5 | 100 | 56.6-100% | 837/841 | 99.5 | 98.8-99.8% | | Pseudomonas aeruginosak | qRefCx | 36/36 | 100 | 90.4-100% | 772/810 | 95.3 | 93.6-96.6% | | Serratia marcescensl | qRefCx | 6/6 | 100 | 61.0-100% | 834/840 | 99.3 | 98.5-99.7% | | Staphylococcus aureusm | qRefCx | 46/47 | 97.9 | 88.9-99.6% | 729/799 | 91.2 | 89.1-93.0% | | Streptococcus agalactiaen | qRefCx | 1/1 | 100 | - | 821/845 | 97.2 | 95.8-98.1% | | Streptococcus pneumoniaeo | qRefCx | 5/5 | 100 | 56.6-100% | 817/841 | 97.1 | 95.8-98.1% | | Streptococcus pyogenesp | qRefCx | 2/2 | 100 | 34.2-100% | 838/844 | 99.3 | 98.5-99.7% | | | | Atypical Bacteria | | | | | | | Chlamydia pneumoniaeq | PCR/Seq | 0/0 | - | - | 844/845 | 99.9 | 99.3-100% | | Legionella pneumophila | PCR/Seq | 2/2 | 100 | 34.2-100% | 833/833 | 100 | 99.5-100% | | Mycoplasma pneumoniaer | PCR/Seq | 3/3 | 100 | 43.9-100% | 841/842 | 99.9 | 99.3-100% | | | | Viruses | | | | | | | Adenovirus | PCR/Seq | 8/8 | 100 | 67.6-100% | 837/837 | 100 | 99.5-100% | | Coronaviruss | PCR/Seq | 18/21 | 85.7 | 65.4-95.0% | 810/823 | 98.4 | 97.3-99.1% | | Human Metapneumovirust | PCR/Seq | 8/8 | 100 | 67.6-100% | 836/837 | 99.9 | 99.3-100% | | Human Rhinovirus/Enterovirusu | PCR/Seq | 52/54 | 96.3 | 87.5-99.0% | 771/782 | 98.6 | 97.5-99.2% | | Influenza Av | PCR/Seq | 10/10 | 100 | 72.2-100% | 830/833 | 99.6 | 98.9-99.9% | | Influenza Bw | PCR/Seq | 5/6 | 83.3 | 43.6-97.0% | 837/838 | 99.9 | 99.3-100% | | Parainfluenza Virusx | PCR/Seq | 16/18 | 88.9 | 67.2-96.9% | 824/826 | 99.8 | 99.1-99.9% | | Respiratory Syncytial Virus | PCR/Seq | 3/3 | 100 | 43.9-100% | 841/841 | 100 | 99.5-100% | " 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 | | | | | | | | |------------------------------------------------|---------------------|-----------------|------|------------|-----------------|------|------------| | Analyte | Reference<br>Method | TP/(TP +<br>FN) | % | 95%CI | TN/(TN +<br>FP) | % | 95%CI | | <b>Bacteria</b> | | | | | | | | | Acinetobacter calcoaceticus-baumannii complexb | qRefCx | 10/11 | 90.9 | 62.3-98.4% | 807/825 | 97.8 | 96.6-98.6% | | Klebsiella aerogenesc | qRefCx | 3/4 | 75.0 | 30.1-95.4% | 823/832 | 98.9 | 98.0-99.4% | | Enterobacter cloacae complexd | qRefCx | 11/12 | 91.7 | 64.6-98.5% | 803/824 | 97.5 | 96.1-98.3% | | Escherichia colie | qRefCx | 23/24 | 95.8 | 79.8-99.3% | 787/812 | 96.9 | 95.5-97.9% | | Haemophilus influenzaef | qRefCx | 16/18 | 88.9 | 67.2-96.9% | 727/818 | 88.9 | 86.5-90.9% | | Klebsiella oxytocag | qRefCx | 9/9 | 100 | 70.1-100% | 817/827 | 98.8 | 97.8-99.3% | | Klebsiella pneumoniae grouph | qRefCx | 21/23 | 91.3 | 73.2-97.6% | 769/813 | 94.6 | 92.8-95.9% | | Moraxella catarrhalisi | qRefCx | 5/5 | 100 | 56.6-100% | 761/831 | 91.6 | 89.5-93.3% | | Proteus spp.j | qRefCx | 15/15 | 100 | 79.6-100% | 813/821 | 99.0 | 98.1-99.5% | | Pseudomonas aeruginosak | qRefCx | 103/106 | 97.2 | 92.0-99.0% | 673/730 | 92.2 | 90.0-93.9% | | Serratia marcescensl | qRefCx | 26/27 | 96.3 | 81.7-99.3% | 782/809 | 96.7 | 95.2-97.7% | | Staphylococcus aureusm | qRefCx | 111/112 | 99.1 | 95.1-99.8% | 631/724 | 87.2 | 84.5-89.4% | | Streptococcus agalactiaen | qRefCx | 9/9 | 100 | 70.1-100% | 793/827 | 95.9 | 94.3-97.0% | | Streptococcus pneumoniaeo | qRefCx | 16/16 | 100 | 80.6-100% | 785/820 | 95.7 | 94.1-96.9% | | Streptococcus pyogenesp | qRefCx | 6/6 | 100 | 61.0-100% | 825/830 | 99.4 | 98.6-99.7% | | <b>Atypical Bacteria</b> | | | | | | | | | Chlamydia pneumoniae | PCR/Seq | 0/0 | - | - | 835/835 | 100 | 99.5-100% | | Legionella pneumophilaq | PCR/Seq | 0/1 | 0 | - | 826/826 | 100 | 99.5-100% | | Mycoplasma pneumoniaer | PCR/Seq | 7/8 | 87.5 | 52.9-97.8% | 827/827 | 100 | 99.5-100% | | <b>Viruses</b> | | | | | | | | | Adenoviruss | PCR/Seq | 13/17 | 76.5 | 52.7-90.4% | 815/817 | 99.8 | 99.1-99.9% | | Coronavirust | PCR/Seq | 28/32 | 87.5 | 71.9-95.0% | 796/802 | 99.3 | 98.4-99.7% | | Human Metapneumovirusu | PCR/Seq | 20/21 | 95.2 | 77.3-99.2% | 812/813 | 99.9 | 99.3-100% | | Human Rhinovirus/Enterovirusv | PCR/Seq | 96/96 | 100 | 96.2-100% | 717/730 | 98.2 | 97.0-99.0% | | Influenza Aw | PCR/Seq | 13/13 | 100 | 77.2-100% | 819/822 | 99.6 | 98.9-99.9% | | Influenza Bx | PCR/Seq | 12/12 | 100 | 75.8-100% | 821/823 | 99.8 | 99.1-99.9% | {15}------------------------------------------------ | Sputum | | | | | | | | |------------------------------|---------------------|---------------------------------|------|------------|---------------------------------|------|------------| | Analyte | Reference<br>Method | Sensitivity/PPA<br>TP/(TP + FN) | % | 95%CI | Specificity/NPA<br>TN/(TN + FP) | % | 95%CI | | Parainfluenza Virusy | PCR/Seq | 28/29 | 96.6 | 82.8-99.4% | 804/806 | 99.8 | 99.1-99.9% | | Respiratory Syncytial Virusz | PCR/Seq | 43/43 | 100 | 91.8-100% | 787/791 | 99.5 | 98.7-99.8% | " 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). | | BAL | | | Sputum | | | | | | | | | |-------------------------------|---------------------|------|----------------|---------------------|------|----------------|---------------------|------|----------------|---------------------|------|----------------| | | PPA | | NPA | PPA | | NPA | | | | | | | | Analyte | TP/<br>(TP +<br>FN) | % | 95%CI | TN/<br>(TN +<br>FP) | % | 95%CI | TP/<br>(TP +<br>FN) | % | 95%CI | TN/<br>(TN +<br>FP) | % | 95%CI | | CTX-<br>Mb | 6/7 | 85.7 | 48.7-<br>97.4% | 144/144 | 100 | 97.4-<br>100% | 8/10 | 80.0 | 49.0-<br>94.3% | 280/281 | 99.6 | 98.0-<br>99.9% | | IMP | 0/0 | - | - | 151/151 | 100 | 97.5-<br>100% | 0/0 | - | - | 291/291 | 100 | 98.7-<br>100% | | KPCc | 2/2 | 100 | 34.2-<br>100% | 148/149 | 99.3 | 96.3-<br>99.9% | 7/7 | 100 | 64.6-<br>100% | 284/284 | 100 | 98.7-<br>100% | | <i>mecA/C</i><br>and<br>MREJd | 40/45 | 88.9 | 76.5-<br>95.2% | 64/70 | 91.4 | 82.5-<br>96.0% | 94/98 | 95.9 | 90.0-<br>98.4% | 91/104 | 87.5 | 79.8-<br>92.5% | | NDMe | 0/1 | 0 | - | 149/150 | 99.3 | 96.3-<br>99.9% | 0/0 | - | - | 291/291 | 100 | 98.7-<br>100% | | OXA-<br>48-<br>like | 0/0 | - | - | 151/151 | 100 | 97.5-<br>100% | 0/0 | - | - | 291/291 | 100 | 98.7-<br>100% | | VIMf | 0/0…