Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The description details a PCR-based nucleic acid test with automated interpretation of melt curve analysis. There is no mention of AI or ML algorithms being used for data analysis or interpretation.

Therapeutic

No.
The document indicates that the FilmArray® Respiratory Panel 2 (RP2) is a diagnostic test designed for the detection and identification of respiratory viral and bacterial nucleic acids. It is explicitly stated that "The results of this test should not be used as the sole basis for diagnosis, treatment, or other patient management decisions," and its purpose is to "aid in the diagnosis of respiratory infection." A therapeutic device is used for treatment or therapy, not solely for diagnosis.

Diagnostic

Yes

The "Intended Use" section states that the device aids in the "diagnosis of respiratory infection" by detecting and identifying specific viral and bacterial nucleic acids.

SaMD (Software as a Medical Device)

No

The device description clearly outlines a system that includes hardware components (FilmArray 2.0 or FilmArray Torch systems, pouches with reagents, instruments with bladders, seals, pistons, Peltier devices, digital camera) in addition to the software module. The software is integral to the operation of the hardware and the analysis of the data generated by the hardware.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

Here's why:

Intended Use: The "Intended Use / Indications for Use" section explicitly states that the FilmArray® Respiratory Panel 2 (RP2) is a "multiplexed nucleic acid test intended for use with FilmArray® 2.0 or FilmArray® Torch systems for the simultaneous qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swabs (NPS) obtained from individuals suspected of respiratory tract infections." This clearly describes a test performed in vitro (outside the body) on a biological specimen (nasopharyngeal swabs) to provide diagnostic information.
Device Description: The description details the process of performing the test on a biological sample within the FilmArray system, including nucleic acid extraction, PCR amplification, and detection of specific targets. This is characteristic of an in vitro diagnostic device.
Clinical and Analytical Performance Studies: The document describes clinical and analytical studies conducted to evaluate the performance of the device in detecting the target pathogens in clinical specimens. This is a requirement for IVD devices to demonstrate their accuracy and reliability.
Predicate Device: The mention of a "Predicate Device(s)" (K160068 - FilmArray Respiratory Panel (RP)) further confirms its classification as an IVD, as predicate devices are used for comparison in the regulatory submission process for new IVDs.

The entire document describes a device designed to be used in a laboratory setting to analyze patient samples for diagnostic purposes, which is the definition of an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The FilmArray® Respiratory Panel 2 (RP2) is a multiplexed nucleic acid test intended for use with FilmArray® 2.0 or FilmArray® Torch systems for the simultaneous qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swabs (NPS) obtained from individuals suspected of respiratory tract infections. The following organism types and subtypes are identified using the FilmArray® RP2:

Adenovirus
Coronavirus 229E
Coronavirus HKU1
Coronavirus NL63
Coronavirus OC43
Human Metapneumovirus
Human Rhinovirus/Enterovirus
Influenza A, including subtypes H1, H1-2009, and H3
Influenza B
Parainfluenza Virus 1
Parainfluenza Virus 2
Parainfluenza Virus 3
Parainfluenza Virus 4
Respiratory Syncytial Virus
Bordetella parapertussis (IS1001)
Bordetella pertussis (ptxP)
Chlamydia pneumoniae
Mycoplasma pneumoniae

The detection and identification of specific viral and bacterial nucleic acids from individuals exhibiting signs and/or symptoms of a respiratory infection aids in the diagnosis of 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, or lower respiratory tract infection that may not be detected by a nasopharyngeal swab specimen. Positive results do not rule out co-infection with other organisms: the agent(s) detected by the FilmArray® RP2 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 respiratory tract infection.

Due to the genetic similarity between Human Rhinovirus and Enterovirus, the FilmArray® RP2 cannot reliably differentiate them. A positive FilmArray® RP2 Rhinovirus/Enterovirus result should be followed up using an alternate method (e.g., cell culture or sequence analysis) if differentiation is required.

Performance characteristics for Influenza A were established when Influenza A H1-2009 and A H3 were the predominant Influenza A viruses in circulation. Performance of detecting Influenza A may vary if other Influenza A strains are circulating or a novel Influenza A virus emerges. 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.

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

OCC, OEM, OEP, OOU, OTG, OZE, OZX, OZY, OZZ, OOI, NSU

Device Description

The FilmArray Respiratory Panel 2 (RP2) is designed to simultaneously identify 21 different potential pathogens of respiratory tract infection from a single NPS specimen in a time frame (~45 minutes) that allows the test results to be used in determining appropriate patient treatment and management. FilmArray RP2 is compatible with BioFire Diagnostics' (BioFire) PCR-based in vitro diagnostic FilmArray 2.0 and FilmArray Torch systems for infectious disease testing. A specific software module (i.e. FilmArray RP2 pouch module) is used to perform FilmArray RP2 testing on these systems.

A test is initiated by loading Hydration Solution into one port of the FilmArray pouch and an NPS sample (in transport media) mixed with the provided Sample Buffer into the other port of the FilmArray RP2 pouch and placing it in a FilmArray instrument. The pouch contains all of the reagents required for specimen testing and analysis in a freeze-dried format; the addition of Hydration Solution and Sample/Buffer Mix rehydrates the reagents. After the pouch is prepared, the FilmArray Software guides the user though the steps of placing the pouch into the instrument, scanning the pouch barcode, entering the sample identification, and initiating the run.

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

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

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

Mentions image processing

A digital camera placed in front of the 2nd stage PCR captures fluorescent images of the PCR reactions and software interprets the data.

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

nasopharyngeal swabs (NPS)

Indicated Patient Age Range

Not Found

Intended User / Care Setting

Prescription Use (Part 21 CFR 801 Subpart D)

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

Not Found

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

Clinical Performance Study:

Sample Size: 1635 residual NPS specimens in viral transport media (VTM) were initially acquired. 23 specimens were excluded due to non-compliance, resulting in a final data set of 1612 prospective specimens.
- Category I (fresh): 940 specimens, collected and tested fresh from September 2016.
- Category II (archived/frozen): 695 specimens, collected between January and March 2016 and immediately frozen for later testing.
Data Source: Multi-center study conducted at three geographically distinct U.S. study sites during portions of the 2015-2016 and 2016-2017 respiratory illness seasons.
Annotation Protocol: Performance was evaluated by comparing FilmArray RP2 test results with those from an FDA-cleared multiplexed respiratory pathogen panel (main comparator method) and two analytically-validated PCR assays followed by bi-directional sequencing for B. parapertussis. Discrepant results were further investigated using independent molecular methods and/or comparator method retesting.

Preselected Archived Specimens Study:

Sample Size: 217 preselected archived retrospective clinical specimens were initially received.
- Group 1: 197 specimens (containing analytes on the FDA-cleared multiplexed respiratory pathogens panel comparator method). 3 specimens excluded from analysis, resulting in 194 valid specimens.
- Group 2: 20 specimens (containing B. parapertussis). All 20 specimens yielded valid results.
- Total valid archived specimens: 214.
Data Source: Archived NPS in VTM specimens from BioFire. Selected because they previously tested positive for specific low-prevalence analytes.
Annotation Protocol: Composition/integrity of specimens confirmed with confirmatory molecular methods (PCR followed by bi-directional sequencing for B. parapertussis or an FDA-cleared multiplexed respiratory pathogen panel). All positive archived specimens not confirmed by the respective comparator method were excluded from performance calculation.

Contrived Specimens Study:

Sample Size: 50 contrived positive specimens (at 2x LoD and other concentrations spanning the clinically relevant range) and 50 un-spiked negative specimens.
Data Source: Prepared using individual unique residual NPS specimens that had previously tested negative by the FDA-cleared multiplexed respiratory pathogen panel. Spiking performed using multiple quantified isolates of Influenza A H1.
Annotation Protocol: Contrived positive specimens were prepared and randomized along with negative specimens such that the analyte status was unknown to users.

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

Clinical Performance Study:

Study Type: Multi-center, prospective clinical study.
Sample Size: 1612 prospective specimens (940 fresh, 672 archived/frozen).
Key Results:
- Positive Percent Agreement (PPA): Ranged from 66.7% (B. pertussis, Overall) to 100% (various analytes).
- Negative Percent Agreement (NPA): Ranged from 90.2% (HRV/EV, Fresh) to 100% (various analytes).
- Overall success rate for initial specimen tests: 99.3% (1611/1623).
- Instrument success rate: 99.9% (1621/1623).
- Pouch control success rate: 99.4% (1611/1621).

Testing of Preselected Archived Specimens:

Study Type: Retrospective evaluation of preselected archived specimens.
Sample Size: 214 valid archived specimens (194 from Group 1, 20 from Group 2).
Key Results:
- Positive Percent Agreement (PPA): Ranged from 94.7% (HRV/EV) to 100% (most analytes).
- Negative Percent Agreement (NPA): Ranged from 96.0% (HRV/EV) to 100% (most analytes).

Testing of Contrived Specimens:

Study Type: Evaluation of contrived specimens.
Sample Size: 48 positive contrived specimens for Influenza A H1 (at various multiples of LoD) and 50 negative specimens.
Key Results:
- Positive Percent Agreement: 97.9% (47/48) overall for Influenza A H1.
- Negative Percent Agreement: 100% (50/50).

Selected Analytical Studies:

Limit of Detection (LoD): Determined for all analytes on both FilmArray 2.0 and FilmArray Torch systems by testing 20 replicates at various concentrations. LoD was confirmed when at least 19/20 replicates (95%) were detected.
Inclusivity: 177 isolates representing temporal and geographic diversity were tested. All isolates were detected at concentrations within 10x LoD. In silico analysis also used.
Exclusivity: Evaluated for non-specific amplification with 22 on-panel organisms and 50 off-panel organisms (including near-neighbors and normal flora) tested at high concentrations (typically ≥1x10^6 CFU/mL for bacteria/fungi and ≥1x10^4 TCID50/mL for viruses, representing 70-400,000 fold higher than LoD). Predicted and observed cross-reactivity noted for Bordetella species and Influenza A subtypes of swine origin.
Interference: Evaluated various endogenous substances (blood, mucus, human genomic DNA), competitive microorganisms, exogenous substances (medications, washes), and specimen collection materials. None showed interference except for bleach, which damaged organisms/nucleic acids leading to inaccurate results.
Reproducibility: Performed at three sites with a combination of FilmArray 2.0 and FilmArray Torch systems, varying operators, systems, and pouch lots. Samples included negative, low positive (1x LoD), and moderate positive (3x LoD) concentrations. 480 total valid runs for each analyte group. Agreement with expected results generally very high (e.g., 99.6% overall). Melting temperature (Tm) reproducibility showed standard deviation of +/- 0.5C or less.

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

Positive Percent Agreement (PPA) for each analyte was calculated as 100% x (TP / (TP + FN)). Negative Percent Agreement (NPA) was calculated as 100% x (TN / (TN + FP)).
(Specific values are provided in the tables within the document for each analyte and study type.)

Selected examples from Clinical Performance Summary (Table 4):

Adenovirus: PPA 94.6% (70/74), NPA 96.9% (1490/1538)
CoV-229E: PPA 91.7% (11/12), NPA 99.7% (1595/1600)
Human Rhinovirus/Enterovirus: PPA 97.5% (425/436), NPA 93.5% (1099/1176)
Influenza A: PPA 100% (78/78), NPA 100% (1531/1531)
RSV: PPA 99.4% (175/176), NPA 98.3% (1412/1436)
B. parapertussis (IS1001): PPA 85.7% (6/7), NPA 100% (1605/1605)
M. pneumoniae: PPA 95.8% (23/24), NPA 99.7% (1583/1588)

Selected examples from Archived Specimen Performance Data Summary (Table 8):

CoV-229E: PPA 100% (15/15), NPA 100% (175/175)
Influenza A: PPA 100% (22/22), NPA 100% (172/172)
Bordetella parapertussis (IS1001): PPA 100% (16/16), NPA 100% (4/4)

Influenza A H1 Performance Using Contrived Specimens (Table 9):

Positive Percent Agreement (Combined): 97.9% (47/48)
Negative Percent Agreement: 100% (50/50)

Reproducibility (Table 26):

Overall Agreement with the Expected Result All Analytes and All Test Levels: 99.6% (9562/9600) (95% Confidence Interval: 99.5% - 99.7%)

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

K160068

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

Not Found

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

Not Found

Regulation Number and Section

§ 866.3980 Respiratory viral panel multiplex nucleic acid assay.

(a)
Identification. A respiratory viral panel multiplex nucleic acid assay is a qualitative in vitro diagnostic device intended to simultaneously detect and identify multiple viral nucleic acids extracted from human respiratory specimens or viral culture. The detection and identification of a specific viral nucleic acid from individuals exhibiting signs and symptoms of respiratory infection aids in the diagnosis of respiratory viral infection when used in conjunction with other clinical and laboratory findings. The device is intended for detection and identification of a combination of the following viruses:(1) Influenza A and Influenza B;
(2) Influenza A subtype H1 and Influenza A subtype H3;
(3) Respiratory Syncytial Virus subtype A and Respiratory Syncytial Virus subtype B;
(4) Parainfluenza 1, Parainfluenza 2, and Parainfluenza 3 virus;
(5) Human Metapneumovirus;
(6) Rhinovirus; and
(7) Adenovirus.
(b)
Classification. Class II (special controls). The special controls are:(1) FDA's guidance document entitled “Class II Special Controls Guidance Document: Respiratory Viral Panel Multiplex Nucleic Acid Assay;”
(2) For a device that detects and identifies Human Metapneumovirus, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Human Metapneumovirus (hMPV) Using Nucleic Acid Assays;” and
(3) For a device that detects and differentiates Influenza A subtype H1 and subtype H3, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Detection and Differentiation of Influenza A Virus Subtypes Using Multiplex Nucleic Acid Assays.” See § 866.1(e) for the availability of these guidance documents.

Summary

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Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular seal with the words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter. Inside the circle is an abstract symbol that resembles a stylized human figure or a caduceus, with three intertwined shapes representing people.

Public Health Service

May 30, 2017

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

BioFire Diagnostics, LLC Kristen J. Kanack, Ph.D. Vice President, Regulatory and Clinical Affairs 515 Colorow Drive Salt Lake City, UT 84108

Re: K170604

Trade/Device Name: FilmArray® Respiratory Panel 2 (RP2) Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory Viral Panel Multiplex Nucleic Acid Assay Regulatory Class: II Product Code: OCC, OEM, OEP, OOU, OTG, OZE, OZX, OZY, OZZ, OOI, NSU Dated: February 28, 2017 Received: March 1, 2017

Dear Dr. Kanack:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food. Drug. and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

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

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

1

If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address

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

http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

Steven R. Gitterman -S

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

Enclosure

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Indications for Use

510(k) Number (if known) K170604

Device Name

The FilmArray® Respiratory Panel 2 (RP2)

Indications for Use (Describe)

The FilmArray® Respiratory Panel 2 (RP2) is a multiplexed nucleic acid test intended for use with FilmArray® 2.0 or FilmArray® Torch systems for the simultaneous qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swabs (NPS) obtained from individuals suspected of respiratory tract infections. The following organism types and subtypes are identified using the FilmArray® RP2:

· Adenovirus
Coronavirus 229E
· Coronavirus HKU1
· Coronavirus NL63
Coronavirus OC43
Human Metapneumovirus
Human Rhinovirus/Enterovirus
· Influenza A, including subtypes H1, H1-2009, and H3
Influenza B
Parainfluenza Virus 1
Parainfluenza Virus 2
Parainfluenza Virus 3
Parainfluenza Virus 4
· Respiratory Syncytial Virus

3

· Bordetella parapertussis (IS1001)
Bordetella pertussis (ptxP)
Chlamydia pneumoniae
Mycoplasma pneumoniae

The detection and identification of specific viral and bacterial nucleic acids from individuals exhibiting signs and/or symptoms of a respiratory infection aids in the diagnosis of 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, or lower respiratory tract infection that may not be detected by a nasopharyngeal swab specimen. Positive results do not rule out co-infection with other organisms: the agent(s) detected by the FilmArray® RP2 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 respiratory tract infection.

Due to the genetic similarity between Human Rhinovirus and Enterovirus, the FilmArray® RP2 cannot reliably differentiate them. A positive FilmArray® RP2 Rhinovirus/Enterovirus result should be followed up using an alternate method (e.g., cell culture or sequence analysis) if differentiation is required.

Performance characteristics for Influenza A were established when Influenza A H1-2009 and A H3 were the predominant Influenza A viruses in circulation. Performance of detecting Influenza A may vary if other Influenza A strains are circulating or a novel Influenza A virus emerges. 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.

Type of Use (Select one or both, as applicable)
Prescription Use (Part 21 CFR 801 Subpart D)	Over-The-Counter Use (21 CFR 801 Subpart C)

CONTINUE ON A SEPARATE PAGE IF NEEDED.

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DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.

The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:

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"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."

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510(k) Summary BioFire Diagnostics, LLC

FilmArray Respiratory Panel 2 (RP2)

Introduction:

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

Submitted by:

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

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

Contact: Kristen J. Kanack, ext. 330

Date Submitted: February 28, 2017

Device Name and Classification:

Trade Name: FilmArray Respiratory Panel 2 (RP2)

Regulation Number: 21 CFR 866.3980

Classification Name: Respiratory viral panel multiplex nucleic acid assay

Predicate Device:

K160068 - FilmArray Respiratory Panel (RP)

Intended Use:

The FilmArray® Respiratory Panel 2 (RP2) is a multiplexed nucleic acid test intended for use with FilmArray® 2.0 or FilmArray® Torch systems for the simultaneous qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swabs (NPS) obtained from individuals suspected of respiratory tract infections. The following organism types and subtypes are identified using the FilmArray RP2:

Adenovirus ●
Coronavirus 229E ●
Coronavirus HKU1
Coronavirus NL63
Coronavirus OC43

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Human Metapneumovirus ●
Human Rhinovirus/Enterovirus
Influenza A, including subtypes H1, H1-2009, and H3
Influenza B
. Parainfluenza Virus 1
Parainfluenza Virus 2
Parainfluenza Virus 3
. Parainfluenza Virus 4
Respiratory Syncytial Virus
Bordetella parapertussis (IS 1001)
Bordetella pertussis (ptxP)
Chlamydia pneumoniae ●
Mycoplasma pneumoniae ●

The detection and identification of specific viral and bacterial nucleic acids from individuals exhibiting signs and/or symptoms of a respiratory infection aids in the diagnosis of 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, or lower respiratory tract infection that may not be detected by a nasopharyngeal swab specimen. Positive results do not rule out co-infection with other organisms: the agent(s) detected by the FilmArray RP2 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 respiratory tract infection.

Due to the genetic similarity between Human Rhinovirus and Enterovirus, the FilmArray RP2 cannot reliably differentiate them. A positive FilmArray RP2 Rhinovirus/Enterovirus result should be followed up using an alternate method (e.g., cell culture or sequence analysis) if differentiation is required.

Performance characteristics for Influenza A were established when Influenza A H1-2009, A H1, and A H3 were the predominant Influenza A viruses in circulation. Performance of detecting Influenza A may vary if other Influenza A strains are circulating or a novel Influenza A virus emerges. 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.

Device Description:

The FilmArray Respiratory Panel 2 (RP2) is designed to simultaneously identify 21 different potential pathogens (see Table 1) of respiratory tract infection from a single NPS specimen in a

7

time frame (~45 minutes) that allows the test results to be used in determining appropriate patient treatment and management. FilmArray RP2 is compatible with BioFire Diagnostics' (BioFire) PCR-based in vitro diagnostic FilmArray 2.0 and FilmArray Torch systems for infectious disease testing. A specific software module (i.e. FilmArray RP2 pouch module) is used to perform FilmArray RP2 testing on these systems.

Viral Targets Detected	Bacterial Targets Detected
Adenovirus
Coronavirus 229E
Coronavirus HKU1
Coronavirus NL63
Coronavirus OC43
Human Metapneumovirus
Human Rhinovirus/Enterovirus
Influenza A, including subtypes
H1, H3 and H1-2009
Influenza B
Parainfluenza Virus 1
Parainfluenza Virus 2
Parainfluenza Virus 3
Parainfluenza Virus 4
Respiratory Syncytial Virus	Bordetella parapertussis (IS1001)
Bordetella pertussis (ptxP)
Chlamydia pneumoniae
Mycoplasma pneumoniae

Table 1. Pathogens Detected by the FilmArray RP2

A test is initiated by loading Hydration Solution into one port of the FilmArray pouch and an NPS sample (in transport media) mixed with the provided Sample Buffer into the other port of the FilmArray RP2 pouch and placing it in a FilmArray instrument. The pouch contains all of the reagents required for specimen testing and analysis in a freeze-dried format; the addition of Hydration Solution and Sample/Buffer Mix rehydrates the reagents. After the pouch is prepared, the FilmArray Software guides the user though the steps of placing the pouch into the instrument, scanning the pouch barcode, entering the sample identification, and initiating the run.

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

Nucleic acid extraction occurs within the FilmArray pouch using mechanical and chemical lysis followed by purification using standard magnetic bead technology. After extracting and purifying nucleic acids from the unprocessed sample, the FilmArray performs a nested multiplex PCR that is executed in two stages. During the first stage, the FilmArray performs a single, large volume, highly multiplexed reverse transcription PCR (rt-PCR) reaction. The products from first

8

stage PCR are then diluted and combined with a fresh, primer-free master mix and a fluorescent double stranded DNA binding dye (LC Green® Plus, BioFire Diagnostics). The solution is then distributed to each well of the array. Array wells contain sets of primers designed specifically to amplify sequences internal to the PCR products generated during the first stage PCR reaction. The 2nd stage PCR, or nested PCR, is performed in singleplex fashion in each well of the array. At the conclusion of the 2nd stage PCR, the array is interrogated by melt curve analysis for the detection of signature amplicons denoting the presence of specific targets. A digital camera placed in front of the 2nd stage PCR captures fluorescent images of the PCR reactions and software interprets the data.

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

Substantial Equivalence:

The FilmArray Respiratory Panel 2 is substantially equivalent to the FilmArray Respiratory Panel (K160068), which was most recently cleared on February 8, 2016 and determined to be a Class II device.

Table 2 compares the FilmArray Respiratory Panel 2 (RP2) to the FilmArray Respiratory Panel (RP) and outlines the similarities and differences between the two tests.

| Element | New Device:
FilmArray Respiratory Panel 2 (RP2) | Predicate:
FilmArray Respiratory Panel (RP)
(K160068) |
|-----------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------|
| Specimen Types | NPS in VTM | Same |
| Organisms
Detected | Adenovirus, Coronavirus 229E, Coronavirus
HKU1, Coronavirus NL63, Coronavirus OC43,
Human Metapneumovirus, Influenza A,
Influenza A subtype H1, Influenza A subtype
H3, Influenza A subtype H1-2009, Influenza B,
Parainfluenza Virus 1, Parainfluenza Virus 2,
Parainfluenza Virus 3, Parainfluenza Virus 4,
Human Rhinovirus/Enterovirus, Respiratory
Syncytial Virus, Bordetella parapertussis,
Bordetella pertussis, Chlamydia pneumoniae,
and Mycoplasma pneumoniae | Does not detect Bordetella parapertussis
Reports Chlamydia pneumoniae by alternate
name: Chlamydophila pneumoniae |
| Analyte | DNA/RNA | Same |
| Technological
Principles | Multiplex nucleic acid | Same |
| Instrumentation | FilmArray 2.0 or FilmArray Torch | FilmArray, FilmArray 2.0, or FilmArray Torch |
| Time to result | About 45 minutes | About 1 hour |
| Reagent Storage | Room temperature | Same |
| Test
Interpretation | Automated test interpretation and report
generation. User cannot access raw data. | Same |

Table 2. Comparison of the FilmArray Respiratory Panel 2 (RP2) and the FilmArray Respiratory Panel (RP)

9

| Controls | Two controls are included in each reagent pouch
to control for sample processing and both stages
of PCR and melt analysis. | Same |
|-----------------|----------------------------------------------------------------------------------------------------------------------------------|------|
| User Complexity | Moderate/Low | Same |

Summary of Performance Data

Clinical Performance

The clinical performance of the FilmArray RP2 was established during a multi-center study conducted at three geographically distinct U.S. study sites during portions of the 2015-2016 and 2016-2017 respiratory illness seasons. A total of 1635 residual NPS specimens in viral transport media (VTM) were acquired for the prospective clinical study. Between January and March 2016. specimens were prospectively collected from all comers meeting the study eligibility criteria and immediately frozen (N=695 specimens) for later testing as prospective archived/frozen (Category II) specimens. Between September 2016, specimens were prospectively collected from all comers meeting the study eligibility criteria and tested fresh (N=940 specimens) as prospective fresh (Category I) specimens. Category II specimens were distributed to study sites beginning in September 2016. Study sites also began testing Category I specimens at this time. At each site, Category II specimens were thawed and tested according to the study procedures as time permitted over the remaining duration of the clinical study. A total of 23 prospective specimens (Category I and II specimens) were excluded from the final performance data analysis due to incompliance with the study protocol. The most common reasons for specimen exclusion were that a valid external control was not completed on the day of testing, that specimens were tested outside the 3-day refrigerated storage window, or that the specimen was found to not meet the inclusion criteria after the specimen had been enrolled. The final data set consisted of 1612 prospective specimens. Table 3 provides a summary of demographic information for the 1612 specimens included in the prospective study.

		Overall
	Male	867
(54%)	331
(57%)	271
(51%)	265
(53%)
Sex	Female	745
(46%)	250
(43%)	256
(49%)	239
(47%)
Age	IS1001 )	A747
Zeptometrix 0801461	$4.1E+01$ CFU/mL
$6.0E+01$ IS1001 copies/mLc	20/20
100%	19/20
95%
Bordetella pertussis (ptxP)	A639
Zeptometrix 0801459	$1.0E+03$ CFU/mL	20/20
100%	20/20
100%
Chlamydia pneumoniae	TW183
ATCC VR-2282	$1.0E-01$ TCID50/mL
$6.6E+01$ copies/mL	20/20
100%	19/20
95%
Mycoplasma pneumoniae	M129
Zeptometrix 0801579	$1.0E+01$ TCID50/mL
$4.6E+02$ copies/mLd	20/20
100%	20/20
100%

"The differences in LoD concentration between Adenovines species (5.0E-02 - 1.0E-01 TCD-J/mL) or between Human Rhinovirus (1.0E-01 - 3.0E+02TCID=ymL) may not indicate actual different species or serotypes, but ather the variability of the TCD-measurement method.

" A cultured isolate of Coronavirus HKU1 was not available for testing. LoD for Coronavirus HKU1 was theremined by testing dilutions of a clinical NPS specimen known to contain the virus. The anount of viral RNA copies/mL) was determined by real-ime RT-PCR against a standard curve.

IS/001 sequences can be present in more than one copy per cell, so the relationship between CFU/mL and copies/mL may vary from strain and culture to culture. LoD was determined based on the copy number of IS 001 measured by an independent quantitative real-ime PCR assay and for this culture.

The copy number of Mycoplasma pneumoniae was determined by a multicopy assay (16S rRNA) and may not accurately reflect the number of copies detected by the single-copy FilmArray RP2 gene target.

Note: LoD concentrations of the cultured viruses and obligate intracelleular bacteria (C. pneumoniae and M. pneumoniae) are provided in units of TCID-6 (50% Tissue Culture Infectious Dose). TCID50 is an indirect measure of viral or bacterial concentration based on infectivity and cytotoxicity and will therefore vary considerably depending on technique and methodology (including cell type, culture media and conditions, cytotoxicity of the virus, etc.). It is not appropriate to make determinations on relative sensitivity of different molecular assays for detection of viruses and bacteria based on LoD values measured in TCID50/mL

Inclusivity

Analytical reactivity (inclusivity) was evaluated with a collection of 177 isolates that represent temporal and geographic diversity of FilmArray RP2 analytes, including relevant species, strains, serotypes, or genotypes. All isolates tested were detected by the FilmArray RP2 at concentrations within 10× LoD. When possible, in silico analysis of sequence data was used to make predictions of assay reactivity for less common strains or serotypes that were not tested but that may be detected by the FilmArray RP2 (Table 11 - Table 22).

20

RP2 influenza A assays will react variably with non-human influenza A viruses and rarely encountered human influenza A viruses that are not H1, H1-2009 or H3; generally producing Influenza A Equivocal or Influenza A (no subtype detected) results.

Note: FilmArray RP2 Influenza A (subtype) and Influenza B assays are predicted to react with attenuated viruses used in vaccines.

| Speciesa | Serotype | Isolate ID/Source | [Strain/Location/Year] | xLoD
Dectected | Result |
|----------|----------|----------------------------------------|------------------------------------|-------------------|------------------------|
| A | 12 | ATCC VR-863 | [Huie/Massachusetts] | 3x | |
| A | 18 | ATCC VR-19 | [Washington DC/1954] | 1x | |
| A | 31 | Zeptometrix 0810073CF | - | 3x | |
| A | 3 | Zeptometrix 0810062CF | - | 3x | |
| A | 7A | Zeptometrix 0810021CF | - | 1x | |
| B | 7d/d2 | Univ of Iowa Research Foundation | [Iowa/2001] | 3x | |
| B | 7h | Univ of Iowa Research Foundation | [Iowa/1999] | 3x | |
| B | 11 | Univ of Iowa Research Foundation | [Wisconsin/2005] | 3x | |
| B | 14 | Univ of Iowa Research Foundation | [Missouri/2005] | 3x | |
| B | 16 | ATCC VR-17 | [CH.79/Saudia Arabia/1955] | 3x | |
| B | 21 | Univ of Iowa Research Foundation | [Missouri/2005] | 3x | |
| B | 34 | ATCC VR-716 | [Compton/1972] | 3x | |
| B | 35 | ATCC VR-718 | [Holden] | 3x | |
| B | 50 | ATCC VR-1602 | [Wan/Amsterdam/1988] | 3x | Adenovirus
Detected |
| C | 1 | Zeptometrix 0810050CF | - | 3x | |
| C | 2 | ATCC VR-846 | [Adenoid 6] | 1x | |
| C | 5 | Zeptometrix 0810020CF | - | 3x | |
| C | 6 | ATCC VR-6 | [Tonsil 99/Washington DC] | 3x | |
| D | 8 | Zeptometrix 0810069CF | - | 3x | |
| D | 20 | Zeptometrix 0810115CF | - | 3x | |
| D | 37 | Zeptometrix 0810119CF | - | 1x | |
| E | 4a | Univ of Iowa Research Foundation | [S Carolina/2004] | 1x | |
| E | 4 | Zeptometrix 0810070CF | - | 3x | |
| E | 40 | Zeptometrix 0810084CF
NCPV 0101141v | - | 3x | |
| F | 41 | ATCC VR-930 | [Tak/73-
3544/Netherlands/1973] | 1x | |
| F | 41 | Zeptometrix 0810085CF | - | 3x | |

Table 11. Adenovirus Isolates Tested and Detected by FilmArray RP2

ª In silco analysis of available sequences predicts that the FilmArray RP2 will also react with Adenovirus B55, C57, species D serotypes, and G52.

Table 12. Coronavirus Isolates/Specimens Tested and Detected by FilmArray RP2
-------------------------------------------------------------------------------

| Coronavirus
Type | Isolate ID/Source | [Location/Year] | xLoD
Dectected | Result |
|---------------------|-----------------------|--------------------|-------------------|------------------|
| 229E | ATCC VR-740 | - | 1x | Coronavirus 229E |
| 229E | Zeptometrix 0810229CF | - | 3x | Coronavirus 229E |
| HKU1 | Clinical Specimen | [Utah/2015] | 1x | |
| HKU1 | Clinical Specimen | [Utah/2015] | 3x | |
| HKU1 | Clinical Specimen | [Utah/2015] | 3x | Coronavirus HKU1 |
| HKU1 | Clinical Specimen | [S. Carolina/2010] | 3x | Coronavirus HKU1 |
| HKU1 | Clinical Specimen | [Detroit/2010] | 3x | Coronavirus HKU1 |
| NL63 | BEI NR-470a | [Amsterdam/2003] | 1x | Coronavirus NL63 |
| NL63 | Zeptometrix 0810228CF | - | 3x | Coronavirus NL63 |
| OC43 | ATCC VR-759b | - | 1x | Coronavirus OC43 |
| OC43 | Zeptometrix 0810024CF | - | 3x | Coronavirus OC43 |

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

21

b Discontinued part number; see ATCC VR-1558.

| Genotype | Serotype | Isolate ID/Source | [Location/Year] | xLoD
Detected | Result |
|----------|----------|----------------------------------|-----------------|------------------|--------------------------|
| A1 | 16 | Zeptometrix 0810161CF | [Iowa10/2003] | 1x | Human
Metapneumovirus |
| A1 | 9 | Zeptometrix 0810160CF | [Iowa3/2002] | 3x | |
| A2 | 20 | Zeptometrix 0810163CF | [Iowa14/2003] | 3x | |
| A2 | 27 | Zeptometrix 0810164CF | [Iowa27/2004] | 3x | |
| B1 | 3 | Zeptometrix 0810156CF | [Peru2/2002] | 3x | |
| B1 | 5 | Zeptometrix 0810158CF | [Peru3/2003] | 3x | |
| B1 | 13 | Univ of Iowa Research Foundation | [Iowa7/2003] | 3x | |
| B2 | 4 | Zeptometrix 0810157CF | [Peru1/2002] | 3x | |
| | 8 | Zeptometrix 0810159CF | [Peru6/2003] | 3x | |
| | 18 | Zeptometrix 0810162CF | [Iowa18/2003] | 3x | |
| | 22 | Univ of Iowa Research Foundation | [Iowa16/2003] | 3x | |

Table 13. Human Metapneumovirus Isolates Tested and Detected by FilmArray RP2

Table 14. Human Rhinovirus and Enterovirus Isolates Tested and Detected by FilmArray RP2

| Species | Serotype Detected | Result |------------------|--- | Human Rhinovirus | | | 1 | | 2 | | 7 | A | 16 DC/1960] | | 34 | | 57 Rhinovirus/
Enterovirus |
| | 77 | | 85 | | 3 | | 14 | B | 17 | | 27 | | 42 | | 83 | Enterovirus | | A | | Rhinovirus/
Enterovirus |
| | | B | | | C 2] | C | D | 68 | Type Detected | | H1N1 | Human H1 |
| | | | | | | | | | | | Swine | | | | | | Swine | | | H1N2 A/Rockefeller | H1N1
pdm09 | Human H1-2009 |
| | | | | | | | | | | | | | | | | | Human | | | | | | H3 |
| H3N2g | | | | | | | | | H3N2v | Human | H2N2 | Human (no subtype
detected) |
| (no subtype
detected) |
| H2N3 | Equivocal |
| H5N1 | Avian | H5N2 | pintail/Washington/40964/2014] | H5N3 | detected) |
| H5N8 | | H7N7 | | H7N9 | | H10N7 | Equivocal |
| Lineage Detected | N/A | | | | | | Victoria | | | Yamagata | | | | Isolate ID/Source | [Strain/Location/Year] | xLOD
|
--------------------|-------------------------|---------------------------------------------------------------------------|------------------|-----------------------------------------|
| | | | |
| Zeptometrix 0810012CFN | [1A] | 1x | |
| ATCC VR-482 | [HGP] | 3x | |
| ATCC VR-1601 | [68-CV11] | 3x | |
| ATCC VR-283 | [11757/Washington
| 3x | |
| ATCC VR-507ª | [137-3] | 3x | |
| ATCC VR-1600 | [Ch47] | 3x | Human
| ATCC VR-1187 | [130-63] | 3x | |
| ATCC VR-1195 | [50-525-CV54] | 3x | |
| ATCC VR-483 | [FEB] | 3x | |
| ATCC VR-284 | [1059/S Carolina/1959] | 3x | |
| ATCC VR-1663 | [33342/N Carolina/1959] | 3x | |
| ATCC VR-1137 | [5870] | 3x | |
| ATCC VR-338 | [56822] | 3x | |
| ATCC VR-1193 | [Baylor 7] | 3x | |
| | | | |
| Coxsackievirus 10 | ATCC VR-168 | [NY/1950] | 3x | |
| Enterovirus 71 | ATCC VR-1432 | [H] | 3x | |
| Coxsackievirus A9 | Zeptometrix 0810017CF | - | 3x | Human
| Coxsackievirus B3 | Zeptometrix 0810074CF | - | 3x | |
| Coxsackievirus B4 | Zeptometrix 0810075CF | - | 3x | |
| Echovirus 6 | Zeptometrix 0810076CF | - | 3x | |
| Echovirus 9 | Zeptometrix 0810077CF | - | 3x | |
| Echovirus 11 | Zeptometrix 0810023CF | - | 3x | |
| Coxsackievirus A21 | ATCC VR-850 | [Kuykendall/California/195
| 3x | |
| Coxsackievirus A24 | ATCC VR-583 | [DN-19/Texas/1963] | 3x | |
| ATCC VR-1823 | [US/MO/2014-18947] | 1x | |
| Isolate ID/Source | [Strain/Location/Year] | xLoD
| Result | |
| Zeptometrix 0810036CF | [New Caledonia/20/1999] | 1x | | |
| ATCC VR-219 | [NWS/1933] | 3x | Influenza A
| ATCC VR-95 | [PR/8/1934] | 10xa | |
| ATCC VR-96 | [Weiss/1943] | 3x | |
| ATCC VR-97 | [FM/1/1947] | 3x | |
| ATCC VR-98 | [Mal/302/1954] | 3x | |
| ATCC VR-546 | [Denver/1/1957] | 3x | |
| Zeptometrix 0810036CFN | [Solomon Isl/03/2006] | 3x | |
| Zeptometrix 0810244CF | [Brisbane/59/2007] | 3x | |
| ATCC VR-333 | [A/Swine/Iowa/15/1930] | 3x | |
| ATCC VR-99 | [A/Swine/1976/1931] | 3x | |
| ATCC VR-897 | [A/New Jersey/8/76 (Hsw1N1)] | 10xa | |
| Recombinant | BEI NR-9677b | [Kilbourne F63, A/NWS/1934 (HA) x
Institute/5/1957 (NA)] | 3x | |
| Zeptometrix 0810249CFN | [SwineNY/03/2009] | 1x | Influenza A
| Zeptometrix 0810248CFN | [SwineNY/01/2009] | 3x | |
| Zeptometrix 0810109CFN | [SwineNY/02/2009] | 3x | |
| Zeptometrix 0810109CFJ | [Canada/6294/2009] | 3x | |
| Zeptometrix 0810165CF | [California/07/2009] | 3x | |
| Zeptometrix 0810166CF | [Mexico/4108/2009] | 3x | |
| BEI NR-19823c | [Netherlands/2629/2009] | 3x | |
| BEI NR-44345d | [Hong Kong/H090-761-V1(0)/2009] | 10xe | |
| BEI NR-42938f | [Georgia/F32551/2012] | 3x | |
| ATCC VR-810 | [Port Chalmers/1/1973] | 1x | |
| ATCC VR-776 | [Alice (live attenuated vaccine)] | 3x | |
| Zeptometrix 0810238CF | [Texas/50/2012] | 3x | |
| ATCC VR-547 | [Aichi/2/1968] | 3x | Influenza A
| ATCC VR-544 | [Hong Kong/8/1968] | 3x | |
| ATCC VR-822 | [Victoria/3/1975] | 3x | |
| Zeptometrix 0810252CF | [Wisconsin/67/2005] | 3x | |
| Zeptometrix 0810138CF | [Brisbane/10/2007] | 3x | |
| Recombinant | ATCC VR-777 | [MCR2(A/England/42/72xA/PR8/34)] | 3x | |
| Clinical Specimen | [Ohio/2012] | 3x | |
| BEI NR-2775h | [Japan/305/1957] | 10xe | Influenza A
| Recombinant | BEI NR-9679i | [Korea/426/1968xPuerto Rico/8/1934 ] | 10xe | Influenza A
| MRI Globalj | [Mallard/Alberta/79/2003] | 3x | Influenza A
| MRI Globalj | [A/Chicken/Yunnan/1251/2003] | 3x | |
| MRI Globalj | [A/Northern
| 3x | Influenza A |
| BEI NR-9682k | [A/Duck/Singapore/645/1997] | 3x | (no subtype
| MRI Globalj | [A/Gryfalcon/Washington/41088-6/2014] | 3x | |
| MRI Globalj | [A/Netherlands/219/2003] | 3x | |
| MRI Globalj | [A/Anhui/01/2013] | 3x | |
| BEI NR-2765l | [Chicken/Germany/N/49] | 3x | Influenza A
| Isolate ID/Source | [Strain/Location/Year] | xLoD
| Result | |
| ATCC VR-101 | [Lee/1940] | 3x | | |
| ATCC VR-102 | [Allen/1945] | 3x | | |
| ATCC VR-103 | [GL/1739/1954] | 3x | | |
| ATCC VR-296 | [1/Maryland/1959] | 3x | | |
| ATCC VR-295 | [2/Taiwan/1962] | 3x | | |
| ATCC VR-786 | [Brigit/Russia/1969] | 3x | | |
| ATCC VR-823 | [5/Hong Kong/1972] | 3x | Influenza B | |
| Zeptometrix 0810258CF | [2506/Malaysia/2004] | 3x | | |
| CDC 2005743348 | [1/Ohio/2005] | 3x | | |
| Zeptometrix 0810256CF | [07/Florida/2004] | 3x | | |
| Zeptometrix 0810255CF | [04/Florida/2006] | 1x | | |
| Zeptometrix 0810241CF | [1/Wisconsin/2010] | 3x | | |
| Zeptometrix 0810239CF | [2/Massachusetts/2012] | 3x | | |

ª Discontinued part number; see ATCC VR-1365.

Table 15. Influenza A Isolates Tested and Detected by FilmArray RP2

22

4 Reported as Influenza A (no subtype detected) at 3× LoD.

b Genomic RNA obtained through the NIH Biodefense and Emerging Infections Resources Respiratory NAID, NIH Kilbourne F63: A/NWS/1934 (HA) x A/Rockefeller Institute/5/1957 (NA) (H1N2), Reassortant NWS-F, NR-9677.

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

4 Organism obtained through BEI Resources, NIAID, NIH: Influenza A Virus, A/Hong Kong/H090-761-V (0)/2009 (H1N1)pdm09, NR-44345. e Reported as Influenza A Equivocal or Influenza A (no subtype detected) at 3× LoD.

f Organism obtained through BEI Resources, NIAID, NIH: Influenza A Virus, A/Georgia/F32551/2012 (HIN1)pdm09, NR-42938.

8 Human isolates of recent swine variant H3N2v) were not available for testing, but reactivity near LoD is predicted by sequence analysis.

4 Genomic RNA obtained through the NIH Biodefense and Emerging Infections Resources Repository, NIAID, NIH: Genomic RNA from Influenza A Virus, A/Japan/305/1957 (H2N2), NR-2775.

23

1 Genomic RNA obtained through the NIH Biodefense and Emerging Infections Resources Repository, NIAID, NIH: Genomic RNA from Kilbourne F38: A/Korea/426/1968 (HA, NA) x A/Puerto Rico/8/1934 (H2N2), NR-9679.

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

Genomic RNA obtained through the NIH Biodefense and Emerging Infections Research Resources Repository NIAID, NIH: Genomic RNA from Kilbourne F181: A/duck/Singapore/645/1997 (H5N3), Wild Type, NR-9682.

1 Genomic RNA obtained through the NIH Biodefense and Emerging Infections Resources Repository NIAID, NIH: Genomic RNA from Influenza A Virus, A/chicken/Germany/N/1949 (H10N7), NR-2765

Table 16. Influenza B Isolates Tested and Detected by FilmArray RP2

Table 17. Parainfluenza Virus Isolates Tested and Detected by FilmArray RP2

| Type | Subtype | Isolate ID/Source | [Strain/Location/Year] | xLoD
Detected | Result |
|------|---------|------------------------|----------------------------------|------------------|--------------------------|
| 1 | | Zeptometrix 0810014CF | - | 1x | Parainfluenza Virus
1 |
| | | ATCC VR-94 | [C-35/Washington DC/1957] | 3x | |
| | | BEI NR-3226a | [C39] | 3x | |
| | | BEI NR-48680b | [FRA/29221106/2009] | 3x | |
| 2 | | Zeptometrix 0810015CF | - | 1x | Parainfluenza Virus
2 |
| | | ATCC VR-92 | [Greer/Ohio/1955] | 3x | |
| 3 | | Zeptometrix 0810016CF | - | 1x | Parainfluenza Virus
3 |
| | | ATCC VR-93 | [C-243/Washington DC/1957] | 3x | |
| | | BEI NR-3233c | [NIH 47885, Wash/47885/57] | 3x | |
| 4 | A | Zeptometrix 0810060CF | - | 1x | Parainfluenza Virus
4 |
| | | ATCC VR-1378 | [M-25/1958] | 3x | |
| | B | Zeptometrix 0810060BCF | - | 3x | |
| | | ATCC VR-1377 | [CH-19503/Washington
DC/1962] | 3x | |

ª Discontinued part number.

Obtained through BEI Resources, NIAID, NIH: Human Parainfluenza Virus 1, HPV 1/FRA/29221106/2009, NR-48680.

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

	Table 18. Respiratory Syncytial Virus Isolates Tested and Detected by FilmArray RP2

| Type | Source | [Strain/Location/Year] | xLoD
Detected | Result |
|------|------------------------|----------------------------|------------------|--------------------------------|
| A | Zeptometrix 0810040ACF | [2006] | 1x | |
| A | ATCC VR-26 | [Long/Maryland/1956] | 3x | |
| A | ATCC VR-1540 | [A2/Melbourne/1961] | 3x | Respiratory
Syncytial Virus |
| B | Zeptometrix 0810040CF | [Ch-93 (18)-18] | 3x | |
| B | ATCC VR-1400 | [WV/14617/1985] | 3x | |
| B | ATCC VR-955 | [9320/Massachusetts/1977] | 3x | |
| B | ATCC VR-1580 | [18537/Washington DC/1962] | 10x | |

24

| Species | Source | [Strain/Location/Year] | xLoD
Detected | Result |
|--------------------------------------------------|---------------------|--------------------------------|------------------|-----------------------------------------|
| Bordetella parapertussis | Zeptometrix 0801461 | [A747] | 1x | |
| Bordetella parapertussis | Zeptometrix 0801462 | [E595] | 3x | |
| Bordetella parapertussis | ATCC 15237 | [NCTC 10853] | 3x | Bordetella
parapertussis
(IS1001) |
| Bordetella parapertussis | ATCC 15311 | [NCTC 5952] | 3x | Bordetella
parapertussis
(IS1001) |
| Bordetella parapertussis | ATCC BAA-587 | [12822/Germany/1993] | 3x | Bordetella
parapertussis
(IS1001) |
| Bordetella bronchiseptica
(containing IS1001) | NRRL B-59909 | [MBORD849/
Pig/Netherlands] | 3x | |

Table 19. Bordetella parapertussis (and Bordetella bronchiseptica) Isolates Tested and Detected by FilmArray RP2

2 Readivity with IS1001 sequences in B. bronchiseptica reactivity of the assay, however the analyte will be inacurately reported as B. parapertussis. The assay does not react with IS 1001-like sequences in B. holmesil (see Analytical Reactivity).

Table 20. Bordetella pertussis Isolates Tested and Detected by FilmArray RP2

Isolate ID/Source	[Strain]	xLoD Detected	Result
Zeptometrix 0801459	[A639]	1x
Zeptometrix 0801460	[E431]	3x
ATCC 8467	[F]	3x
ATCC 9340	[5,17921]	3x
ATCC 9797	[18323/NCTC 10739]	3x	Bordetella
ATCC 10380	[10-536]	3x	pertussis (ptxP)
ATCC 51445	[CNCTC Hp 12/63,623]	3x
ATCC BAA-589	[Tohama]	3x
ATCC BAA-1335	[MN2531]	3x

Table 21. Chlamydia pneumoniae Isolates Tested and Detected by FilmArray RP2

Isolate ID/Source	[Strain/Location/Year]	xLoD Detected	Result
ATCC VR-2282	[TW-183/Taiwan/1965]	1x	Chlamydia
pneumoniae
ATCC VR-1310	[CWL-029]	3x
ATCC VR-1360	[CM-1/Georgia]	3x
ATCC 53592	[AR-39/Seattle/1983]	3x

	Table 22. Mycoplasma pneumoniae Isolates Tested and Detected by FilmArray RP2
--	--------------------------------------------------------------------------------------	--	--

Type	Isolate ID/Source	[Strain]	xLoD Detected	Result
1	Zeptometrix 0801579	[M129]	1x	Mycoplasma
pneumoniae
	ATCC 29342	[M129-B7]	3x
	ATCC 29085	[PI 1428]	3x
2	ATCC 15531	[FH strain of Eaton Agent [NCTC 10119]	3x
	ATCC 15492	[Mac]	3x
unknown	ATCC 15293	[M52]	3x
	ATCC 15377	[Bru]	3x
	ATCC 39505	[Mutant 22]	3x
		ATCC 49894	[UTMB-10P]	3x

Exclusivity

The potential for non-specific amplification and detection by the FilmArray RP2 assays was evaluated in silico by sequence analysis and testing of high concentrations of organisms in contrived samples. A total of 22 on-panel organisms were tested to assess the potential for intrapanel cross-reactivity and 50 off-panel organisms were tested to evaluate panel specificity. Offpanel organisms included normal respiratory flora and pathogens that may be present in NPS

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specimens as well as near-neighbors or species genetically related to the organisms detected by the FilmArray RP2. The on-panel and off-panel organisms tested are shown in Table 23.

The final concentration of analyte in the sample (typically at least 1×106 CFU/mL for bacteria and fungi and at least 1×10 TCID0.05 by Fisher's exact test) and therefore do not increatent variance in precision of the FilmArray RP2 Bordeella parapertusis (IS1001) results.

The reproducibility (standard deviation) of melting temperature (Tm) for the amplification products generated by the FilmArray RP2 was also evaluated, with a Tm standard deviation for

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each assay of ± 0.5℃ or less observed within and between the FilmArray 2.0 and FilmArray Torch systems (Table 27).

		Site A		Site C		Site B		Site C		All Sites/Systems
Analyte	Assay	Mean	StDev	Mean	StDev	Mean	StDev	Mean	StDev	Mean	StDev
Controls	yeastRNA	82.3	± 0.3	82.1	± 0.2	82.2	± 0.3	82.0	± 0.2	82.1	± 0.3
	PCR2	76.1	± 0.2	75.9	± 0.2	76.0	± 0.2	75.8	± 0.2	75.9	± 0.2
VIRUSES
Adenovirus C2	Adeno2	89.0	± 0.2	88.8	± 0.1	88.8	± 0.3	88.7	± 0.2	88.8	± 0.2
(ATCC VR-846)	Adeno6	89.6	± 0.2	89.4	± 0.2	89.4	± 0.3	89.2	± 0.2	89.4	± 0.3
Coronavirus OC43
(ATCC VR-759)	CoV-OC43-2	80.7	± 0.2	80.6	± 0.1	80.6	± 0.3	80.5	± 0.2	80.6	± 0.2
Human Metapneumovirus
(Zeptometrix 0810161CF)	hMPV	78.2	± 0.3	78.0	± 0.2	78.0	± 0.3	77.8	± 0.2	78.0	± 0.3
Rhinovirus
(Zeptometrix 0810012CFN)	HRV/EV	84.3	± 0.2	84.2	± 0.2	84.3	± 0.3	84.1	± 0.2	84.2	± 0.2
Influenza A H3N2	FluA-pan1	84.2	± 0.2	84.0	± 0.1	84.0	± 0.3	83.8	± 0.2	84.0	± 0.2
(ATCC VR-810)	FluA-pan2	78.9	± 0.2	78.9	± 0.1	78.9	± 0.2	78.8	± 0.2	78.9	± 0.2
	FluA-H3	82.1	± 0.2	81.9	± 0.2	82.0	± 0.3	81.9	± 0.2	82.0	± 0.2
Influenza B
(Zeptometrix 0810255CF)	FluB	80.4	± 0.3	80.3	± 0.2	80.4	± 0.2	80.2	± 0.2	80.3	± 0.2
Parainfluenza virus 2
(Zeptometrix 0810015CF)	PIV2	83.2	± 0.2	83.1	± 0.2	83.2	± 0.2	83.0	± 0.2	83.1	± 0.2
Parainfluenza virus 4
(Zeptometrix 0810060CF)	PIV4	77.1	± 0.2	77.0	± 0.3	77.2	± 0.3	77.0	± 0.2	77.1	± 0.3
Respiratory Syncytial Virus
(Zeptometrix 0810040ACF)	RSV	81.2	± 0.2	81.1	± 0.2	81.1	± 0.2	81.0	± 0.2	81.1	± 0.2
BACTERIA
Bordetella parapertussis
(Zeptometrix 0801461)	IS 1001	87.7	± 0.2	87.6	± 0.2	87.6	± 0.3	87.5	± 0.2	87.6	± 0.2
Bordetella pertussis
(Zeptometrix 0801459)	ptxP	88.6	± 0.2	88.5	± 0.2	88.5	± 0.3	88.2	± 0.2	88.4	± 0.3
Chlamydia pneumoniae
(ATCC VR-2282)	Cpne	79.6	± 0.3	79.5	± 0.2	79.5	± 0.3	79.3	± 0.2	79.5	± 0.3

Table 27. Reproducibility of Tm (°C) For Select FilmArray Torch and FilmArray Torch and FilmArray 2.0 Systems Mean Tm values are calculated from a combination of Tm values obtained at the 3x LoD and 1x LoD concentrations.

4 Calculated from results at the 3× LoD and 1× LoD test concentrations combined.