Who is the manufacturer of FilmArray Pneumonia Panel?

Biofire Diagnostics, LLC filed the 510(k) submission for FilmArray Pneumonia Panel (K180966).

What is the FDA product code for FilmArray Pneumonia Panel?

QDP. It is classified under 21 CFR 866.3985 as a Class 2 device in the Microbiology panel.

What is the regulatory pathway for FilmArray Pneumonia Panel?

510(k) clearance (submission type: Traditional).

Who can help prepare an FDA 510(k) submission like FilmArray Pneumonia Panel?

Innolitics — a medical-device software consultancy — provides regulatory and engineering support for FDA 510(k) submissions. See https://innolitics.com/services/510ks/ or contact https://innolitics.com/contact.

FilmArray Pneumonia Panel

Q: What are the predicate devices for FilmArray Pneumonia Panel?

Curetis Unyvero LRT Application (DEN170047).

K180966 · Biofire Diagnostics, LLC · QDP · Nov 9, 2018 · Microbiology

Device Facts

Record ID	K180966
Device Name	FilmArray Pneumonia Panel
Applicant	Biofire Diagnostics, LLC
Product Code	QDP · Microbiology
Decision Date	Nov 9, 2018
Decision	SESE
Submission Type	Traditional
Regulation	21 CFR 866.3985
Device Class	Class 2
Attributes	Real-World Evidence, Pediatric

Real-World Evidence

Submission	Device	Sponsor	RWD Sources	RWE Use Summary	Key Tags
K180966 · Nov 9, 2018	FilmArray Pneumonia Panel	Biofire Diagnostics, LLC	Residual clinical specimens (BAL and sputum) from patients suspected of lower respiratory tract infection	Residual clinical specimens were used to establish the clinical performance of the FilmArray Pneumonia Panel in a multi-center study.	Residual clinical specimens; Clinical performance study; Lower respiratory tract infection

Clinical Evidence

Study Design	Population	Comparator	Key Endpoints
Prospective clinical study; Multi-center prospective clinical study; Follow-up/Duration: October 2016 to July 2017; Study Period: October 2016 to July 2017	Individuals suspected of lower respiratory tract infection; Sample Size: 846 BAL and 836 sputum specimens; Number of Sites: 8	Quantitative reference culture (qRefCx) and PCR/sequencing-based comparator methods	Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA)

AI Performance

Output	Algorithm	Acceptance	Observed	Dev DS	Dev Readers	Test DS	Test Readers
Detection of respiratory viral and bacterial nucleic acids and antimicrobial resistance genes	Nested multiplex PCR with melt curve analysis	PPA ≥ 90% (80% lower bound 95% CI) and NPA ≥ 98% (95% lower bound 95% CI)	PPA and NPA varied by analyte; see clinical performance tables for specific values	—	—	Prospective clinical study: 846 BAL and 836 sputum specimens; Archived retrospective specimens: 171 specimens; Contrived specimens: 1125 specimens	—

Indications for Use

The FilmArray® Pneumonia Panel is a multiplexed nucleic acid test intended for use with FilmArray® 2.0, or FilmArray® Torch systems for the simultaneous detection of multiple respiratory viral and bacterial nucleic acids, as well as select antimicrobial resistance genes, in sputum-like specimens (induced or expectorated sputum, or endotracheal aspirates) or bronchoalveolar lavage (BAL)-like specimens (BAL or mini-BAL) obtained from individuals suspected of lower respiratory tract infection. The following bacteria are reported semi-quantitatively with bins representing approximately 10^4, 10^5, 10^6, or ≥10^7 genomic copies of bacterial nucleic acid per milliliter (copies/mL) of specimen, to aid in estimating relative abundance of nucleic acid from these common bacteria within a specimen: Bacteria reported with bins of 10^4, 10^5, 10^6, or ≥10^7 copies/mL: Acinetobacter calcoaceticus-baumannii complex, Enterobacter cloacae complex, Escherichia coli, Haemophilus influenzae, Klebsiella aerogenes, Klebsiella oxytoca, Klebsiella pneumoniae group, Moraxella catarrhalis, Proteus spp., Pseudomonas aeruginosa, Serratia marcescens, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus pneumoniae, Streptococcus pyogenes. The following atypical bacteria, viruses, and antimicrobial resistance genes are reported qualitatively: Atypical Bacteria: Chlamydia pneumoniae, Legionella pneumophila, Mycoplasma pneumoniae. Viruses: Adenovirus, Coronavirus, Human Metapneumovirus, Human Rhinovirus/Enterovirus, Influenza A, Influenza B, Parainfluenza Virus, Respiratory Syncytial Virus. Antimicrobial Resistance Genes: CTX-M, IMP, 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 if used in conjunction with other clinical and epidemiological information. The results of this test should not be used as for diagnosis, treatment, or other patient management decisions. Negative results in the setting of a respiratory illness may be due to infection with pathogens that are not detected by this test, pathogens below the limit of detection, or in the case of bacterial analytes, present at levels below the lowest reported 10^4 copies/mL bin. Detection of analytes does not rule out co-infection with other organisms: the agent(s) detected by the Film Array Pneumonia Panel may not be the definite cause of disease. Additional laboratory testing (e.g. bacterial and viral culture, immunofluorescence, and radiography) may be necessary when evaluating a patient with possible lower respiratory tract infection. Detection of bacterial nucleic acid may be indicative of colonizing or normal respiratory flora and may not indicate the causative agent of pneumonia. Semi-quantitative bin (copies/mL) results generated by the FilmArray Pneumonia Panel are not equivalent to CFU/mL and do not consistently correlate with the quantity of bacterial analytes compared to CFU/mL. For specimens with multiple bacteria detected, the relative abundance of nucleic acids (copies/mL) may not correlate with the relative abundance of bacteria as determined by culture (CFU/mL). Clinical correlation is advised to determine significance of semi-quantitative bin (copies/mL) for clinical management. The antimicrobial resistance gene detected may or may not be associated with the agent(s) responsible for disease. Negative results for these antimicrobial resistance gene assays do not indicate susceptibility to corresponding classes of antimicrobials, as multiple mechanisms of antimicrobial resistance exist. Antimicrobial resistance can occur via multiple mechanisms. A "Not Detected" result for a genetic marker of antimicrobial resistance does not indicate susceptibility to associated antimicrobial drugs or drug classes. A "Detected" result for a genetic marker of antimicrobial resistance cannot be definitively linked to the microorganism(s) detected. Culture is required to obtain isolates for antimicrobial susceptibility testing, and FilmArray Pneumonia Panel results should be used in conjunction with culture results for determination of bacterial susceptibility or resistance. Due to the genetic similarity between human rhinovirus and enterovirus, the test cannot reliably differentiate them. A positive Rhinovirus/Enterovirus result should be followed up using an alternate method (e.g., cell culture or sequence analysis) if differentiation is required. Culture is required to identify pathogens not detected by the FilmArray Pneumonia Panel, to further speciate analytes in genus, complex, or group results if desired, to identify bacterial pathogens present below the 10^4 copies/mL bin if desired, and for antimicrobial susceptibility testing.

Device Story

The FilmArray Pneumonia Panel is a closed-system, multiplexed nucleic acid test for use on FilmArray, FilmArray 2.0, or FilmArray Torch systems. It processes sputum-like or BAL-like specimens to identify 26 pathogens and associated AMR genes. The device uses mechanical/chemical lysis and magnetic bead purification, followed by nested multiplex PCR (first stage: large volume rt-PCR; second stage: singleplex PCR in array wells). Results are generated via melt curve analysis and real-time amplification data, providing qualitative results for viruses/atypical bacteria and semi-quantitative bins (10^4 to ≥10^7 copies/mL) for common bacteria. Used in clinical laboratories by technicians; results aid healthcare providers in diagnosing LRTI and managing antimicrobial therapy in conjunction with culture and clinical data. The device benefits patients by providing rapid (approx. 1 hour) identification of potential pathogens and resistance markers, supporting timely clinical decision-making.

Clinical Evidence

Multi-center prospective study (8 U.S. sites, 2016-2017) evaluated 846 BAL and 836 sputum specimens. Comparator: quantitative reference culture (qRefCx) for bacteria (threshold 10^3.5 CFU/mL) and PCR/sequencing for atypical bacteria/viruses/AMR genes. Sensitivity/PPA and Specificity/NPA reported for all analytes. Additional testing included archived specimens and contrived samples to address low-prevalence analytes. Performance goals generally met; discrepancies investigated via additional molecular methods.

Technological Characteristics

Multiplex nucleic acid test; nested PCR; melt curve analysis. Pouch-based system with integrated lysis, extraction, and amplification. Semi-quantitative binning (10^4-10^7 copies/mL). Compatible with FilmArray 2.0 and Torch systems. Room temperature reagent storage. Software-automated interpretation.

Indications for Use

Indicated for individuals suspected of lower respiratory tract infection (LRTI) exhibiting signs/symptoms of respiratory infection. Used for simultaneous detection/identification of respiratory viral/bacterial nucleic acids and select antimicrobial resistance genes in sputum-like or BAL-like specimens.

Regulatory Classification

Identification

A device to detect and identify microorganisms and associated resistance marker nucleic acids directly from respiratory specimens is an in vitro diagnostic device intended for the detection and identification of microorganisms and associated resistance markers in respiratory specimens collected from patients with signs or symptoms of respiratory infection. The device is intended to aid in the diagnosis of respiratory infection in conjunction with clinical signs and symptoms and other laboratory findings. These devices do not provide confirmation of antibiotic susceptibility since mechanisms of resistance may exist other than those detected by the device.

Special Controls

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

Predicate Devices

Curetis Unyvero LRT Application (DEN170047)

Submission Summary (Full Text)

{0} # 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY A. 510(k) Number: K180966 B. Purpose for Submission: To obtain a substantial equivalence determination for the FilmArray Pneumonia Panel C. Measurand: 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, Chlamydia pneumoniae, Legionella pneumophila, Mycoplasma pneumoniae, Adenovirus, Coronavirus, Human Metapneumovirus, Human Rhinovirus/Enterovirus, Influenza A, Influenza B, Parainfluenza Virus, Respiratory Syncytial Virus, CTX-M, IMP, KPC, NDM, OXA-48-like, VIM, mecA/C and MREJ. D. Type of Test: Qualitative and quantitative nucleic acid amplification assay E. Applicant: BioFire Diagnostics, LLC F. Proprietary and Established Names: FilmArray Pneumonia Panel G. Regulatory Information: 1. Regulation section: 21 CFR 866.3965 - Device to detect and identify microorganisms and associated resistance marker nucleic acids directly in respiratory specimens 2. Classification: Class II (Special Controls) {1} 3. Product code: QBH 4. Panel: 83-Microbiology ### H. Indications for use: 1. Indications for use(s): 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 $ \geq10^{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 $ \geq 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 | Respiratory Syncytial Virus | | Human Metapneumovirus | Influenza B | | | Antimicrobial Resistance Genes | | | | CTX-M | NDM | mecA/C and MREJ | | IMP | OXA-48-like | | | KPC | VIM | | 2 {2} 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 10⁴ 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 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 10⁴ copies/mL bin if desired, and for antimicrobial susceptibility testing. 3 {3} ## 2. Indication(s) for use: Same as Intended Use ## 3. Special conditions for use statement(s): - For prescription use only - For in vitro diagnostic use only ### Limitations: - For prescription use only. - The FilmArray Pneumonia Panel has not been validated for testing of specimens other than unprocessed sputum-like and BAL-like specimens. - Contact or treatment of specimens with decontaminating agents (bleach, MycoPrep (NaOH and NALC), 2% NaOH and 5% Oxalic acid) can cause false negative results (see Interference section). - The performance of the FilmArray Pneumonia Panel has not been established for monitoring treatment of infection. - The effect of antibiotic treatment on test performance including semi-quantitative bin results has not been specifically evaluated. - Viral and bacterial nucleic acids may persist in vivo independent of organism viability. Detection of organism target(s) does not imply that the corresponding organisms are infectious or are the causative agents for clinical symptoms. - The FilmArray Pneumonia Panel results for bacteria are provided as a qualitative Detected/Not Detected result with an associated semi-quantitative bin result of $ 10^{4} $ , $ 10^{5} $ , $ 10^{6} $ , or $ \geq10^{7} $ copies of genomic nucleic acid per milliliter of specimen. An exact quantitative value is not provided. The semi-quantitative (copies/mL) bin result does not distinguish between nucleic acid from live or dead bacteria. - A negative FilmArray Pneumonia Panel result does not exclude the possibility of viral or bacterial infection. Negative test results may occur from the presence of sequence variants in the region targeted by the assay, the presence of inhibitors, technical error, sample mix-up or an infection caused by an organism not detected by the panel. Test results may also be affected by concurrent antiviral/antibacterial therapy or levels of organism in the specimen that are below the limit of detection for the test or below the reportable level for bacterial analytes. Negative results should not be used as the sole basis for diagnosis, treatment, or other patient management decisions. - Concomitant culture of specimens is required with the FilmArray Pneumonia Panel. Culture is needed for recovery of isolates and antimicrobial susceptibility testing, as well as further speciation of genus, complex, or group level results (if desired). - Due to the genetic similarity between human rhinovirus and enterovirus, the FilmArray Pneumonia Panel cannot reliably differentiate them. A FilmArray Pneumonia Panel Human Rhinovirus/Enterovirus Detected 4 {4} result should be followed-up using an alternate method (e.g. cell culture or sequence analysis) if differentiation between the viruses is required. - The in-silico analyses performed to predict amplification and detection of organisms and antimicrobial resistance genes were based on a comparison of target gene sequences available in GenBank to FilmArray Pneumonia Panel primer sequences. In-silico analyses were performed between January 2016 and January 2018. Entries of new sequences added to the database after these dates have not been evaluated. Additional limitations on reactivity may be identified as new sequence data are deposited and/or as new sequence variants emerge. - Based on in-silico analysis, the MREJ assay (which is only reported if Staphylococcus aureus is detected and the mecA/C assay is also positive) is predicted to have impaired reactivity or to be non-reactive with MREJ types ix, xv and xviii, as well as types xix and xx (associated with methicillin-sensitive S. aureus; MSSA), and MREJ sequences annotated from non-aureus Staphylococcus species and non-Staphylococci such as Bacillus cereus, Bacillus thuringiensis, Macrococcus caseolyticus, Clostridium acidurici, and Rummeliibacillus stabekisii. - Positive and negative predictive values are highly dependent on prevalence. False negative test results are more likely during peak activity when prevalence of disease is high. False positive test results are more likely during periods when prevalence is moderate to low. - Performance characteristics for influenza A were established during the 2016-2017 influenza season. When other novel influenza A viruses are emerging, performance characteristics may vary. If infection with a novel influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent influenza viruses and sent to state or local health departments for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens. - Due to the small number of positive specimens collected for certain organisms during the prospective clinical study, performance characteristics for several analytes in one or both matrices were primarily established using archived and/or contrived specimens as detailed in the Clinical Performance section. # 4. Special instrument requirements: The FilmArray Pneumonia Panel is performed on FilmArray, FilmArray 2.0, or FilmArray Torch systems. # I. 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 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). The FilmArray Pneumonia Panel is compatible with BioFire Diagnostics' (BioFire) PCR-based in vitro 5 {5} diagnostic FilmArray, 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 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 singleplex fashion in each well of the array. At the end 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. ### Materials provided in each FilmArray Pneumonia Panel kit: Each kit contains sufficient reagents to test 30 samples (30-test kit; RFIT-ASY-0144) or 6 samples (6-test kit; RFIT-ASY-0145): - Individually-packaged FilmArray Pneumonia Panel pouches - Single-use (1.0 mL) Sample Buffer ampoules 6 {6} - Single-use pre-filled (1.5 mL) Hydration Injection Vials (blue) - Single-use Sample Injection Vials (red) - Individually-packaged Transfer Pipettes **Materials required but not provided:** - FilmArray system including: - FilmArray 2.0 or FilmArrayTouch and accompanying software - FilmArray Pouch Loading Station - 10% bleach solution **Interpretation of Results** When PCR2 is complete, the FilmArray instrument performs a DNA melting analysis on the PCR products and measures the fluorescence signal generated in each well (for more information see appropriate FilmArray Operator's Manual). The FilmArray Software then performs several analyses and assigns a final assay result. The steps in the analyses are described below. Analysis of Melt Curves The FilmArray Software evaluates the DNA melt curve for each well of the PCR2 array to determine if a PCR product was present in that well. If the melt profile indicates the presence of a PCR product, then the analysis software calculates the melting temperature (Tm) of the curve and compares it against the expected Tm range for the assay. If the software determines that the Tm of the curve is within the assay-specific Tm range, the melt curve is called positive. If the software determines that the Tm of the curve is not in the appropriate Tm range, the melt curve is called negative. Analysis of Replicates Once positive melt curves have been identified, the software evaluates the replicates for each assay to determine the assay result. For an assay to be called positive, two associated melt curves must be called positive, and both Tms must be similar. Assays that do not meet these criteria are called negative. Analysis of Assay Results for Bacteria The assays in the FilmArray Pneumonia Panel for detection of bacteria that are reported semi-quantitatively are designed to amplify genes that are present in single copies within the chromosome of the target bacterium and are used to estimate genomic copies of bacterial nucleic acid per milliliter (copies/mL) of specimen. The FilmArray Software calculates an approximate value for each gene target based on real-time PCR amplification data relative to the QSM (internal reference of known quantity). Assays with no measurable 7 {7} amplification or a value below $ 10^{3.5} $ copies/mL are called negative. Assays with a value equal to or greater than $ 10^{3.5} $ copies/mL are called positive. # Organism and Antimicrobial Resistance Gene Interpretation Each positive and negative assay result is interpreted by the FilmArray Software to provide results for the identification of specific bacteria, atypical bacteria, viruses, and antimicrobial resistance (AMR) genes as shown in the Table below. For most analytes detected by the FilmArray Pneumonia Panel, interpretations are based on the result of a single assay. However, results for Staphylococcus aureus, Adenovirus, and the AMR genes require interpretation based on more than one assay result, as discussed in the relevant sections below. | Bacteria – Quantitative Results | Antimicrobial Resistance Genes | | --- | --- | | Acinetobacter calcoaceticus-baumannii complex | blaCTX-M (Extended spectrum beta-lactamase | | Enterobacter cloacae complex | blaIMP (Carbapenem resistance) | | Escherichia coli | blaKPC (Carbapenem resistance) | | Haemophilus influenzae | mecA/mecC and MREJ (Methicillin resistance) | | Klebsiella aerogene | blaNDM (Carbapenem resistance) | | Klebsiella oxytoca | blaOxa48-like (Carbapenem resistance) | | Klebsiella pneumoniae group | blaVIM (Carbapenem resistance) | | Moraxella catarrhalis | Viruses | | 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 | Parainfluenza Virus | | Chlamydia pneumoniae | Respiratory Syncytial Virus | | Legionella pneumophila | | | Mycoplasma pneumoniae | | # Interpretations and Semi-quantitative Bin Results for Bacteria The FilmArray Pneumonia Panel provides a Detected or Not Detected result as well as a semi-quantitative bin result ( $ 10^{4} $ copies/mL, $ 10^{5} $ copies/mL, $ 10^{6} $ copies/mL or $ \geq10^{7} $ copies/mL) for most bacteria. The bin result represents the approximate number of specific bacterial genomes in the specimen and is intended to provide a simple assessment of relative quantities of different bacteria in a lower respiratory specimen based on a molecular method. 8 {8} For bacteria, negative assays (no measurable amplification or value less than 10³·⁵ copies/mL) are reported as Not Detected. Positive assays are reported as Detected and a bin result is assigned based on the assay value. Each bin is defined by discrete upper and lower limits spanning a 1-log range of values (see Table 1) such that the bin result reflects the assay value within the nearest ±0.5-log. Table 1: FilmArray Pneumonia Panel Bin Results for Bacteria | Assay Result | | Reported Result and Bin Result | | | --- | --- | --- | --- | | Negative OR | <10³·⁵ copies/mL | Not Detected | | | Positive AND | ≥10³·⁵ – <10⁴·⁵ copies/mL | Detected | 10⁴ copies/mL | | Positive AND | ≥10⁴·⁵ – <10⁵·⁵ copies/mL | Detected | 10⁵ copies/mL | | Positive AND | ≥10⁵·⁵ – <10⁶·⁵ copies/mL | Detected | 10⁶ copies/mLD | | Positive AND | ≥10⁶·⁵ copies/mL | Detected | ≥10⁷ copies/mL | # Staphylococcus aureus The FilmArray Pneumonia Panel pouch contains two different assays (Saureus1 and Saureus2) for the detection of Staphylococcus aureus. The FilmArray Software interprets each of these assays independently (as described above) and if one or a combination of the assays is positive, the result will be Staphylococcus aureus Detected with the appropriate bin result. If both assays are negative the result will be Staphylococcus aureus Not Detected. # Interpretations for Atypical Bacteria and Viruses Results for most Atypical Bacteria and Viruses are reported qualitatively as Detected or Not Detected based on an individual corresponding assay result. If the assay is positive the result will be Detected, and if the assay is negative, the result will be Not Detected. # Adenovirus The FilmArray Pneumonia Panel pouch contains three different assays (Adenovirus2, Adenovirus3, and Adenovirus7) for the detection of all species and serotypes of Adenovirus. The FilmArray Software interprets each of these assays independently (as described above) and the results are combined as a final result for the virus. If one or any combination of assays is positive, the result will be Adenovirus Detected. If all assays are negative, the result will be Adenovirus Not Detected. 9 {9} Interpretations for Antimicrobial Resistance (AMR) Genes Results for AMR genes are also reported qualitatively (Detected/Not Detected) based on corresponding assays, but only if an applicable bacterium (i.e. potential carriers of the AMR gene; Table 2) is also detected ( $ \geq10^{3.5} $ copies/mL) in the sample. The results for each of the antimicrobial resistance genes will be listed as either: - Detected – when an applicable bacterium is detected AND the antimicrobial resistance gene assay(s) are positive. - Not Detected – when an applicable bacterium is detected AND the antimicrobial resistance gene assay(s) are negative. - N/A – when all applicable bacteria are Not Detected, regardless of the result for the antimicrobial resistance gene assay(s). Table 2: Antimicrobial Resistance (AMR) Genes and Applicable Organisms | AMR Gene Result | Applicable Bacteria | | --- | --- | | mecA/C and MREJ | Staphylococcus aureus | | CTX-MIMPKPCNDMVIM | Acinetobacter calcoaceticus-baumannii complexEnterobacter cloacae complexEscherichia coliKlebsiella aerogenesKlebsiella oxytocaKlebsiella pneumoniae groupProteus spp.Pseudomonas aeruginosaSerratia marcescens | | OXA-48-like | Enterobacter cloacae complexEscherichia coliKlebsiella aerogenesKlebsiella oxytocaKlebsiella pneumoniae groupProteus spp.Serratia marcescens | 10 {10} Each AMR gene result is associated with a single corresponding assay except for the mecA/C and MREJ result, which is dependent on both the mecA/C assay and the MREJ assay. Detection of both Staphylococcus aureus and the mecA/C and MREJ markers is indicative of Methicillin Resistant Staphylococcus Aureus (MRSA). ## Run Information The Run Information section is displayed at the top of both pages of the test report. It provides information about the sample and the run including: Sample ID, Protocol (sample type), pouch information (Pouch Type, Lot Number, and Serial Number), Run Date, Run Status (Completed, Incomplete, Aborted, Instrument Error, Instrument Communication Error, or Software Error), , the identity of the operator who performed the test, and the instrument used to perform the test. Control results are reported as Passed, Failed, or Invalid. The Table 3 below provides additional information for each of the possible control field results. Table 3: Interpretation of Controls Field on the FilmArray Pneumonia Panel Test Report | Control Result | Explanation | Action | | --- | --- | --- | | Passed | The run was successfully completed ANDBoth pouch controls were successful. | NoneReport the results provided on the test report. | | Failed | The run was completed BUTAt least one of the pouch controls (RNA Process Control and/or QSM) failed. | Repeat the test using a new pouch.If the error persists, contact Customer Technical Support for further instruction. | | Invalid | The controls are invalid because the run did not complete.(Typically this indicates a software or hardware error). | Note any error codes displayed during the run and the Run Status field in the Run Information section of the report. Refer to the appropriate FilmArray Operator's Manual or contact Customer Technical Support for further instruction.Once the error is resolved, repeat the test or repeat the test using another instrument. | ## Detection Summary The Detection Summary section is displayed on the first page of the report and lists the Detected results under each category (Bacteria, Antimicrobial Resistance Genes, Atypical Bacteria, and Viruses), including the semi-quantitative ‘Bin (copies/mL)’ results for Bacteria. If there are no Detected results in a specific category, the result shown is Detected: None. 11 {11} ## Result Summary The Results Summary is displayed on the second page of the report and provides a full list of test results for each organism and antimicrobial resistance gene including the 'Bin (copies/mL)' result for Bacteria. Possible results for each organism are Detected, Not Detected, Invalid, and N/A. The Table provides an explanation for each interpretation and any follow-up necessary to obtain a final result. Table 4: Reporting of Results and Required Actions | Result | Explanation | Action | | --- | --- | --- | | Detected | The run was successfully completed AND The pouch controls were successful (Passed) AND The assay(s) for the organism were POSITIVE^{a} | Report results. | | Not Detected | The run was successfully completed AND The pouch controls were successful (Passed) AND The assay(s) for the organism were NEGATIVE^{b} | Report results. | | Invalid | The pouch controls were not successful (Failed) OR The run was not successful (Run Status displayed as: Aborted, Incomplete, Instrument Error or Software Error) | See Table 3 for instruction. | | N/A (Antimicrobial Resistance Genes only) | The run was successfully completed AND The pouch controls were successful (Passed) AND The assay(s) for the organism(s) associated with the antimicrobial resistance gene were NEGATIVE so the results of the antimicrobial resistance gene are not applicable to the test results. | Report results. | $^{a}$ For bacteria, the organism calculated value must be greater than or equal to 10$^{3.5}$ copies/mL for the assay to be POSITIVE. $^{b}$ For bacteria, a NEGATIVE assay result may indicate no amplification or amplification with an organism calculated value less than 10$^{3.5}$ copies/mL ### J. Substantial Equivalence Information: 1. Predicate device name(s): Curetis Unyvero LRT Application 12 {12} 2. Predicate 510(k) number(s): DEN170047 3. Comparison with predicate: | Similarities | | | | --- | --- | --- | | Item | New Device: FilmArray Pneumonia Panel (K180966) | Predicate: Curetis Unyvero LRT Application (DEN170047) | | Indication for 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 10⁴, 10⁵, 10⁶, or ≥10⁷ 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 10⁴, 10⁵, 10⁶, or ≥10⁷ 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 Atypical Bacteria: Chlamydia pneumoniae, Legionella pneumophila, Mycoplasma pneumoniae Antimicrobial Resistance Genes: CTX-M, IMP, KPC, mecA/C + MREJ, NDM, Oxa48-like, VIM Viruses: Adenovirus, Coronavirus, Human Metapneumovirus, Human Rhinovirus/Enterovirus, Influenza A, Influenza B, Parainfluenza virus, Respiratory Syncytial virus 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 | 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. **Microorganisms:** Acinetobacter spp., Chlamydia pneumoniae, Citrobacter freundii, Escherichia coli, Enterobacter cloacae complex, Haemophilus influenzae, Klebsiella oxytoca, Klebsiella pneumoniae, Klebsiella variicola, Legionella pneumophila, Moraxella catarrhalis, Morganella morganii, Mycoplasma pneumoniae, Proteus spp, Pseudomonas aeruginosa, Serratia marcescens, Staphylococcus aureus, Stenotrophomonas maltophilia, Streptococcus pneumoniae **Associated Antimicrobial Resistance Markers:** ctx-M (blaCTX-M, subgroup 1 only), kpc (blaKPC), mecA, ndm (blaNDM), oxa-23 (blaOXA-23), oxa-24 (blaOXA-24), oxa-48 (blaOXA-48), oxa-58 (blaOXA-58), tem (blaTEM), vim (blaVIM) 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. | 13 {13} | Similarities | | | --- | --- | | 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 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 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 the determination of susceptibility or resistance. Due to the genetic similarity between human rhinovirus and | 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. | 14 {14} | Similarities | | | | --- | --- | --- | | | 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. | | | Technological Principles | Multiplex nucleic acid | Same | | Test Interpretation | Automated test interpretation and report generation. | Same | | Controls | Two controls are included in each reagent pouch to control for sample processing and both stages of PCR and melt analysis. | Internal control (a synthetic gene without any homology to known sequences) is processed in every chamber in order to verify the DNA purification, amplification, array hybridization, and detection | | User Complexity | Moderate/Low | Same | | Differences | | | | --- | --- | --- | | Item | New Device: FilmArray Pneumonia Panel (K180966) | Predicate: Curetis Unyvero LRT Application (DEN170047) | | Specimen Types | Sputum-like (induced or expectorated sputum, endotracheal aspirates) and BAL- like (BAL or mini-BAL) specimens | Endotracheal Aspirate specimens | | Analyte | DNA/RNA | DNA only | | Result types | Detection with bin values (in 1-log rounded copies/mL bins) for Bacteria Qualitative for Atypical Bacteria, Antimicrobial Resistance Genes, and Viruses | Qualitative for all analytes | | Instrumentation | FilmArray, FilmArray 2.0, or FilmArray Torch | Unyvero System | | Time to result | About 1 hour | About 4-5 hours | 15 {15} | Differences | | | | --- | --- | --- | | Reagent Storage | Room temperature | Store the Unyvero LRT Cartridge kit at 15°- 25°C. Do not refrigerate. Store the Unyvero Master Mix Tubes frozen at -20°C or below. Thawed Master Mix tubes may be stored at 2-8°C for up to 7 days. Do not re-freeze | # K. Standard/Guidance Document Referenced: - Guidance for Industry and Food and Drug Administration Staff – Highly Multiplexed Microbiological/Medical Countermeasure In Vitro Nucleic Acid Based Diagnostic Devices, (August 27, 2014) - Guidance for Industry and FDA Staff - Class II Special Controls Guidance Document: Respiratory Viral Panel Multiplex Nucleic Acid Assay (October 9, 2009) - Guidance for Industry and FDA Staff - Class II Special Controls Guidance Document: Testing for Detection and Differentiation of Influenza A Virus Subtypes Using Multiplex Nucleic Acid Assays (October 9, 2009) - Guidance for Industry and FDA Staff - Class II Special Controls Guidance Document: Testing for Human Metapneumovirus (hMPV) Using Nucleic Acid Assays (October 9, 2009) - Guidance for Industry and FDA Staff- Statistical Guidance on Reporting Results from Studies Evaluating Diagnostic Tests, (March 13, 2007) - Guidance for Sponsors, Institutional Review Boards, and Food and Drug Administrative Staff – Guidance on Informed Consent for In Vitro Diagnostic Device Studies Using Leftover Human Specimens that are Not Individually Identifiable, (April 25, 2006) - Evaluation of Stability of In Vitro Diagnostic Reagents; Approved Guideline, Clinical and Laboratory Standards Institute (CLSI) – First Edition, EP25-A (September 23, 2009). - Interference Testing in Clinical Chemistry; Approved Guideline, Clinical and Laboratory Standards Institute (CLSI) – Second Addition, EP07-A2 (November, 2005). - Molecular Diagnostic Methods for Infectious Diseases; Approved Guideline, Clinical and Laboratory Standards Institute (CLSI), MM3-A2 (February 2006) - User Protocol for Evaluation of Qualitative Test Performance; Approved Guideline, Clinical and Laboratory Standards Institute (CLSI)– Second Edition, EP12-A2 (January, 2008) - ISO 13485:2016/EN ISO 13485:2016, ‘Medical devices – Quality management systems – Requirements for regulatory purposes’. - ISO 14971:2007 ‘Medical devices – Application of risk management to medical devices’. - EN 13612:2002, ‘Performance evaluation of in vitro diagnostic devices’. - EN 13641:2002, ‘Elimination or reduction of risk of infection related to in vitro diagnostic reagents’. 16 {16} - • EN 62366:2008/IEC 62366-1:2015, 'Medical Devices-Application of usability engineering to medical devices'. - • EN ISO 23640:2015, '*In vitro* diagnostic medical devices – Evaluation of stability of in vitro diagnostic reagents'. - • Guidance for the Content of Premarket Submissions for Software Contained in Medical Devices, FDA Guidance Document (May 11, 2005) - • Guidance for Industry, FDA Reviewers and Compliance on Off-The-Shelf Software Use in Medical Devices (September 9, 1999) - • General Principle of Software Validation; Final Guidance for Industry and FDA Staff (January 11, 2002) - • Content of Premarket Submissions for Management of Cybersecurity in Medical Devices (October 02, 2014) - • Postmarket Management of Cybersecurity in Medical Devices (December 28, 2016) - • ISO 62304:2006, 'Medical device software – Software life-cycle processes' – IEC 62304:2006, November 27, 2008. - • Use of Symbols on Labels and in Labeling of *In Vitro* Diagnostic Devices Intended for Professional Use, FDA Guidance Document (November 30, 2004) - • Guidance for Industry and FDA on Alternative to Certain Prescription Device Labeling Requirements (January 1, 2000) - • ISO 15223-1:2016, 'Medical Devices – Symbols to be used with medical device labels, labeling and information to be supplied – Part 1: General requirements'. - • EN ISO 18113-1:2011, '*In vitro* diagnostic medical devices – Information supplied by the manufacturer (labeling) – Part 1: Terms, definition and general requirements'. - • EN ISO 18113-2:2011, '*In vitro* diagnostic medical devices – Information supplied by the manufacturer (labeling) – Part 2: *In vitro* diagnostic reagents for professional use'. #### **L. Test Principle:** The FilmArray Pneumonia Panel pouch is a closed system disposable that stores all the necessary reagents for sample preparation, reverse transcription, polymerase chain reaction (PCR), and detection in order to isolate, amplify, and detect nucleic acid from multiple lower respiratory pathogens within a single bronchoalveolar lavage (BAL)-like (BAL or mini-BAL) or sputum-like (sputum or ETA) specimen. After sample collection, the user injects hydration solution and sample combined with sample buffer into the pouch, places the pouch into a FilmArray instrument, and starts a run. The entire run process takes about one hour. Additional detail can be found in the appropriate FilmArray Operator's Manual. During a run, the FilmArray system: - Lyses the sample by agitation (bead beading). 17 {17} - Extracts and purifies all nucleic acids from the sample using magnetic bead technology. - Performs nested multiplex PCR by: o First performing reverse transcription and a single, large volume, massively-multiplexed reaction (PCR1) o Then performing multiple singleplex second-stage PCR reactions (PCR2) to amplify sequences within the PCR1 products - Uses endpoint melting curve data to detect and generate a result for each target on the FilmArray Pneumonia Panel array. - For the FilmArray Pneumonia Panel, the system also uses real-time amplification data from the assays relative to a Quantified Standard Material (QSM) included in the pouch to provide an estimated value in genomic copies per milliliter (copies/mL) for bacterial analytes. # **M. Performance Characteristics (if/when applicable):** # 1. Analytical performance: # a. Analytical Sensitivity: A limit of detection (LoD) was established for atypical bacteria and viruses detected by the FilmArray Pneumonia Panel. LoD was estimated by testing dilutions of contrived BAL or sputum samples containing known concentrations of organisms. Confirmation of LoD was achieved by testing at least 20 replicates per samples type on FilmArray, FilmArray 2.0 and FilmArray Torch systems (60 replicates total per sample type). LoD concentration was confirmed when the analyte was detected in at least 95% of the replicates tested. The confirmed LoD for each atypical bacterium or virus detected by the panel (including a LoD for more than one isolate of the more genetically diverse viruses) is listed in Table 5. The LoD concentration is based on quantification of each culture in viable units (TCID50/mL or CFU/mL) and a corresponding molecular LoD concentration (DNA or RNA copies/mL) is provided based on quantitative real-time or digital PCR. 18 {18} Table 5: Summary of Limit of Detection (LoD) for FilmArray Pneumonia Panel Atypical Bacteria and Viruses | Analyte | Isolate Strain/Serotype/Source ID | LoD Concentration$ ^{a} $ | | | --- | --- | --- | --- | | | | Viable Units | Molecular (DNA or RNA) | | Atypical Bacteria | | | | | Chlamydia pneumoniae | TW183ATCC VR-2282 | 5.0E-01 TCID50/mL$ ^b $ | 3.3E+02 copies/mL$ ^b $ | | Legionella pneumophila | Philadelphia-1ATCC 33152 | 5.0E+02 CFU/mL | 1.6E+03 copies/mL | | Mycoplasma pneumoniae | M129Zeptometrix 0801579 | 7.5E+01 TCID50/mL$ ^b $ | 3.5E+03 copies/mL$ ^b $ | | Viruses | | | | | Adenovirus | Species A(A18) ATCC VR-19 | 5.0E+01 TCID50/mL | 9.2E+03 copies/mL | | | Species B (B3)Zeptometrix 0810062CF | 1.0E+00 TCID50/mL | 1.8E+03 copies/mL | | | Species C (C2)ATCC VR-846 | 5.0E+00 TCID50/mL | 7.5E+03 copies/mL | | | Species D (D37)Zeptometrix 0810119CF | 2.5E-01 TCID50/mL$ ^b $ | 2.9E+03 copies/mL$ ^b $ | | | Species E (E4)Zeptometrix 0810070CF | 1.0E-01 TCID50/mL$ ^b $ | 3.5E+04 copies/mL$ ^b $ | | | Species F (F41)ATCC VR-930 | 5.0E+00 TCID50/mL | 5.5E+03 copies/mL | | Coronavirus | 229E | 5.0E-01 TCID50/mL | 8.1E+01 copies/mL | | | HKU1Clinical Specimen$ ^d $ | - | 1.0E+04 copies/mL | | | NL63BEI NR-470 | 2.5E+00 TCID50/mL$ ^c $ | 5.4E+02 copies/mL$ ^c $ | | | OC43ATCC VR-759 | 5.0E+02 TCID50/mL$ ^c $ | 9.3E+03 copies/mL$ ^c $ | | Human Metapneumovirus | 16 Type A1Zeptometrix 0810161CF | 5.0E+01 TCID50/mL | 5.9E+03 copies/mL | | Human Rhinovirus/Enterovirus | Rhinovirus Type 1AZeptometrix 810012CFN | 1.5E+01 TCID50/mL$ ^b $ | 6.6E+03 copies/mL$ ^b $ | | | Echovirus 6Zeptometrix 0810076CF | 1.0E+02 TCID50/mL | 5.7E+02 copies/mL | 19 {19} | Analyte | Isolate Strain/Serotype/Source ID | LoD Concentration^{a} | | | --- | --- | --- | --- | | | | Viable Units | Molecular (DNA or RNA) | | Influenza A | H1N1pdm09 A/SwineNY/03/09 Zeptometrix 0810249CF | 2.5E+00 TCID50/mL^{c} | 1.7E+03 copies/mL^{c} | | | H3N2 A/Port Chalmers/1/73 ATCC VR-810 | 1.0E+00 TCID50/mL^{b} | 2.1E+02 copies/mL^{b} | | Influenza B | B/FL/04/06 Zeptometrix 0810255CF | 5.0E+00 TCID50/mL^{c} | 4.2E+02 copies/mL^{c} | | Parainfluenza Virus | Type 1 Zeptometrix 0810014CF | 2.5E+01 TCID50/mL | 5.2E+03 copies/mL | | | Type 2 Zeptometrix 0810015CF | 2.5E+01 TCID50/mL^{c} | 1.5E+03 copies/mL^{c} | | | Type 3 Zeptometrix 0810016CF | 2.5E+01 TCID50/mL^{c} | 3.8E+02 copies/mL^{c} | | | Type 4A Zeptometrix 0810060CF | 2.5E+02 TCID50/mL | 8.1E+03 copies/mL | | Respiratory Syncytial Virus | Type A Zeptometrix 0810040ACF | 1.0E+00 TCID50/mL | 4.3E+02 copies/mL | $^{a}$ The listed concentration was confirmed with ≥95% detection on each FilmArray system in artificial BAL (aBAL) and/or sputum. $^{b}$ LoD confirmation (≥95% detection) was achieved at a 2 to 5-fold lower concentration in aBAL. $^{c}$ LoD confirmation (≥95% detection) was achieved at a 2 to 5-fold lower concentration in sputum. $^{d}$ No cultured isolates of Coronavirus HKU1 were available for testing. For bacteria, the FilmArray Pneumonia Panel reports a Detected result when the estimated bacterial nucleic acid abundance is ≥10$^{3.5}$ copies/mL, and the panel reports a Not Detected if there is no amplification or the estimated bacterial nucleic acid abundance is <10$^{3.5}$ copies/mL. No assay-specific LoD concentrations were determined for the bacterial analytes, however, each assay was determined to be linear (slope ≈ 1.0 and coefficient of determination (Adj R2) >0.95) and estimates of nucleic acid abundance were determined to be accurate within 0.5 log$_{10}$ copies/mL when compared to an input concentration determined by digital PCR. Antimicrobial resistance (AMR) genes are reported as Detected when an applicable bacterium is detected and the assay for the AMR gene is positive. No AMR gene assay-specific LoD concentrations were determined for the AMR gene, but positive AMR gene assay results were recorded in ≥95% of 90 replicates of applicable bacteria tested at concentrations of 1.0E+04 copies/mL or less in the precision evaluation (see Precision below). 20 {20} # b. Reproducibility: A multisite reproducibility study of the FilmArray Pneumonia Panel was performed with contrived BAL samples over multiples days at three laboratory locations (sites) on a combination of FilmArray, FilmArray 2.0 and FilmArray Torch systems. Testing incorporated a range of potential sources of variability, including run-to-run, day-to-day, and site-to-site variability, which also encompassed different operators, instruments, systems and reagent lots for a total of 30 tests per system and 90 total replicates per sample/concentration. Evaluation of the reproducibility of Detected/Not Detected results for atypical bacteria and viruses included samples containing combinations of five different analytes, at Negative, Low Positive (1×LoD), and Moderate Positive (3×LoD) concentrations. Negative results were obtained from up to 24 additional samples (see evaluation of precision for bacterial analytes below) that were not spiked with the analyte. A summary of results (percent (%) agreement with the expected Detected or Not Detected result) for each atypical bacterium and virus (by site and system) is provided Table 6 below. Table 6: Reproducibility of FilmArray Pneumonia Panel Atypical Bacteria and Virus Results | Analyte | Concentration Tested | Expected Result | Agreement with Expected Result | | | | | --- | --- | --- | --- | --- | --- | --- | | | | | FilmArray | FilmArray 2.0 | FilmArray Torch | All Sites [95% CI] | | | | | Site A | Site B | Site C | | | Atypical Bacteria | | | | | | | | Chlamydia pneumoniae | None(No Analyte) | Not Detected | 780/780100% | 780/780100% | 780/780100% | 2,340/2,340100%[99.8%-100%] | | Legionella pneumophila Philadelphia-1ATCC 33152 | Moderate Positive3× LoD1.5E+03 CFU/mL | Detected | 30/30100% | 30/30100% | 30/30100% | 90/90100%[96.0%-100%] | | | Low Positive1× LoD5.0E+02 CFU/mL | Detected | 30/30100% | 30/30100% | 30/30100% | 90/90100%[96.0%-100%] | | | None(No Analyte) | Not Detected | 720/720100% | 720/720100% | 720/720100% | 2,160/2,160100%[99.8%-100%] | | Mycoplasma pneumoniae | None(No Analyte) | Not Detected | 780/780100% | 780/780100% | 780/780100% | 2,340/2,340100%[99.8%-100%] | | Viruses | | | | | | | 21 {21} | Analyte | Concentration Tested | Expected Result | Agreement with Expected Result | | | | | --- | --- | --- | --- | --- | --- | --- | | | | | FilmArray | FilmArray 2.0 | FilmArray Torch | All Sites [95% CI] | | | | | Site A | Site B | Site C | | | **Adenovirus** Species B Serotype 3 ZeptoMetrix 0810062CF | Moderate Positive 3× LoD 3.0E+00 TCID_{50}/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% [96.0%-100%] | | | Low Positive 1× LoD 1.0E+00 TCID_{50}/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% [96.0%-100%] | | | None (No Analyte) | Not Detected | 720/720 100% | 720/720 100% | 720/720 100% | 2,160/2,160 100% [99.8%-100%] | | **Coronavirus** | None (No Analyte) | Not Detected | 780/780 100% | 776/780 99.5% | 780/780 100% | 2,336/2,340 99.8% [99.6%-100%] | | **Human Metapneumovirus** 16 Type A1 ZeptoMetrix 0810161CF | Moderate Positive 3× LoD 1.5E+02 TCID_{50}/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% [96.0%-100%] | | | Low Positive 1× LoD 5.0E+01 TCID_{50}/mL | Detected | 30/30 100% | 29/30 96.7% | 30/30 100% | 89/90 98.9% [94.0%-100%] | | | None (No Analyte) | Not Detected | 720/720 100% | 720/720 100% | 720/720 100% | 2,160/2,160 100% [99.8%-100%] | | **Human Rhinovirus/Enterovirus** | None (No Analyte) | Not Detected | 779/780 99.9% | 780/780 100% | 779/780 99.9% | 2,338/2,340 99.9% [99.7%-100%] | | **Influenza A** H3N2 A/Port Chalmers/1/73 ATCC VR-810 | Moderate Positive 3× LoD 1.5E+00 TCID_{50}/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% [96.0%-100%] | | | Low Positive 1× LoD 0.5E-01 TCID_{50}/mL | Detected | 30/30 100% | 29/30 96.7% | 30/30 100% | 89/90 98.9% [94.0%-100%] | | | None (No Analyte) | Not Detected | 720/720 100% | 720/720 100% | 720/720 100% | 2,160/2,160 100% [99.8%-100%] | | **Influenza B** | None (No Analyte) | Not Detected | 780/780 100% | 780/780 100% | 780/780 100% | 2,340/2,340 100% [99.8%-100%] | | **Parainfluenza Virus** Type 2 ZeptoMetrix 0810015CF | Moderate Positive 3× LoD 7.5E+01 TCID_{50}/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% [96.0%-100%] | | | Low Positive 1× LoD 2.5E+01 TCID_{50}/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% [96.0%-100%] | 22 {22} | Analyte | Concentration Tested | Expected Result | Agreement with Expected Result | | | | | --- | --- | --- | --- | --- | --- | --- | | | | | FilmArray | FilmArray 2.0 | FilmArray Torch | All Sites [95% CI] | | | | | Site A | Site B | Site C | | | | None (No Analyte) | Not Detected | 720/720 100% | 720/720 100% | 720/720 100% | 2,160/2,160 100% [99.8%-100%] | | Respiratory Syncytial Virus | None (No Analyte) | Not Detected | 780/780 100% | 780/780 100% | 780/780 100% | 2,340/2,340 100% [99.8%-100%] | Precision for bacterial analytes was measured at each concentration as 1) precision of bin results and 2) reproducibility of analyte detection. When a sample containing one or more bacteria is tested repeatedly, the precision of the bin results (probability that each replicate will receive the same bin result) will vary based on the concentration of nucleic acid measured and the relation of that concentration to the limits of each bin. Bin precision may be as low as 50% for values at a bin limit and precision will increase (up to 90% or higher) as the distance of the measured value from a bin limit increases. The precision of FilmArray Pneumonia Panel bin results will follow the model illustrated in Figure 1: - >90% at a bin center (Scenario 1) - ~60 – 90% between a bin limit and bin center (Scenario 2) - ~50% at bin limits (Scenario 3) 23 {23} ![img-0.jpeg](img-0.jpeg) Top: The probability (0.0 - 1.0) of the same bin results for each replicate tested varies based on proximity of the measured value to a bin limit. Bottom: Expected distribution of bin results at different mean measured values. Samples containing bacteria and corresponding antimicrobial (AMR) genes were tested at six different concentrations over the reportable range and below. A summary of the bin precision (percent $(\%)$ of replicates reported in each bin) and the reproducibility of detection is shown at each concentration tested in Table 7 below. Table 7: Reproducibility of FilmArray Pneumonia Panel Bacterial Bin Results on FilmArray, FilmArray 2.0 and FilmArray TorchGrey shading indicates the expected bin results based on the analyte concentration and bold font indicates the bin with the greatest percentage of results at each concentration. | Analyte | Concentration (log10 copies/mL) | % Replicates Reported in Each Bin Result | | | | | Total Detected | | --- | --- | --- | --- | --- | --- | --- | --- | | | | $ \geq 10^7 $ | $ 10^6 $ | $ 10^5 $ | $ 10^4 $ | ND | | | Acinetobacter baumannii (NDM-1) | 7.5 | 90/90(100%) | - | - | - | - | 90/90100% | | | 6.5 | 87/90(96.7%) | 3/90(3.3%) | - | - | - | 90/90100% | 24 {24} | Analyte | Concentration (log10 copies/mL) | % Replicates Reported in Each Bin Result | | | | | Total Detected | | --- | --- | --- | --- | --- | --- | --- | --- | | | | $ \geq 10^7 $ | $ 10^6 $ | $ 10^5 $ | $ 10^4 $ | ND | | | AR-BANK#0033 | 5.5 | - | 82/90(91.1%) | 8/90(8.9%) | - | - | 90/90100% | | | 4.5 | - | 1/90(1.1%) | 80/90(88.9%) | 9/90(10.0%) | - | 90/90100% | | | 3.5 | - | - | 1/90(1.1%) | 74/90(82.2%) | 15/90(16.7%) | 75/9083.3% | | | 2.5 | - | - | - | 1/90(1.1%) | 89/90(98.9%) | 1/901.1% | | | None(No Analyte) | - | - | - | - | 1800/1800(100%) | 0/18000.0% | | Enterobacter aerogenes ATCC 13048 | 7.5 | 90/90(100%) | - | - | - | - | 90/90100% | | | 6.5 | 65/90(72.2%) | 25/90(27.8%) | - | - | - | 90/90100% | | | 5.5 | - | 52/90(57.8%) | 38/90(42.2%) | - | - | 90/90100% | | | 4.5 | - | - | 38/90(42.2%) | 51/90(56.7%) | 1/90(1.1%) | 89/9098.9% | | | 3.5 | - | - | - | 33/90(36.7%) | 57/90(63.3%) | 33/9036.7% | | | 2.5 | - | - | - | 1/90(1.1%) | 89/90(98.9%) | 1/901.1% | | | None(No Analyte) | - | - | - | - | 1800/1800(100%) | 0/18000.0% | | Enterobacter cloacae (VIM) AR-BANK#0154 | 7.0 | 90/90(100%) | - | - | - | - | 90/90100% | | | 6.0 | 4/90(4.4%) | 86/90(95.6%) | - | - | - | 90/90100% | | | 5.0 | - | 6/90(6.7%) | 80/90(88.9%) | - | 4/90(4.4%) | 86/9095.6% | | | 4.0 | - | - | 6/90(6.7%) | 83/90(92.2%) | 1/90(1.1%) | 89/9098.9% | | | 3.0 | - | - | 1/90(1.1%) | 4/90(4.4%) | 85/90(94.4%) | 5/905.6% | | | 2.0 | - | - | - | - | 90/90(100%) | 0/900.0% | | | None (No Analyte) | - | - | - | - | 1800/1800(100%) | 0/18000.0% | 25 {25} | Analyte | Concentration (log10 copies/mL) | % Replicates Reported in Each Bin Result | | | | | Total Detected | | --- | --- | --- | --- | --- | --- | --- | --- | | | | $ \geq 10^7 $ | $ 10^6 $ | $ 10^5 $ | $ 10^4 $ | ND | | | Escherichia coli (IMP) GRE 1062016 | 7.0 | 90/90(100%) | - | - | - | - | 90/90100% | | | 6.0 | 7/90(7.8%) | 82/90(91.1%) | - | - | 1/90 (1.1%) | 89/9098.9% | | | 5.0 | - | 12/90(13.3%) | 78/90(86.7%) | - | - | 90/90100% | | | 4.0 | - | - | 12/90(13.3%) | 77/90(85.6%) | 1/90(1.1%) | 89/9098.9% | | | 3.0 | - | - | 1/90(1.1%) | 15/90(16.7%) | 74/90(82.2%) | 16/9017.8% | | | 2.0 | - | - | - | - | 90/90(100%) | 0/900.0% | | | None (No Analyte) | - | - | - | - | 1800/1800(100%) | 0/18000.0% | | Haemophilus influenzae ATCC 10211 | 7.0 | 89/90(98.9%) | 1/90(1.1%) | - | - | - | 90/90100% | | | 6.0 | 35/90(48.9%) | 55/90(61.1%) | - | - | - | 90/90100% | | | 5.0 | - | 49/90(54.4%) | 40/90(44.4%) | 1/90(1.1%) | - | 90/90100% | | | 4.0 | - | - | 41/90(45.6%) | 49/90(54.4%) | - | 90/90100% | | | 3.0 | - | - | - | 42/90(46.7%) | 48/90(53.3%) | 42/9046.7% | | | 2.0 | - | - | - | - | 90/90(100%) | 0/900.0% | | | None (No Analyte) | - | - | - | - | 1800/1800(100%) | 0/18000.0% | | Klebsiella oxytoca (CTX-M) GRE 1254054 | 7.5 | 90/90(100%) | - | - | - | - | 90/90100% | | | 6.5 | 90/90(100%) | - | - | - | - | 90/90100% | | | 5.5 | 1/90(1.1%) | 84/90(93.3%) | 3/90(3.3%) | - | 2/90(2.2%) | 88/9097.8% | | | 4.5 | - | - | 89/90(98.9%) | - | 1/90(1.1%) | 89/9098.9% | | | 3.5 | - | - | - | 90/90(100%) | - | 90/90100% | 26 {26} | Analyte | Concentration (log10 copies/mL) | % Replicates Reported in Each Bin Result | | | | | Total Detected | | --- | --- | --- | --- | --- | --- | --- | --- | | | | $ \geq 10^7 $ | $ 10^6 $ | $ 10^5 $ | $ 10^4 $ | ND | | | | 2.5 | - | - | 1/90(1.1%) | 1/90(1.1%) | 88/90(97.8%) | 2/902.2% | | | None (No Analyte) | - | - | - | - | 1800/1800(100%) | 0/18000.0% | | Klebsiella pneumoniae(KPC)AR-BANK#0097 | 7.00 | 90/90(100%) | - | - | - | - | 90/90100% | | | 6.00 | 12/90(13.3%) | 78/90(86.7%) | - | - | - | 90/90100% | | | 5.00 | - | 15/90(16.7%) | 75/90(83.3%) | - | - | 90/90100% | | | 4.00 | - | - | 23/90(25.6%) | 66/90(73.3%) | 1/90(1.1%) | 89/9098.9% | | | 3.00 | - | - | - | 15/90(16.7%) | 75/90(83.3%) | 15/9016.7% | | | 2.00 | - | - | - | - | 90/90(100%) | 0/900.0% | | | None (No Analyte) | - | - | - | - | 1800/1800(100%) | 0/18000.0% | | Moraxella catarrhalisATCC 8176 | 7.0 | 90/90(100%) | - | - | - | - | 90/90100% | | | 6.0 | 26/90(28.9%) | 64/90(71.1%) | - | - | - | 90/90100% | | | 5.0 | - | 6/90(6.7%) | 83/90(92.2%) | 1/90(1.1%) | - | 90/90100% | | | 4.0 | - | - | 4/90(4.4%) | 86/90(95.6%) | - | 90/90100% | | | 3.0 | - | - | - | 4/90(4.4%) | 86/90(95.6%) | 4/904.4% | | | 2.0 | - | - | - | - | 90/90(100%) | 0/900.0% | | | None(No Analyte) | - | - | - | - | 1800/1800(100%) | 0/18000.0% | | Proteus mirabilisATCC 35659 | 7.0 | 88/90(97.8%) | - | - | - | 2/90(2.2%) | 88/9097.8% | | | 6.0 | 27/90(30.0%) | 63/90(70.0%) | - | - | - | 90/90100% | | | 5.0 | - | 26/90(28.9%) | 64/90(71.1%) | - | - | 90/90100% | 27 {27} | Analyte | Concentration (log10 copies/mL) | % Replicates Reported in Each Bin Result | | | | | Total Detected | | --- | --- | --- | --- | --- | --- | --- | --- | | | | $ \geq 10^7 $ | $ 10^6 $ | $ 10^5 $ | $ 10^4 $ | ND | | | | 4.0 | - | - | 14/90(15.6%) | 75/90(83.3%) | 1/90(1.1%) | 89/9098.9% | | | 3.0 | - | - | - | 28/90(31.1%) | 62/90(68.9%) | 28/9031.1% | | | 2.0 | - | - | - | - | 90/90(100%) | 0/900.0% | | | None(No Analyte) | - | - | - | - | 1800/1800(100%) | 0/18000.0% | | Pseudomonas aeruginosaATCC 10145 | 7.0 | 90/90(100%) | - | - | - | - | 90/90100% | | | 6.0 | 20/90(22.2%) | 70/90(77.8%) | - | - | - | 90/90100% | | | 5.0 | - | 24/90(26.7%) | 66/90(73.3%) | - | - | 90/90100% | | | 4.0 | - | - | 16/90(17.8%) | 74/90(82.2%) | - | 90/90100% | | | 3.0 | - | - | - | 14/90(15.6%) | 76/90(84.4%) | 14/9015.6% | | | 2.0 | - | - | - | - | 90/90(100%) | 0/900.0% | | | None(No Analyte) | - | - | - | - | 1800/1800(100%) | 0/18000.0% | | Serratia marcescens(OXA-48-like)GRE 1659005 | 7.0 | 90/90(100%) | - | - | - | - | 90/90100% | | | 6.0 | 2/90(2.2%) | 88/90(97.8%) | - | - | - | 90/90100% | | | 5.0 | - | 7/90(7.8%) | 83/90(92.2%) | - | - | 90/90100% | | | 4.0 | - | - | 6/90(6.7%) | 83/90(92.2%) | 1/90(1.1%) | 89/9098.9% | | | 3.0 | - | - | - | 6/90(6.7%) | 84/90(93.3%) | 6/906.7% | | | 2.0 | - | - | - | - | 90/90(100%) | 0/900.0% | | | None(No Analyte) | - | - | - | - | 1800/1800(100%) | 0/18000.0% | 28 {28} | Analyte | Concentration (log10 copies/mL) | % Replicates Reported in Each Bin Result | | | | | Total Detected | | --- | --- | --- | --- | --- | --- | --- | --- | | | | $ \geq 10^7 $ | $ 10^6 $ | $ 10^5 $ | $ 10^4 $ | ND | | | Staphylococcus aureus subsp. aureus (mecA/C and MREJ) ATCC 43300 | 7.0 | 90/90(100%) | - | - | - | - | 90/90100% | | | 6.0 | - | 90/90(100%) | - | - | - | 90/90100% | | | 5.0 | - | - | 90/90(100%) | - | - | 90/90100% | | | 4.0 | - | - | - | 89/90(98.9%) | 1/90(1.1%) | 89/9098.9% | | | 3.0 | - | - | - | - | 90/90(100%) | 0/900.0% | | | 2.0 | - | - | - | - | 90/90(100%) | 0/900.0% | | | None(No Analyte) | - | - | - | 2/1260(0.2%) | 1258/1260(99.8%) | 2/12600.2% | | Streptococcus agalactiae ATCC 13813 | 7.8 | 89/90(98.9%) | 1/90 (1.1%) | - | - | - | 90/90100% | | | 6.8 | 89/90(98.9%) | - | - | - | 1/90 (1.1%) | 89/9098.9% | | | 5.8 | - | 88/90(97.8%) | 1/90(1.1%) | 1/90(1.1%) | - | 90/90100% | | | 4.8 | - | - | 89/90(98.9%) | 1/90(1.1%) | - | 90/90100% | | | 3.8 | - | - | - | 86/90(95.6%) | 4/90(4.4%) | 86/9095.6% | | | 2.8 | - | - | - | 3/90(3.3%) | 87/90(96.7%) | 3/903.3% | | | None(No Analyte) | - | - | - | - | 1800/1800(100%) | 0/18000.0% | | Streptococcus pneumoniae ATCC 6303 | 6.5 | 90/90(100%) | - | - | - | - | 90/90100% | | | 5.5 | - | 90/90(100%) | - | - | - | 90/90100% | | | 4.5 | - | - | 89/90(98.9%) | 1/90(1.1%) | - | 90/90100% | | | 3.5 | - | - | - | 89/90(98.9%) | 1/90(1.1%) | 89/9098.9% | | | 2.5 | - | - | - | - | 90/90(100%) | 0/900.0% | 29 {29} | Analyte | Concentration (log10 copies/mL) | % Replicates Reported in Each Bin Result | | | | | Total Detected | | --- | --- | --- | --- | --- | --- | --- | --- | | | | $ \geq 10^7 $ | $ 10^6 $ | $ 10^5 $ | $ 10^4 $ | ND | | | | 1.5 | - | - | - | - | 90/90(100%) | 0/900.0% | | | None(No Analyte) | - | - | - | - | 1800/1800(100%) | 0/18000.0% | | Streptococcus pyogenesATCC 49399 | 7.8 | 90/90(100%) | - | - | - | - | 90/90100% | | | 6.8 | 90/90(100%) | - | - | - | - | 90/90100% | | | 5.8 | 5/90(5.6%) | 84/90(93.3%) | 1/90(1.1%) | - | - | 90/90100% | | | 4.8 | - | 3/90(3.3%) | 87/90(96.7%) | - | - | 90/90100% | | | 3.8 | - | - | 3/90(3.3%) | 87/90(96.7%) | - | 90/90100% | | | 2.8 | - | - | - | 16/90(18.9%) | 74/90(81.1%) | 16/9017.8% | | | None(No Analyte) | - | - | - | - | 1800/1800(100%) | 0/18000.0% | The precision of the antimicrobial resistance (AMR) genes was measured as the reproducibility of analyte detection on each system and overall. Results are presented in Table 8 as the percent of replicates that are detected at concentrations of the associated bacterium that are within the reportable range, or below the reportable range, as well as the percent agreement with the expected Not Detected result in unspiked samples. 30 {30} **Table 8: Reproducibility of FilmArray Pneumonia Panel Antimicrobial Resistance Gene Results on FilmArray, FilmArray 2.0 and FilmArray Torch** | AMR Gene Organism | Concentration of Organism (copies/mL log10) | Expected Result | Agreement with the Expected Result | | | | | --- | --- | --- | --- | --- | --- | --- | | | | | FilmArray | FilmArray 2.0 | FilmArray Torch | All Systems/Sites [95% CI] | | | | | Site A | Site B | Site C | | | **CTX-M** *Klebsiella oxytoca* **GRE 1254054** | **Reportable Range** | **Detected** | **150/150** 100% | **149/150** 99.3 | **150/150** 100% | **449/450** **99.8%** [98.8%-99.9%] | | | Below Reportable Range | Detected (Variable) | **0/30** 0% | **1/30** 3.3% | **0/30** 0% | **1/90** **1.1%** [0.03%-6.0%] | | | None (No Analyte) | N/A or Not Detected | **600/600** 100% | **599/600** 99.8% | **600/600** 100% | 1799/1800 99.9% [99.7%-100%] | | **IMP** *Escherichia coli* **GRE 1062016** | **Reportable Range** | **Detected** | **120/120** 100% | **120/120** 100% | **120/120** 100% | **360/360** **100%** [99.0%-100%] | | | Below Reportable Range | Detected (Variable) | **10/60** 16.7% | **9/60** 15.0% | **3/60** 5.0% | **22/180** **12.2%** [7.8%-17.9%] | | | None (No Analyte) | N/A or Not Detected | **600/600** 100% | **600/600** 100% | **600/600** 100% | **1800/1800** **100%** [99.8%-100%] | 31 {31} | AMR Gene Organism | Concentration of Organism (copies/mL log10) | Expected Result | Agreement with the Expected Result | | | | | --- | --- | --- | --- | --- | --- | --- | | | | | FilmArray | FilmArray 2.0 | FilmArray Torch | All Systems/Sites [95% CI] | | | | | Site A | Site B | Site C | | | **KPC** *Klebsiella pneumoniae* AR-Bank#0097 | Reportable Range | Detected | 120/120 100% | 119/120 99.2% | 120/120 100% | 359/360 99.7% [98.5%-100%] | | | Below Reportable Range | Detected (Variable) | 14/60 23.3% | 10/60 16.7% | 9/60 15.0% | 33/180 18.3% [13.0%-24.8%] | | | None (No Analyte) | N/A or Not Detected | 600/600 100% | 600/600 100% | 600/600 100% | 1800/1800 100% [99.8%-100%] | | *mecA/C* and MREJ *Staphylococcus aureus* ATCC 43300 | Reportable Range | Detected | 119/120 99.2% | 118/120 98.3% | 120/120 100% | 357/360 99.2% [97.6%-99.8%] | | | Below Reportable Range | Detected (Variable) | 0/60 0% | 0/60 0% | 0/60 0% | 0/180 0% [0.0%-2.0%] | | | None (No Analyte) | N/A or Not Detected | 420/420 100% | 420/420 100% | 420/420 100% | 1260/1260 100% [99.7%-100%] | | **NDM** *Acinetobacter baumannii* AR-Bank#0033 | Reportable Range | Detected | 150/150 100% | 149/150 99.3% | 150/150 100% | 449/450 99.8% [98.8%-100%] | | | Below Reportable Range | Detected (Variable) | 1/30 3.3% | 1/30 3.3% | 0/30 0% | 2/90 2.2% [0.3%-7.8%] | | | None (No Analyte) | N/A or Not Detected | 599/600 99.8% | 600/600 100% | 600/600 100% | 1799/1800 99.9% [99.7%-100%] | | **OXA-48-like** *Serratia marcescens* GRE 1659005 | Reportable Range | Detected | 120/120 100% | 119/120 99.2% | 120/120 100% | 359/360 99.70% [98.5%-100%] | | | Below Reportable Range | Detected (Variable) | 14/60 23.3% | 12/60 20.0% | 9/60 15.0% | 35/180 19.4% [13.9%-26.0%] | | | None (No Analyte) | N/A or Not Detected | 598/600 99.7% | 600/600 100% | 600/600 100% | 1798/1800 99.9% [99.6%-100%] | | **VIM** | Reportable Range | Detected | 120/120 100% | 120/120 100% | 120/120 100% | 360/360 100% | 32 {32} | AMR Gene Organism | Concentration of Organism (copies/mL log10) | Expected Result | Agreement with the Expected Result | | | | | --- | --- | --- | --- | --- | --- | --- | | | | | FilmArray | FilmArray 2.0 | FilmArray Torch | All Systems/Sites [95% CI] | | | | | Site A | Site B | Site C | | | Enterobacter cloacae AR-BANK#0154 | | | | | | [99.0%-100%] | | | Below Reportable Range | Detected (Variable) | 10/60 16.7% | 9/60 15.0% | 3/60 5.0% | 22/180 12.2% [7.8%-17.9%] | | | None (No Analyte) | N/A or Not Detected | 599/600 99.8% | 600/600 100% | 600/600 100% | 1799/1800 99.9% [99.7%-100%] | #### c. Linearity/assay reportable range: ##### Linearity Linearity of the FilmArray Pneumonia Panel was performed to evaluate unbinned value response to changes in concentration and to determine if an offset exists between the unbinned value and a dPCR (digital PCR) determined molecular concentration (the unbinned value is calculated using real-time amplification data for the analyte relative to a known standard (Quantified Standard Material or QSM)). A series of contrived samples were prepared in artificial bronchoalveolar lavage (aBAL) matrix such that each bacterial analyte was tested at six different concentrations spanning 1.5 – 7.8 log10 (copies/mL). The input concentration of each cultured bacterium was determined by digital PCR (dPCR), which served as the reference concentrations for offset determination. Once prepared, each sample was tested repeatedly at each concentration on a single day for a total of eighteen replicate measures per analyte. The unbinned values generated by the FilmArray Pneumonia Panel assays were plotted as a function of the nominal (dPCR) input concentrations to determine the linearity of the response. All plot data were observed to be linear and this was confirmed through mathematical modeling. For each analyte, data bounding the reporting range ( $ \sim3.0-7.0\log10 $ (copies/mL)) were fit using linear and quadratic models. In all cases, the linear fit model was preferred as it was sufficient to describe the change in unbinned values as a function of changing inputs and the addition of curvature did not improve the fit predictions. The best fit linear model coefficients for each bacterial FilmArray Pneumonia Panel assay were evaluated (slope, y-intercept, and coefficient of determination (Adj R$ ^{2} $)). The slope provides a direct measure of how the unbinned value responds to changes in input concentration, where a slope near 1.0 demonstrates that the unbinned values change in direct proportion to the input concentration. The best fit slopes of the assays varied within the range of 0.975 – 1.114, demonstrating that changes in the 33 {33} unbinned values display a ~1:1 relationship to changes in the input concentration. The y-intercept from the linear equation represents the theoretical offset predicted to exist at a zero input concentration. The Adj R² value provides a measure of the goodness of fit between the linear model and the data, where values of 1.0 indicate perfect agreement and values of 0 indicate random association displaying no correlation. The best fit Adj R² values of the assays were high, ranging from 0.951 – 0.989, further demonstrating strong agreement between the data and the linear model. The FilmArray Pneumonia Panel unbinned value was shown to respond proportionately to changes in input concentration following a linear relationship defined by slopes around 1.0. The offset of each assay was found to be within the range 0.01 – 0.27-log10 (copies/mL) and does not change substantially as a function of concentration over the reportable range. These data demonstrate that the Pneumonia Panel unbinned value is accurate and thus that the FilmArray Pneumonia Panel reported bin results will accurately reflect the concentration of analytes in a test sample. d. Traceability, Stability, Expected values (controls, calibrators, or methods): # Process Controls Two process controls are included in each pouch: RNA Process Control: The RNA Process Control assay targets an RNA transcript from the yeast Schizosaccharomyces pombe. The yeast is present in the pouch in a freeze-dried form and becomes rehydrated when sample is loaded. The control material is carried through all stages of the test process, including lysis, nucleic acid purification, reverse transcription, PCR1, dilution, PCR2, and DNA melting. A positive RNA Process Control result indicates that all steps carried out in the FilmArray Pneumonia Panel pouch were successful. Quantified Standard Material (QSM) Control: The QSM assay detects a quantified standard synthetic nucleic acid that is subject to all stages of the test process following sample lysis (bead beating). A positive QSM control result indicates that the expected level of QSM is present (approximately 10⁶ copies/mL) for use in determining assay and bin results for bacterial analytes. Both control assays must be positive for the test run to pass. If the controls fail, the sample should be retested using a new pouch. 34 {34} ## External Controls External controls are not provided with the FilmArray Pneumonia Panel. However, five frozen (-70°C) external control mixes (ECMs) (see Table below) were provided to the study sites for daily testing during the prospective clinical trial and the clinical study testing contrived specimens. FilmArray operators were required to complete a valid ECM run (correct results obtained) on each day of clinical specimen testing (tested on a rotating basis). Table 9: External Control Mixes (EC's) Utilized in the Clinical Evaluations | External Control Mixes | Expected Calls | | --- | --- | | ECM1 | Acinetobacter calcoaceticus-baumannii complex, Aspergillus spp., Adenovirus, Influenza A, Respiratory Syncytial Virus, Chlamydophila pneumoniae, Legionella pneumophila, Proteus spp, blaNDM (Carbapenem resistance), blaKPC (Carbapenem resistance) | | ECM2 | Enterobacter aerogenes/cloacae complex, Cryptococcus neoformans/gattii, Coronavirus, Middle Eastern Respiratory Syndrome Coronavirus, Haemophilus influenzae, Moraxella catarrhalis, Pseudomonas aeruginosa, Streptococcus pneumoniae, blaCTX-M (Extended spectrum beta-lactamase), blaOXA48-like (Carbapenem resistance) | | ECM3 | Klebsiella pneumoniae group, Escherichia coli, Human Metapneumovirus, Influenza B, Klebsiella oxytoca Serratia marcescens, Staphylococcus aureus, blaVIM (Carbapenem resistance), blaIMP (Carbapenem resistance) | | ECM4 | Staphylococcus aureus, Stenotrophomonas maltophilia Pneumocystis jirovecii, Human Rhinovirus/Enterovirus Parainfluenza Virus, Streptococcus agalactiae, Mycoplasma pneumoniae, Streptococcus pyogenes mecA/mecC and MREJ (Methicillin resistance) | | ECM5 | None | External controls should be used in accordance with laboratory protocols and the appropriate accrediting organization requirements, as applicable. Molecular grade water or saline can be used as an external negative control. Previously characterized positive samples or negative samples spiked with well characterized organisms can be used as external positive controls. ## Specimen Stability A study was performed to assess and validate specimen storage and handling recommendations for BAL-like and sputum-like specimens that will be tested with the FilmArray Pneumonia Panel. Contrived samples were prepared with representative panel organism spiked into pools of residual 35 {35} clinical BAL or sputum specimens. One sample (QL) was composed of spiked RNA virus, DNA virus, and atypical bacteria at 3x LoD, as well as the organisms that were native to the pooled clinical specimen matrix. The other sample (QT) was composed of gram-positive and gram-negative bacteria (including fastidious species and isolates carting antimicrobial resistance genes) spiked at 10⁴ copies/mL. Samples were tested on the FilmArray Pneumonia Panel immediately after sample preparation to generate control data (no storage; D0) and were then stored in refrigerated conditions for additional testing after one (D1) and two (D2) days, based on current lower respiratory specimen storage guidelines provided by the CDC. Ten replicates were tested at each storage timepoint and results were compared to the control. In the contrived BAL and sputum samples, detection of analytes was consistent between the stored and unstored samples out to two days of refrigerated storage (9/10 or 10/10 detected result), with no noticeable changes in amplification data associated with storage. Only the mecA/C and MREJ result in the BAL sample was detected in fewer replicates than anticipated on the second day of storage (mecA/C and MREJ was reported as Detected in 7/9 replicates (77.8%) with a Staphylococcus aureus Detected result), but review of the amplification data did not suggest a difference in amplification or template levels in the stored sample compared to the unstored samples. Detection of native analytes (viruses, bacteria, and AMR genes) within the same contrived BAL and sputum samples was also consistent between the stored and unstored samples out to two days of refrigerated storage. Results are presented in the Table below. Table 10: FilmArray Pneumonia Panel Results for Stored Contrived BAL and Sputum Samples | Analyte | Strain | Concentration | Analyte Detection | | | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | | | BAL | | | Sputum | | | | | | | No Storage (D0) | Day 1 (D1) | Day 2 (D2) | No Storage (D0) | Day 1 (D1) | Day 2 (D2) | | **Spiked Analytes** | | | | | | | | | | Adenovirus (Type B3) | Zeptometrix 0810062CF | 3.0E+00 TCID₅₀/mL | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | | Parainfluenza Virus (Type 4a) | Zeptometrix 0810060CF | 7.5E+02 TCID₅₀/mL | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | | Respiratory Syncytial Virus | Zeptometrix 0810040ACF | 3.0E+00 TCID₅₀/mL | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | | Legionella pneumophila | ATCC 33152 | 1.5E+03 CFU/mL | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 36 {36} | Analyte | Strain | Concentration | Analyte Detection | | | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | | | BAL | | | Sputum | | | | | | | No Storage (D0) | Day 1 (D1) | Day 2 (D2) | No Storage (D0) | Day 1 (D1) | Day 2 (D2) | | Haemophilus influenzae | ATCC 10211 | 1.0E+04 copies/mL | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | | Klebsiella oxytoca | GRE 1254054 | | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | | CTX-M | | | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | | Proteus mirabilis | ATCC 35659 | | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | | Staphylococcus aureus | ATCC 43300 | | 10/10 | 10/10 | 9/10 | 10/10 | 10/10 | 10/10 | | mecA/C & MREJ | | | 10/10 | 10/10 | 7/9 | 10/10 | 10/10 | 10/10 | | Streptococcus pneumoniae | ATCC 6303 | | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | | NativeAnalytes | | | | | | | | | | Human Rhinovirus/Enterovirus$ ^a $ | Unknown | 10/10 | 9/10 | 10/10 | 9/10 | 10/10 | 10/10 | | | Influenza A | | | - | | | 10/10 | 10/10 | 10/10 | | Enterobacter cloacae complex$ ^a $ | | | 1…

FilmArray Pneumonia Panel

Device Facts

Real-World Evidence

Clinical Evidence

AI Performance

Indications for Use

Device Story

Clinical Evidence

Technological Characteristics

Indications for Use

Regulatory Classification

Identification

Special Controls

Predicate Devices

Submission Summary (Full Text)

Panel 1

Section Matches

Product Code Matches

Special Control Matches

My Alerts

FilmArray Pneumonia Panel

Device Facts

Real-World Evidence

Clinical Evidence

AI Performance

Indications for Use

Device Story

Clinical Evidence

Technological Characteristics

Indications for Use

Regulatory Classification

Identification

Special Controls

Predicate Devices

Submission Summary (Full Text)

Panel 1

Section Matches

Product Code Matches

Special Control Matches

My Alerts

Create Alert

Search Filters

Agent Token

Welcome

Check your email

Welcome!