Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The summary describes a standard RT-PCR assay and system for detecting viral RNA. There is no mention of AI or ML in the device description, intended use, or performance studies. The technology described is based on established molecular diagnostic techniques.

Therapeutic

No.
This device is an in vitro diagnostic (IVD) test intended for the qualitative detection and differentiation of viral RNA to aid in diagnosis, not to provide therapy or treatment.

Diagnostic

Yes

The "Intended Use / Indications for Use" section explicitly states that the assay is "intended as an aid in the diagnosis of influenza and respiratory syncytial virus infections."

SaMD (Software as a Medical Device)

No

The device description clearly states it is an automated in vitro diagnostic test performed on the Cepheid GeneXpert Xpress System, which is a hardware platform that automates sample extraction, purification, amplification, and detection using real-time PCR. It also utilizes single-use disposable cartridges containing reagents. This involves significant hardware components and chemical processes, not just software.

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 assay is "intended for the in vitro qualitative detection and differentiation of influenza B, and respiratory syncytial virus (RSV) viral RNA." The term "in vitro" is a key indicator of an IVD, meaning it is used to test samples outside of the living body.
Device Description: The "Device Description" section also clearly states, "The Xpert Flu+RSV Xpress Assay is an automated in vitro diagnostic test for qualitative detection and differentiation of influenza A. influenza B, and respiratory syncytial virus (RSV)."
Purpose: The assay is used to analyze biological specimens (nasopharyngeal swabs) to provide information that aids in the diagnosis of influenza and RSV infections. This is the fundamental purpose of an IVD.
Regulatory Context: The mention of a "Predicate Device(s)" with a 510(k) number (K142045) indicates that this device is subject to FDA regulation for medical devices, specifically IVDs.

Therefore, based on the provided text, the Cepheid Xpert Flu-RSV Xpress Assay is definitively an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The Cepheid Xpert Flu-RSV Xpress Assay, performed on the GeneXpert Xpress System, is an automated, multiplex realtime, reverse transcriptase polymerase chain reaction (RT-PCR) assay intended for the in vitro qualitative detection and differentiation of influenza B, and respiratory syncytial virus (RSV) viral RNA. The Xpert Flu+RSV Xpress Assay uses nasopharyngeal swab specimens collected from patients with signs and symptoms of respiratory infection. The Xpert Flu+RSV Xpress Assay is intended as an aid in the diagnosis of influenza and respiratory syncytial virus infections in conjunction with clinical and epidemiological risk factors.

Negative results do not preclude influenza virus or respiratory syncytial virus infection and should not be used as the sole basis for treatment or other patient management decisions.

Performance characteristics for influenza A were established during the 2014-2015 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 department for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.

The Xpert® Nasopharyngeal Sample Collection Kit is designed to collect, preserve, and transport nasopharyngeal swab specimens and to preserve and transport nasal aspirate/wash specimens containing viruses from patients with signs and symptoms of respiratory infection prior to analysis with the Xpert Flu Assay or the Xpert Flu/RSV XC Assay. The Xpert® Nasopharyngeal Sample Collection Kit is designed to collect, preserve, and transport nasopharyngeal swab specimens containing viruses from patients with signs and symptoms of respiratory infection prior to analysis with the Xpert Flu+RSV Xpress Assay.

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

OCC, OOI, JSM

Device Description

The Xpert Flu+RSV Xpress Assay is an automated in vitro diagnostic test for qualitative detection and differentiation of influenza A. influenza B, and respiratory syncytial virus (RSV). The assay is performed on the Cepheid GeneXpert Xpress System (GeneXpert Dx System, GX-I). The GeneXpert Xpress System platform automates and integrates sample extraction, purification, amplification, and detection of the target sequence in simple or complex samples using real-time PCR and reverse transcriptase PCR (RT-PCR) assays. The systems require the use of single-use disposable cartridges (the Xpert Flu+RSV Xpress cartridges) that hold the PCR reagents and host the PCR process. Because the cartridges are self-contained and specimens never come into contact with working parts of the instrument modules, cross-contamination between samples is minimized.

The Xpert Flu+RSV Xpress Assay includes reagents for the detection and differentiation of influenza A, influenza B, and RSV viral RNA directly from nasopharyngeal (NP) swab specimens collected from patients with signs and symptoms of respiratory infection. A Sample Processing Control (SPC) and a Probe Check Control (PCC) are also included in the cartridge. The SPC is present to control for adequate processing of the target viruses and to monitor the presence of inhibitors in the PCR reaction. The Probe Check Control (PCC) verifies reagent rehydration, PCR tube filling in the cartridge, probe integrity, and dye stability.

The single-use, multi-chambered fluidic cartridges are designed to complete sample preparation and real-time RT-PCR for detection and differentiation of influenza B and RSV viral RNA in approximately 60 minutes. The GeneXpert Xpress System, comprised of the GeneXpert Dx System GX-I, has one module that is capable of performing separate sample preparation and real-time PCR and RT-PCR tests. Each module contains a syringe drive for dispensing fluids (i.e., the syringe drive activates the plunger that works in concert with the rotary valve in the cartridge to move fluids between chambers), an ultrasonic horn for lysing cells or spores, and a proprietary I-CORE® thermocycler for performing realtime PCR and RT-PCR and detection.

Specimens are collected following the instructions for collecting NP swab specimens provided in Xpert Flu+RSV Xpress Assay package insert for influenza and RSV testing. The Cepheid Xpert Nasopharyngeal Sample Collection Kit (Cepheid catalog #SWAB/B-100) is required but not provided for use with the assay. The NP swab specimen is placed in the Xpert viral transport medium and sent to the GeneXpert® Xpress testing area for processing. When stored in the transport medium, the NP swab specimen is stable for up to 24 hours at 2-30 °C or up to seven days at 2-8 °C. When ready to test the specimen, the user briefly mixes the specimen by inverting the tube five times, transfers the eluted material to the sample chamber in the top of the disposable fluidic cartridge. The user initiates a test from the system user interface and places the cartridge into the GeneXpert Xpress instrument platform, which performs hands-off real-time, multiplex polymerase chain reaction (PCR) for detection of RNA. The results are automatically generated at the end of the process in a report that can be viewed and printed.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

nasopharyngeal

Indicated Patient Age Range

Not Found

Intended User / Care Setting

Untrained operators with no clinical lab experience in a CLIA-waiver environment.

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

Description of the test set, sample size, data source, and annotation protocol:
A total of 2435 NP swab specimens were tested for influenza A, influenza B and RSV by the Xpert Flu+RSV Xpress Assay and the comparator assay. Of the 2435 NP swab specimens, 2176 were fresh, prospectively collected and 259 were pre-selected frozen, archived specimens.
The Xpert Flu+RSV Xpress Assay performance was compared to a FDA-cleared molecular comparator assay. Xpert Flu+RSV Xpress Assay results from NP swab specimens were compared to the molecular comparator assay result from the same swab specimens. Bidirectional sequencing was performed on specimens where the Xpert Flu-RSV Xpress Assav and the comparator assay were discrepant, and is provided for informational purposes only.

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

Clinical Comparison Study:
Performance characteristics of the Xpert Flu+RSV Xpress Assay were evaluated at 12 institutions in the U.S. during the 2014-2015 influenza season. Due to the low prevalence of Flu B and RSV during the sampling time frame of this study, supplementation with preselected archived NP swab specimens known to be positive for Flu B or RSV was required to meet the sample size for these targets.
Subjects included individuals with signs or symptoms of respiratory infection and whose routine care called for collection of NP swab specimens for influenza and/or RSV testing. For eligible subjects, NP swab specimens were obtained for testing with the Xpert Flu+RSV Xpress Assay and the comparator assay, and patient management continued at the site per their standard practice.
On fresh, prospectively collected NP swab specimens (n=2176), the Xpert Flu+RSV Xpress Assay demonstrated a positive percent agreement (PPA) and negative percent agreement (NPA) for detection of influenza A of 100% and 94.8%, respectively, relative to the comparator assay. The Xpert Flu+RSV Xpress Assay PPA and NPA for influenza B were 100% and 99.5%, respectively. The Xpert Flu+RSV Xpress Assay PPA and NPA for RSV were 96.9% and 99.6%, respectively.
On pre-selected frozen, archived NP swab specimens (n=259), the Xpert Flu+RSV Xpress Assay demonstrated an NPA for detection of influenza A of 98.5% relative to the comparator assay. The Xpert Flu+RSV Xpress Assay PPA and NPA for influenza B were 100% and 98.7%, respectively. The Xpert Flu+RSV Xpress Assay PPA and NPA for RSV were 97.6% and 99.5%, respectively.
Of the Xpert Flu+RSV Xpress Assay runs performed with eligible specimens, 95.0% (2335/2459) of these specimens were successful on the first attempt. The initial invalid rate was 5.0% (95% CI 4.2-6.0%). One-hundred twenty-four gave invalid results on the first attempt (121 NO RESULT-REPEAT TEST and 3 INSTRUMENT ERROR). One-hundred eighteen of the 124 specimens were retested, of which 107 yielded valid results after a single retest. There were 17 NP swab specimens with invalid results upon retest which were excluded in the analyses.

Reproducibility Study:
A panel of 10 specimens with varying concentrations of influenza A. influenza B, and RSV was tested on ten different days by three different operators, at each of three sites (10 specimens x 1 time/day x 10 days x 3 operators x 3 sites). One lot of Xpert Flu-RSV Xpress Assay cartridges was used at each of the 3 testing sites. The Xpert Flu+RSV Xpress Assay was performed according to the Xpert Flu+RSV Xpress Assay procedure.
Results are summarized in Table 5-13 and Table 5-14.

Negative samples: 100% total agreement.
Flu A-High Neg: 29.5% total agreement.
Flu A-Low Pos: 98.9% total agreement.
Flu A-Mod Pos: 100% total agreement.
Flu B-High Neg: 37.1% total agreement.
Flu B-Low Pos: 98.9% total agreement.
Flu B-Mod Pos: 100% total agreement.
RSV-High Neg: 54.5% total agreement.
RSV-Low Pos: 97.8% total agreement.
RSV-Mod Pos: 100% total agreement.

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

On fresh, prospectively collected NP swab specimens:

Flu A: PPA 100% (98.5-100), NPA 94.8% (93.7-95.7)
Flu B: PPA 100% (94.3-100), NPA 99.5% (99.1-99.8)
RSV: PPA 96.9% (92.3-99.1), NPA 99.6% (99.2-99.8)

On pre-selected frozen, archived NP swab specimens:

Flu A: NPA 98.5% (96.1-99.6)
Flu B: PPA 100% (96.4-100), NPA 98.7% (95.5-99.8)
RSV: PPA 97.6% (87.1-99.9), NPA 99.5% (97.5-100)

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.

K142045, K042970

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

0

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 stylized eagle with three heads facing to the right. The eagle is surrounded by a circular border with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" written around it.

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

Cepheid Scott Campbell, PhD, MBA Executive Director, Clinical Affairs 904 Caribbean Drive Sunnyvale, CA 94089

December 3, 2015

Re: K151226

Trade/Device Name: Xpert® Flu+RSV Xpress, Xpert® Nasopharyngeal Sample Collection Kit, GeneXpert Xpress System (GX-I) Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory viral panel multiplex nucleic acid assay Regulatory Class: II Product Code: OCC, OOI, JSM Dated: May 5, 2015 Received: May 8, 2015

Dear Dr. Campbell:

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 (OS) 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/ResourcesforYou/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,

Uwe Scherf -S

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

2

Indications for Use

510(k) Number (if known) K151226

Device Name Xpert Flu+RSV Xpress

Xpert® Nasopharyngeal Sample Collection Kit

Indications for Use (Describe)

The Cepheid Xpert Flu-RSV Xpress Assay, performed on the GeneXpert Xpress System, is an automated, multiplex realtime, reverse transcriptase polymerase chain reaction (RT-PCR) assay intended for the in vitro qualitative detection and differentiation of influenza B, and respiratory syncytial virus (RSV) viral RNA. The Xpert Flu+RSV Xpress Assay uses nasopharyngeal swab specimens collected from patients with signs and symptoms of respiratory infection. The Xpert Flu+RSV Xpress Assay is intended as an aid in the diagnosis of influenza and respiratory syncytial virus infections in conjunction with clinical and epidemiological risk factors.

Negative results do not preclude influenza virus or respiratory syncytial virus infection and should not be used as the sole basis for treatment or other patient management decisions.

Performance characteristics for influenza A were established during the 2014-2015 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 department for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.

The Xpert® Nasopharyngeal Sample Collection Kit is designed to collect, preserve, and transport nasopharyngeal swab specimens and to preserve and transport nasal aspirate/wash specimens containing viruses from patients with signs and symptoms of respiratory infection prior to analysis with the Xpert Flu Assay or the Xpert Flu/RSV XC Assay. The Xpert® Nasopharyngeal Sample Collection Kit is designed to collect, preserve, and transport nasopharyngeal swab specimens containing viruses from patients with signs and symptoms of respiratory infection prior to analysis with the Xpert Flu+RSV Xpress Assay.

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)

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

3

5.0 510(k) Summary

As required by 21 CFR Section 807.92(c).

| Submitted by: | Cepheid
904 Caribbean Drive
Sunnyvale, CA 90489
Phone number: (847) 228-3299
Fax number: (847) 593-0233 |
|--------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Contact: | Scott A. Campbell, PhD, MBA |
| Date of Preparation: | May 5, 2015 |
| Device: | |
| Trade name: | Xpert® Flu+RSV Xpress |
| Common name: | Xpert Flu+RSV Xpress Assay |
| Type of Test: | Automated, multiplex real-time reverse transcription-
polymerase chain reaction (RT-PCR) assay intended for the in vitro qualitative detection and differentiation of influenza A,
influenza B, and respiratory syncytial virus. |
| Regulation number/
Classification name/ | 866.3980/Respiratory viral panel multiplex nucleic acid assay/
866.2570/Instrumentation for clinical multiplex test systems
866.2390/Transport culture medium |
| Product code(s): | OCC, OOI, JSM |
| Classification | Class II |
| Advisory Panel | Microbiology (83) |
| Prescription Use | Yes |
| Predicate Devices
Name(s): | 1) For the detection and differentiation of influenza A,
influenza B, and RSV viral RNA in nasopharyngeal swab
specimens:
Cepheid Xpert Flu/RSV XC [510(k) #K142045]; and,

For the Sample Collection Kits: |

Cepheid Xpert Nasopharyngeal Sample Collection Kit [510(k) #K142045]

4

Device Description:

The Xpert Flu+RSV Xpress Assay is an automated in vitro diagnostic test for qualitative detection and differentiation of influenza A. influenza B, and respiratory syncytial virus (RSV). The assay is performed on the Cepheid GeneXpert Xpress System (GeneXpert Dx System, GX-I). The GeneXpert Xpress System platform automates and integrates sample extraction, purification, amplification, and detection of the target sequence in simple or complex samples using real-time PCR and reverse transcriptase PCR (RT-PCR) assays. The systems require the use of single-use disposable cartridges (the Xpert Flu+RSV Xpress cartridges) that hold the PCR reagents and host the PCR process. Because the cartridges are self-contained and specimens never come into contact with working parts of the instrument modules, cross-contamination between samples is minimized.

The Xpert Flu+RSV Xpress Assay includes reagents for the detection and differentiation of influenza A, influenza B, and RSV viral RNA directly from nasopharyngeal (NP) swab specimens collected from patients with signs and symptoms of respiratory infection. A Sample Processing Control (SPC) and a Probe Check Control (PCC) are also included in the cartridge. The SPC is present to control for adequate processing of the target viruses and to monitor the presence of inhibitors in the PCR reaction. The Probe Check Control (PCC) verifies reagent rehydration, PCR tube filling in the cartridge, probe integrity, and dye stability.

The single-use, multi-chambered fluidic cartridges are designed to complete sample preparation and real-time RT-PCR for detection and differentiation of influenza B and RSV viral RNA in approximately 60 minutes. The GeneXpert Xpress System, comprised of the GeneXpert Dx System GX-I, has one module that is capable of performing separate sample preparation and real-time PCR and RT-PCR tests. Each module contains a syringe drive for dispensing fluids (i.e., the syringe drive activates the plunger that works in concert with the rotary valve in the cartridge to move fluids between chambers), an ultrasonic horn for lysing cells or spores, and a proprietary I-CORE® thermocycler for performing realtime PCR and RT-PCR and detection.

Specimens are collected following the instructions for collecting NP swab specimens provided in Xpert Flu+RSV Xpress Assay package insert for influenza and RSV testing. The Cepheid Xpert Nasopharyngeal Sample Collection Kit (Cepheid catalog #SWAB/B-100) is required but not provided for use with the assay. The NP swab specimen is placed in the Xpert viral transport medium and sent to the GeneXpert® Xpress testing area for processing. When stored in the transport medium, the NP swab specimen is stable for up to 24 hours at 2-30 °C or up to seven days at 2-8 °C. When ready to test the specimen, the user briefly mixes the specimen by inverting the tube five times, transfers the eluted material to the sample chamber in the top of the disposable fluidic cartridge. The user initiates a test from the system user interface and places the cartridge into the GeneXpert Xpress instrument platform, which performs hands-off real-time, multiplex polymerase chain reaction (PCR) for detection of RNA. The results are automatically generated at the end of the process in a report that can be viewed and printed.

5

Device Intended Use:

Xpert Flu+RSV Xpress Assay:

The Cepheid Xpert® Flu+RSV Xpress Assay, performed on the GeneXpert® Xpress System, is an automated, multiplex real-time, reverse transcriptase polymerase chain reaction (RT-PCR) assay intended for the in vitro qualitative detection and differentiation of influenza A, influenza B, and respiratory syncytial virus (RSV) viral RNA. The Xpert Flu+RSV Xpress Assay uses nasopharyngeal swab specimens collected from patients with signs and symptoms of respiratory infection. The Xpert Flu+RSV Xpress Assay is intended as an aid in the diagnosis of influenza and respiratory syncytial virus in conjunction with clinical and epidemiological risk factors.

Negative results do not preclude influenza virus or respiratory syncytial virus infection and should not be used as the sole basis for treatment or other patient management decisions.

Performance characteristics for influenza A were established during the 2014-2015 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.

Ancillary Collection Kits' Indications for Use (the expanded indication is shown in bold):

Xpert Nasopharyngeal Sample Collection Kit

The Xpert® Nasopharyngeal Sample Collection Kit is designed to collect, preserve, and transport nasopharyngeal swab specimens and to preserve and transport nasal aspirate/wash specimens containing viruses from patients with signs and symptoms of respiratory infection prior to analysis with the Xpert Flu Assay or the Xpert Flu/RSV XC Assay. The Xpert® Nasopharyngeal Sample Collection Kit is designed to collect, preserve, and transport nasopharyngeal swab specimens containing viruses from patients with signs and symptoms of respiratory infection prior to analysis with the Xpert Flu+RSV Xpress Assay.

Substantial Equivalence:

The Xpert Flu+RSV Xpress Assay is substantially equivalent to the Cepheid Xpert Flu/RSV XC Assay [510(k) #K142045]. The Xpert Flu+RSV Xpress Assay and the Xpert Flu/RSV XC Assay detect influenza A. influenza B. and RSV from NP swab specimens. Both assays utilize the same technology by determining the presence of the target organisms through realtime RT-PCR amplification and fluorogenic target-specific hybridization detection. A multi

6

center clinical study was conducted to determine the performance characteristics of the device with collection of NP swab specimens from each subject relative to a comparator device, the ProFlu+ Assay which is FDA cleared for NP swab specimens. Discordant results between the Xpert Flu+RSV Xpress Assay and the ProFlu+ Assay were analyzed by sequencing using primers different from those used in the Xpert Flu-RSV Xpress Assay. The study results showed the Xpert Flu+RSV Xpress Assay is acceptable for its intended use with inexperienced lab users.

Table 5-1 shows the similarities and differences between the Xpert Flu+RSV Xpress Assay and the predicate assay, Xpert Flu/RSV XC Assay.

Similarities
Item	Device	Predicate Device
	Cepheid Xpert Flu+RSV
Xpress Assay	Cepheid Xpert Flu/RSV XC
Assay
510(k) Number	K151226	K142045
Regulation	866.3980	866.3980
Product Code	OCC, OOI	OCC, OOI
Device Class	Same	II
Technology Principle
of Operation	Same	Multiplex real time RT-PCR
Similarities
Item	Device	Predicate Device
	Cepheid Xpert Flu+RSV
Xpress Assay	Cepheid Xpert Flu/RSV XC
Assay
Intended Use	The Cepheid Xpert Flu+RSV
Xpress Assay, performed on
the GeneXpert Xpress System,
is an automated, multiplex real-
time, reverse transcriptase
polymerase chain reaction (RT-
PCR) assay intended for the in
vitro qualitative detection and
differentiation of influenza A,
influenza B, and respiratory
syncytial virus (RSV) viral
RNA. The Xpert Flu+RSV
Xpress Assay uses
nasopharyngeal swab specimens
collected from patients with
signs and symptoms of
respiratory infection. The Xpert
Flu+RSV Xpress Assay is
intended as an aid in the
diagnosis of influenza and
respiratory syncytial virus in
conjunction with clinical and
epidemiological risk factors.	The Cepheid Xpert Flu/RSV XC
Assay is an automated,
multiplex real-time, reverse
transcriptase polymerase chain
reaction (RT-PCR assay
intended for the in vitro
qualitative detection and
differentiation of influenza A,
influenza B, and respiratory
syncytial virus (RSV) viral
RNA. The Xpert Flu/RSV XC
Assay uses nasopharyngeal
swab and nasal aspirate/wash
specimens collected from
patients with signs and
symptoms of respiratory
infection in conjunction with
clinical and epidemiological risk
factors. The Xpert Flu/RSV XC
Assay is intended as an aid in
the diagnosis of influenza and
respiratory syncytial virus.
	Negative results do not preclude
influenza virus or respiratory
syncytial virus infection and
should not be used as the sole
basis for treatment or other
patient management decisions.

Performance characteristics for
influenza A were established
during the 2014-2015 influenza
season. When other novel
influenza A viruses are
emerging, performance
characteristics may vary. | Negative results do not preclude
influenza virus or respiratory
syncytial virus infection and
should not be used as the sole
basis for treatment or other
patient management decisions.

Performance characteristics for
influenza A were established
during the 2013-2014 influenza
season. When other novel
influenza A viruses are
emerging, performance
characteristics may vary. |
| Similarities | | |
| Item | Device | Predicate Device |
| | Cepheid Xpert Flu+RSV
Xpress Assay | Cepheid Xpert Flu/RSV XC
Assay |
| Intended Use | 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 department
for testing. Viral culture should
not be attempted in these cases
unless a BSL 3+ facility is
available to receive and culture
specimens. | 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 department
for testing. Viral culture should
not be attempted in these cases
unless a BSL 3+ facility is
available to receive and culture
specimens. |
| Indication for Use | Same | Patients with signs and
symptoms of respiratory
infection in conjunction with
clinical and epidemiological risk
factors. |
| Assay Targets | Same | Influenza A Virus, Influenza B
Virus, and RSV viral RNA |
| Specimen Types | Nasopharyngeal (NP) swab
specimens | Nasopharyngeal (NP) swab
specimens and
Nasal aspirate/wash (NA/W)
specimens |
| Nucleic Acid
Extraction | Yes | Yes |
| Extraction Methods | Sample preparation integrated in
GeneXpert Cartridge and
GeneXpert Xpress System | Sample preparation integrated in
GeneXpert Cartridge and
GeneXpert Instrument System |
| Assay Results | Same | Qualitative |
| Instrument System | Cepheid GeneXpert Xpress
System (instrument model
GX-I); same Cepheid I-core
technology | Cepheid GeneXpert Instrument
Systems (various instrument
models including instrument
model GX-I); Cepheid I-core
technology |
| Similarities | | |
| Item | Device | Predicate Device |
| | Cepheid Xpert Flu+RSV
Xpress Assay | Cepheid Xpert Flu/RSV XC
Assay |
| Assay Controls | Same | Encapsulated (armored) RNA
pseudovirus as a sample
processing control.
Available but not provided are
inactivated virus controls for
influenza A/B and RSV as
external positive controls, and
Coxsackie virus as an external
negative control. |
| Time to obtain test
results | Approximately 60 minutes for
sample preparation and real-
time RT-PCR. | Approximately 60 minutes or
less for sample preparation and
real-time RT-PCR. |
| Primers and probes | Same | Primers and probes to detect the
presence of nucleic acid
sequences of influenza A,
influenza B, and RSV. |

Table 5-1: Comparison of Similarities and Differences of the Xpert Flu+RSV Xpress Assay with the Predicate Device

7

8

9

10

| Primary Differences

(Differences are also Captured in the Similarities Table above)
	New Device	Predicate Device
Item	Cepheid Xpert Flu+RSV
Xpress Assay	Cepheid Xpert Flu/RSV XC
Assay
Instrument System	Cepheid GeneXpert Xpress
System	Cepheid GeneXpert Dx
Systems and GeneXpert
Infinity Systems
Laboratory Users	Untrained operators with no
clinical lab experience in a
CLIA-waiver environment.	Operators with no clinical lab
experience to experienced
clinical laboratory
technologists.
Combinatorial Assay
Selections	No combinatorial assay
selections are available.	Yes, user may select combined
assay with all targets or a Flu
only assay or a RSV only
assay.
Early assay
termination function	No early assay termination
function is available.	Yes, on Flu only or RSV only
assay selections.

The Xpert Flu+RSV Xpress Assay has the same general intended use as the predicate device and has the same technological characteristics as the predicate device. The differences between the Xpert Flu+RSV Xpress Assay and the predicate device do not raise different questions of safety and effectiveness. The clinical study demonstrates that the Xpert Flu+RSV Xpress Assay is acceptable for its intended use with inexperienced laboratory users and is substantially equivalent to the predicate device described above.

Xpert Nasopharyngeal Sample Collection Kit

The predicate device for the ancillary specimen collection kit, the Xpert® Nasopharyngeal Sample Collection Kit, is the Cepheid Nasopharyngeal Sample Collection Kit, [510(k) # K042970]. The similarities and differences are shown in Table 5-2.

11

| Sample Collection Kit with the Predicate Device

Similarities
	Device	Predicate
Item	Xpert Nasopharyngeal Sample
Collection Kit	Xpert Nasopharyngeal Sample
Collection Kit
Intended Use
(Similarities)	For collection, preservation
and transport of
nasopharyngeal swab
specimens and to preserve and
transport nasal aspirate/wash
specimens containing viruses
from patients with signs and
symptoms of respiratory
infection prior to analysis with
the Xpert Flu Assay and the
Xpert Flu/RSV XC Assay.
For collection, preservation and
transport of nasopharyngeal
swab specimens containing
viruses from patients with signs
and symptoms of respiratory
infection prior to analysis with
the Xpert Flu+RSV Xpress
Assay.	For collection, preservation
and transport of
nasopharyngeal swab
specimens and to preserve and
transport nasal aspirate/wash
specimens containing viruses
from patients with signs and
symptoms of respiratory
infection prior to analysis with
the Xpert Flu Assay and the
Xpert Flu/RSV XC Assay.
Single-use Device	Yes	Yes
Medium Formulation	Same	Hank's Balanced Salt Solution
Bovine Serum Albumin
L-cysteine
Gelatin
Sucrose
L-glutamic acid
HEPES buffer
Vancomycin
Amphotericin B
Colistin
Phenol red
pH	Same	$7.3 \pm 0.2$

Table 5-2: Comparison of Similarities and Differences of the Xpert Nasopharyngeal
Sample Collection Kit with the Predicate Device

12

Similarities
	Device	Predicate
Item	Xpert Nasopharyngeal Sample
Collection Kit	Xpert Nasopharyngeal Sample
Collection Kit
Storage Temperature	Same	2 - 25°C (refrigerated
and room temperature)
Volume	Same	3 ml
Glass Beads	Same	3 x 3 mm
Container	Same	Plastic (medical-grade
polypropylene)
Product Configuration	Same	Medium Tube in Kit with
individually-wrapped sterile
swab.
Differences
Item	Device	Predicate
Intended Use
(differences)	For collection, preservation and
transport of nasopharyngeal
swab specimens containing
viruses from patients with signs
and symptoms of respiratory
infection prior to analysis with
the Xpert Flu+RSV Xpress
Assay.	For collection, preservation and
transport of nasopharyngeal
swab specimens and to preserve
and transport nasal
aspirate/wash specimens
containing viruses from patients
with signs and symptoms of
respiratory infection prior to
analysis with the Xpert Flu
Assay and the Xpert Flu/RSV
XC Assay.

Both devices have the same general intended use and use the same technology to collect, store, and transport clinical specimens, including viruses, to the laboratory for further testing. The differences between the two devices do not raise new or different questions of safety and effectiveness. The multi-center clinical study of the Xpert Flu+RSV Xpress Assay was conducted using Xpert Nasopharyngeal Sample Collection Kit and demonstrated that the assay and its collection kit are acceptable for their intended use with inexperienced lab users and substantially equivalent to the predicate device.

13

Non-Clinical Studies:

Analytical Sensitivity (Limit of Detection)

Studies were performed to determine the analytical limit of detection (LoD) of the Xpert Flu+RSV Xpress Assay with two lots of reagents across three testing days. The higher LoD observed per strain and per lot was selected for verification. Verification of the estimated LoD claim was performed on one reagent lot across a minimum of three testing days. LoD was established using two influenza A H3N2 strains, two influenza A 2009 H1N1 strains, two influenza B strains, two respiratory syncytial virus A (RSV A) strains, two respiratory syncytial virus B (RSV B) strains, and one influenza A H7N9 strain diluted into a negative pooled clinical matrix. The LoD is defined as the lowest concentration (tissue culture infective dose, TCID50/mL) per sample that can be reproducibly distinguished from negative samples with 95% confidence or the lowest concentration at which 19 of 20 replicates were positive. Each strain was tested in replicates of 20 per concentration of virus.

The LoD was determined empirically as the first concentration that had 19/20 or 20/20 positive results. The LoD point estimates for each strain tested are summarized in Tables 5-3 to 5-8.

| Strain ID | Confirmed LoD
(TCID50/mL)
(at least 19/20 positive) |
|-------------------------------|-----------------------------------------------------------|
| Influenza A/California/7/2009 | 0.3 (20/20) |
| Influenza A/Florida/27/2011 | 16.0 (20/20) |

Table 5-3: Confirmed LoD (TCID50/mL): Influenza A 2009 H1N1

Table 5-4: Confirmed LoD (TCID50/mL): Influenza A H3N2

| Strain ID | Confirmed LoD
(TCID50/mL)
(at least 19/20 positive) |
|-------------------------------|-----------------------------------------------------------|
| Influenza A/Perth/16/2009 | 0.3 (20/20) |
| Influenza A/Victoria/361/2011 | 0.8 (20/20) |

Table 5.5: Confirmed LoD (TCID50/mL): Influenza B

| Strain ID | Confirmed LoD
(TCID50/mL)
(at least 19/20 positive) |
|----------------------------------|-----------------------------------------------------------|
| Influenza B/Massachusetts/2/2012 | 0.5 (20/20) |
| Influenza B/Wisconsin/01/2011 | 0.6 (20/20) |

14

| Strain ID | Confirmed LoD
(TCID50/mL)
(at least 19/20 positive) |
|----------------------|-----------------------------------------------------------|
| RSV A/2/Australia/61 | 1.2 (20/20) |
| RSV A/Long/MD/56 | 1.0 (20/20) |

Table 5-6: Confirmed LoD (TCID50/mL): Respiratory Syncytial Virus A

Table 5-7: Confirmed LoD (TCID50/mL): Respiratory Syncytial Virus B

| Strain ID | Confirmed LoD
(TCID50/mL)
(at least 19/20 positive) |
|-----------------------------|-----------------------------------------------------------|
| RSV B/Washington/18537/62 | 1.8 (20/20) |
| RSV B/9320/Massachusetts/77 | 2.0 (20/20) |

Table 5-8: Confirmed LoD (TCID50/mL): Influenza A H7N9

| Strain ID | Confirmed LoD
(TCID50/mL)
(at least 19/20 positive) |
|--------------------------|-----------------------------------------------------------|
| Influenza A/Anhui/1/2013 | 0.8 (19/20) |

Although this test has been shown to detect the novel avian influenza A (H7N9) cultured material, the performance characteristics of this device with clinical specimens that are positive for the novel avian influenza A (H7N9) virus have not been established. The Xpert Flu+RSV Xpress Assay can distinguish between influenza A and B viruses, but it cannot differentiate influenza A subtypes.

Analytical Specificity (Exclusivity)

The analytical specificity of the Xpert Flu+RSV Xpress Assay was evaluated by testing a panel of 44 cultures consisting of 16 viral, 26 bacterial, and two yeast strains representing common respiratory pathogens or those potentially encountered in the nasopharynx. Three replicates of all bacterial and yeast strains were tested at concentrations of ≥ 100 CFU/mL with the exception of one strain which was tested at 10' CFU/mL (Chlamydia pneumoniae). Three replicates of each virus were tested at concentrations of ≥ 105 TCID50/mL. The analytical specificity was 100%. Results are shown in Table 5-9.

Table 5-9: Analytical Specificity of Xpert Flu+RSV Xpress Assay

15

Organism	Concentration	Influenza A	Influenza B	RSV
No Template Control		NEG	NEG	NEG
Adenovirus Type 1	1.12x107 TCID50/mL	NEG	NEG	NEG
Adenovirus Type 7	1.87x105 TCID50/mL	NEG	NEG	NEG
Human coronavirus OC43	2.85x105 TCID50/mL	NEG	NEG	NEG
Human coronavirus 229E	1x105 TCID50/mL	NEG	NEG	NEG
Cytomegalovirus	7.24x105 TCID50/mL	NEG	NEG	NEG
Echovirus	3.31x107 TCID50/mL	NEG	NEG	NEG
Enterovirus	1x105 TCID50/mL	NEG	NEG	NEG
Epstein Barr Virus	7.16x107 TCID50/mL	NEG	NEG	NEG
HSV	8.9x106 TCID50/mL	NEG	NEG	NEG
Measles	6.3x105 TCID50/mL	NEG	NEG	NEG
Human metapneumovirus	3.8x105 TCID50/mL	NEG	NEG	NEG
Mumps virus	6.31x106 TCID50/mL	NEG	NEG	NEG
Human parainfluenza Type 1	1.15x106 TCID50/mL	NEG	NEG	NEG
Human parainfluenza Type 2	1x105 TCID50/mL	NEG	NEG	NEG
Human parainfluenza Type 3	3.55x107 TCID50/mL	NEG	NEG	NEG
Rhinovirus Type 1A	1.26x105 TCID50/mL	NEG	NEG	NEG
Acinetobacter baumannii	>1x106 CFU/mL	NEGa	NEG	NEG
Burkholderia cepacia	>1x106 CFU/mL	NEG	NEG	NEG
Candida albicans	>1x106 CFU/mL	NEG	NEG	NEG
Candida parapsilosis	>1x106 CFU/mL	NEG	NEG	NEG
Bordetella pertussis	1x108 CFU/mL	NEG	NEG	NEG
Chlamydia pneumoniae	3.16x105 CFU/mL	NEG	NEG	NEG
Organism	Concentration	Influenza A	Influenza B	RSV
Citrobacter freundii	>1x106 CFU/mL	NEG	NEG	NEG
Corynebacterium sp.	>1x106 CFU/mL	NEG	NEG	NEG
Escherichia coli	>1x106 CFU/mL	NEG	NEG	NEG
Enterococcus faecalis	>1x106 CFU/mL	NEG	NEG	NEG
Haemophilus influenzae	1x106 CFU/mL	NEG	NEG	NEG
Lactobacillus reuter	1x106 CFU/mL	NEG	NEG	NEG
Legionella spp.	1x108 CFU/mL	NEG	NEG	NEG
Moraxella catarrhalis	>1x106 CFU/mL	NEG	NEG	NEG
Mycobacterium tuberculosis
(avirulent)	1.15x106 CFU/mL	NEG	NEG	NEG
Mycoplasma pneumoniae	1x107 CFU/mL	NEG	NEG	NEG
Neisseria meningitidis	>1x106 CFU/mL	NEG	NEG	NEG
Neisseria mucosa	>1x106 CFU/mL	NEG	NEG	NEG
Propionibacterium acnes	>1x106 CFU/mL	NEG	NEG	NEG
Pseudomonas aeruginosa	>1x106 CFU/mL	NEG	NEG	NEG
Staphylococcus aureus	>1x106 CFU/mL	NEG	NEG	NEG
Staphylococcus epidermidis	>1x106 CFU/mL	NEG	NEG	NEG
Staphylococcus haemolyticus	>1x106 CFU/mL	NEG	NEG	NEG
Streptococcus agalactiae	>1x106 CFU/mL	NEG	NEG	NEG
Streptococcus pneumoniae	>1x106 CFU/mL	NEG	NEG	NEG
Streptococcus pyogenes	>1x106 CFU/mL	NEG	NEG	NEG
Streptococcus salivarius	>1x106 CFU/mL	NEG	NEG	NEG
Streptococcus sanguinis	>1x106 CFU/mL	NEG	NEG	NEG

16

Analytical Reactivity (Inclusivity)

The analytical reactivity of the Xpert Flu+RSV Xpress Assay was evaluated against multiple strains of influenza A H1N1 (seasonal pre-2009), influenza A H1N1 (pandemic 2009), influenza A H3N2 (seasonal), avian influenza A (H5N1, H5N2, H6N2, H7N2, H7N3, H2N2, H7N9, and H9N2), influenza B (representing strains from both Victoria and

17

Yamagata lineages), and respiratory syncytial virus subgroups A and B (RSV A and RSV B) at levels near the analytical LoD. A total of 64 strains including 54 influenza viruses and 10 RSV strains were tested in this study with the Xpert Flu+RSV Xpress Assay.

Three replicates were tested for each strain. Results are shown in Table 5-10.

Virus	Strain	Concentration	Result
			Flu A	Flu B	RSV
No Template Control			NEG	NEG	NEG
Influenza A
H1N1 (pre-2009)	A/swine/Iowa/15/30	32.0 TCID50/mL	POS	NEG	NEG
	A/WS/33	32.0 TCID50/mL	POS	NEG	NEG
	A/PR/8/34	32.0 TCID50/mL	POS	NEG	NEG
	A/Mal/302/54	32.0 TCID50/mL	POS	NEG	NEG
	A/Denver/1/57	32.0 TCID50/mL	POS	NEG	NEG
	A/New Jersey/8/76	32.0 TCID50/mL	POS	NEG	NEG
	A/New Caledonia/20/1999	32.0 TCID50/mL	POS	NEG	NEG
	A/New York/55/2004	32.0 TCID50/mL	POS	NEG	NEG
	A/Soloman Islands/3/2006	32.0 TCID50/mL	POS	NEG	NEG
	A/Taiwan/42/06	32.0 TCID50/mL	POS	NEG	NEG
	A/Brisbane/59/2007	32.0 TCID50/mL	POS	NEG	NEG
	A/California/7/2009	32.0 TCID50/mL	POS	NEG	NEG
Influenza A
H1N1
(pdm2009)	A/swine/NY/02/2009	32.0 TCID50/mL	POS	NEG	NEG
	A/Florida/27/2011	32.0 TCID50/mL	POS	NEG	NEG
	A/Colorado/14/2012	32.0 TCID50/mL	POS	NEG	NEG
	A/Washington/24/2012	80.0ª TCID50/mL	POS	NEG	NEG

	A/Aichi/2/68	1.6 TCID50/mL	POS	NEG	NEG
	A/HongKong/8/68	1.6 TCID50/mL	POS	NEG	NEG
	A/Port Chalmers/1/73	1.6 TCID50/mL	POS	NEG	NEG
	A/Hawaii/15/2001	1.6 TCID50/mL	POS	NEG	NEG
Influenza A
H3N2
(Seasonal)	A/Wisconsin/67/05	1.6 TCID50/mL	POS	NEG	NEG
	A/Brisbane/10/2007	1.6 TCID50/mL	POS	NEG	NEG
	A/Perth/16/2009	1.6 TCID50/mL	POS	NEG	NEG
	A/Minnesota/11/2010 (H3N2)v	1.6 TCID50/mL	POS	NEG	NEG
	A/Indiana/08/2011 (H3N2)v	1.6 TCID50/mL	POS	NEG	NEG
	A/Victoria/361/2011	1.6 TCID50/mL	POS	NEG	NEG
	A/Texas/50/2012	1.6 TCID50/mL	POS	NEG	NEG
Virus	Strain	Concentration	Result
			Flu A	Flu B	RSV
	A/duck/Hunan/795/2002 (H5N1)	≤ 1pg/μLb	POS	NEG	NEG
	A/chicken/Hubei/327/2004 (H5N1)	≤ 1pg/μLb	POS	NEG	NEG
	A/Anhui/01/2005 (H5N1)	≤ 1pg/μLb	POS	NEG	NEG
	A/Japanese white
eye/HongKong/1038/2006 (H5N1)	≤ 1pg/μLb	POS	NEG	NEG
	A/mallard/WI/34/75 (H5N2)	≤ 1pg/μLb	POS	NEG	NEG
	A/chicken/CA431/00 (H6N2)	≤ 1pg/μLb	POS	NEG	NEG
Avian
influenza A	A/duck/LTC-10-82743/1943 (H7N2)	≤ 1pg/μLb	POS	NEG	NEG
	A/chicken/NJ/15086-3/94 (H7N3)	≤ 1pg/μLb	POS	NEG	NEG
	A/Anhui/1/2013 (H7N9)	N/Ac	POS	NEG	NEG
	A/Shanghai/1/2013 (H7N9)	N/Ac	POS	NEG	NEG
	A/chicken/Korea/38349-p96323/ 1996
(H9N2)	≤ 1pg/μLb	POS	NEG	NEG
	A/mallard/NY/6750/78 (H2N2)	≤ 1pg/μLb	POS	NEG	NEG
	B/Lee/40	1.2 TCID50/mL	NEG	POS	NEG
	B/Allen/45	1.2 TCID50/mL	NEG	POS	NEG
	B/GL/1739/54	1.2 TCID50/mL	NEG	POS	NEG
	B/Maryland/1/59	1.2 TCID50/mL	NEG	POS	NEG
	B/Panama/45/90d	3.0 TCID50/mLe	NEG	POS	NEG
	B/Florida/07/2004f	1.2 TCID50/mL	NEG	POS	NEG
	B/Florida/02/06d	1.2 TCID50/mL	NEG	POS	NEG
Influenza B	B/Florida/04/06	1.2 TCID50/mL	NEG	POS	NEG
	B/Wisconsin/01/2011d	1.2 TCID50/mL	NEG	POS	NEG
	B/Massachusetts/2/2012f	1.2 TCID50/mL	NEG	POS	NEG
	B/Hong Kong/5/72	1.2 TCID50/mL	NEG	POS	NEG
	B/Wisconsin/01/2010f	1.2 TCID50/mL	NEG	POS	NEG
	B/Malaysia/2506/04d	1.2 TCID50/mL	NEG	POS	NEG
	B/Taiwan/2/62	1.2 TCID50/mL	NEG	POS	NEG
	B/Brisbane/60/2008d	1.2 TCID50/mL	NEG	POS	NEG
RSV A	RSV-A/Long/MD/56	2.4 TCID50/mL	NEG	NEG	POS
Virus	Strain	Concentration	Result
			Flu A	Flu B	RSV
	RSV-A/2/Australia/61	2.4 TCID50/mL	NEG	NEG	POS
	RSV-A/NY (Clinical unknown)	2.4 TCID50/mL	NEG	NEG	POS
	RSV-A/WI/629-8-2/2007	2.4 TCID50/mL	NEG	NEG	POS
	RSV-A/WI/629-11-1/2008	2.4 TCID50/mL	NEG	NEG	POS
RSV B	RSV-B/Wash/18537/62	4.0 TCID50/mL	NEG	NEG	POS
	RSV-B/9320/MA/77	4.0 TCID50/mL	NEG	NEG	POS
	RSV-B/WV14617/85	4.0 TCID50/mL	NEG	NEG	POS
	RSV-B/CH93(18)-18	20.0 TCID50/mLg	NEG	NEG	POS
	RSV-B/WI/629-5B/0607	4.0 TCID50/mL	NEG	NEG	POS

Table 5-10: Analytical Reactivity (Inclusivity) of Xpert Flu+RSV Xpress Assay

18

19

"Influenza A/Washington/24/2012 was tested at 5X LoD (80.0 TCID56/mL) to obtain 3 of 3 Flu A POSITIVE result calls.

"Purified viral RNA in simulated background matrix was used for avian influenza A viruses due to biosafety regulations.

Shactivated avian influenza A (H7N9) viruses without viral titer was diluted 100,000 fold in simulated background matrix and tested due to biosafety regulations.

4Known Victoria lineage.

SInfluenza B/Panama/45/90 was tested at 5X LoD (3.0 TCID30mL) to obtain 3/3 Flu B POSITIVE result calls. 4Known Yamagata lineage.

6RSV-B/CH93(18)-18 was tested at 10X LoD (20.0 TCID50/mL) to obtain 3/3 RSV POSITIVE result calls.

20

Potentially Interfering Substances

In a non-clinical study, potentially interfering substances that may be present in the nasopharynx were evaluated directly relative to the performance of the Xpert Flu+RSV Xpress Assay. Potentially interfering substances in the nasopharynx may include, but are not limited to: blood, nasal secretions or mucus, and nasal and throat medications used to relieve congestion, nasal dryness, irritation, or asthma and allergy symptoms, as well as antibiotics and antivirals. Negative samples (n = 8) were tested per each substance to determine the effect on the performance of the sample processing control (SPC). Positive samples (n = 8) were tested per substance with six influenza (four influenza A and two influenza B) and four RSV (two RSV A and two RSV B) strains spiked at 2X the analytical LoD determined for each strain. All results were compared to positive and negative Universal Transport Medium (UTM) controls.

These evaluated substances are listed in Table 5.11 with active ingredients and concentrations tested shown. There was no assay interference in the presence of the substances at the concentrations tested in this study. All positive and negative replicates were correctly identified using the Xpert Flu+RSV Xpress Assay.

FluMist vaccine samples were correctly reported as Flu A POSITIVE; FLU B POSITIVE; RSV NEGATIVE as expected. Samples containing FluMist may cause false positive results. This is addressed in the device labeling Section 17. Limitations.

Substance/Class	Description/Active Ingredient	Concentration Tested
Beta-adrenergic
bronchodilator	Albuterol Sulfate	0.83 mg/mL
(equivalent to
1 dose per day)
Blood	Blood (Human)	2% (v/v)
BDTM Universal
Viral Transport
System	Transport Media	100% (v/v)
Remel M4	Transport Media	100% (v/v)
Remel M4RT	Transport Media	100% (v/v)
Remel M5®	Transport Media	100% (v/v)
Throat lozenges,
oral anesthetic
and analgesic	Benzocaine, Menthol	1.7 mg/mL
Mucin	Purified Mucin protein
(Bovine or porcine
submaxillary gland)	2.5% (w/v)
Antibiotic, nasal
ointment	Mupirocin	10 mg/mL
Saline Nasal
Spray	Sodium Chloride (0.65%)	15% (v/v)
Anefrin Nasal	Oxymetazoline, 0.05%	15% (v/v)

Table 5-11. Potentially Interfering Substances in Xpert Flu+RSV Xpress Assay

21

| Substance/Class | Description/Active
Ingredient | Concentration
Tested |
|-------------------------------|--------------------------------------------------------------------------------|-------------------------|
| Spray | | |
| Nasal Drops | Phenylephrine, 0.5% | 15% (v/v) |
| Tamiflu®/Anti-
viral drugs | Zanamivir | 7.5 mg/mL |
| Antibacterial,
systemic | Tobramycin | 4 µg/mL |
| Zicam®/Nasal
Gel | Luffa opperculata,
Galphimia glauca,
Histaminum
hydrochloricum Sulfur | 15% (w/v) |
| FluMist® | Live intranasal influenza
virus vaccine | 6.7% (v/v) |
| Nasal
corticosteroid | Fluticasone Propionate | 5 µg/mL |

Carry-Over Contamination

A study was conducted to demonstrate that single-use, self-contained GeneXpert cartridges prevent carry-over contamination in negative samples run following very high positive samples in the same GeneXpert module. The study consisted of a negative sample processed in the same GeneXpert module immediately following a very high influenza A sample (approximately 106 TCID5(/test) or a very high RSV A sample (approximately 100 TCID50/test). This testing scheme was repeated 20 times on two GeneXpert modules for a total of 82 runs resulting in 40 positive and 42 negative specimens for each virus type. All 40 positive samples were correctly reported as Flu A POSITIVE; Flu B NEGATIVE; RSV NEGATIVE or Flu A NEGATIVE; Flu B NEGATIVE; RSV POSITIVE. All 42 negative samples were correctly reported as Flu A NEGATIVE; Flu B NEGATIVE; RSV NEGATIVE.

Fresh vs. Frozen Sample Equivalency Study

Fresh and frozen specimen equivalency in the Xpert Flu+RSV Xpress Assay was evaluated by testing individual influenza and RSV strains at three different concentrations representing low positives (2X LoD), moderate positives (5X LoD), and high positives (10X LoD) in simulated background matrix. Negative samples consisted of simulated background matrix only. Fresh and frozen specimen equivalency was determined using one seasonal Flu A H3N2 strain (A/Victoria/361/2011), one Flu B strain (B/Wisconsin/01/11), one RSV A strain (RSV A/Long/MD/56), and one RSV B strain (RSV B/9320/MA/77). Replicates of 20 were tested for each specimen type and concentration. All positive and negative specimens were tested fresh, after one freeze-thaw cycle, and after two freeze-thaw cycles.

There was no statistically significant effect in the performance of the Xpert Flu+RSV Xpress Assay between fresh virus dilutions and two sequential freeze thaw cycles for positive and negative samples. All positive and negative replicates were correctly identified using the Xpert Flu+RSV Xpress Assay.

22

Linearity

Not applicable, the Xpert Flu+RSV Xpress Assay is a qualitative assay.

Clinical Studies

Clinical Comparison Study

Performance characteristics of the Xpert Flu+RSV Xpress Assay were evaluated at 12 institutions in the U.S. during the 2014-2015 influenza season. Due to the low prevalence of Flu B and RSV during the sampling time frame of this study, supplementation with preselected archived NP swab specimens known to be positive for Flu B or RSV was required to meet the sample size for these targets.

Subjects included individuals with signs or symptoms of respiratory infection and whose routine care called for collection of NP swab specimens for influenza and/or RSV testing. For eligible subjects, NP swab specimens were obtained for testing with the Xpert Flu+RSV Xpress Assay and the comparator assay, and patient management continued at the site per their standard practice.

The Xpert Flu+RSV Xpress Assay performance was compared to a FDA-cleared molecular comparator assay. Xpert Flu+RSV Xpress Assay results from NP swab specimens were compared to the molecular comparator assay result from the same swab specimens. Bidirectional sequencing was performed on specimens where the Xpert Flu-RSV Xpress Assav and the comparator assay were discrepant, and is provided for informational purposes only.

NP Swab Specimens

A total of 2435 NP swab specimens were tested for influenza A, influenza B and RSV by the Xpert Flu+RSV Xpress Assay and the comparator assay. Of the 2435 NP swab specimens 2176 were fresh, prospectively collected and 259 were pre-selected frozen, archived specimens.

On fresh, prospectively collected NP swab specimens, the Xpert Flu+RSV Xpress Assay demonstrated a positive percent agreement (PPA) and negative percent agreement (NPA) for detection of influenza A of 100% and 94.8%, respectively, relative to the comparator assay (Table 5-12). The Xpert Flu+RSV Xpress Assay PPA and NPA for influenza B were 100% and 99.5%, respectively (Table 5-12). The Xpert Flu+RSV Xpress Assay PPA and NPA for RSV were 96.9% and 99.6%, respectively (Table 5-12).

On pre-selected frozen, archived NP swab specimens, the Xpert Flu+RSV Xpress Assay demonstrated an NPA for detection of influenza A of 98.5% relative to the comparator assay (Table 5-12). The Xpert Flu+RSV Xpress Assay PPA and NPA for influenza B were 100% and 98.7%, respectively (Table 5-12). The Xpert Flu+RSV Xpress Assay PPA and NPA for RSV were 97.6% and 99.5%, respectively (Table 5-12).

| Specimen
Type | Target | n | TP | FP | TN | FN | PPA %
(95 CI) | NPA %
(95 CI) |
|------------------|--------|------|-----|------|------|----|-------------------|---------------------|
| Fresh | Flu A | 2176 | 250 | 101a | 1825 | 0 | 100
(98.5-100) | 94.8
(93.7-95.7) |

Table 5-12: Xpert Flu+RSV Xpress Assay Performance on NP Swab Specimens

23

| | Flu B | 2176 | 63 | 10b | 2103 | 0 | 100
(94.3-100) | 99.5
(99.1-99.8) |
|------------------------|-------|------|-----|-----|------|----|---------------------|---------------------|
| | RSV | 2176 | 125 | 8c | 2039 | 4d | 96.9
(92.3-99.1) | 99.6
(99.2-99.8) |
| Pre-selected
Frozen | Flu A | 259 | 0 | 4e | 255 | 0 | NA | 98.5
(96.1-99.6) |
| | Flu B | 259 | 100 | 2f | 157 | 0 | 100
(96.4-100) | 98.7
(95.5-99.8) |
| | RSV | 259 | 40 | 1g | 217 | 1h | 97.6
(87.1-99.9) | 99.5
(97.5-100) |

a. Testing results by sequencing: 92 of 101 were Flu A positive: 8 of 101 failed to sequence: 1 of 101 insufficient remaining volume for sequencing.

b. Testing results by sequencing: 9 of 10 were Flu B positive; 1 of 10 insufficient remaining volume for sequencing.

c. Testing results by sequencing: 7 of 8 were RSV positive; 1 of 8 was RSV negative.

d. Testing results by sequencing: 3 of 4 were RSV positive; 1 of 4 was RSV negative.

e. Testing results by sequencing: 4 of 4 insufficient remaining volume for sequencing.

f. Testing results by sequencing: 2 of 2 insufficient remaining volume for sequencing.

g. Testing results by sequencing: 1 of 1 insufficient remaining volume for sequencing.

h. Testing results by sequencing: 1 of 1 insufficient remaining volume for sequencing.

Of the Xpert Flu+RSV Xpress Assay runs performed with eligible specimens, 95.0% (2335/2459) of these specimens were successful on the first attempt. The initial invalid rate was 5.0% (95% CI 4.2-6.0%). One-hundred twenty-four gave invalid results on the first attempt (121 NO RESULT-REPEAT TEST and 3 INSTRUMENT ERROR). One-hundred eighteen of the 124 specimens were retested, of which 107 yielded valid results after a single retest. There were 17 NP swab specimens with invalid results upon retest which were excluded in the analyses.

Reproducibility Study

A panel of 10 specimens with varying concentrations of influenza A. influenza B, and RSV was tested on ten different days by three different operators, at each of three sites (10 specimens x 1 time/day x 10 days x 3 operators x 3 sites). One lot of Xpert Flu-RSV Xpress Assay cartridges was used at each of the 3 testing sites. The Xpert Flu+RSV Xpress Assay was performed according to the Xpert Flu+RSV Xpress Assay procedure. Results are summarized in Table 5-13.

| Sample
ID | Site 1 | | | Site 2 | | | Site 3 | | | % Total
Agreement
by Samplea |
|--------------|-----------------|-----------------|-----------------|-----------------|-----------------|----------------|----------------|-----------------|-----------------|------------------------------------|
| | Op 1 | Op 2 | Op 3 | Op 1 | Op 2 | Op 3 | Op 1 | Op 2 | Op 3 | |
| Negative | 100%
(10/10) | 100%
(10/10) | 100%
(10/10) | 100%
(10/10) | 100%
(10/10) | 100%
(9/9)b | 100%
(9/9)b | 100%
(10/10) | 100%
(10/10) | 100%
(88/88)b |

Table 5-13: Summary of Reproducibility Results

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Sample	Site 1			Site 2			Site 3			% Total
Flu A-High Neg	0.0%
(0/10)	20.0%
(2/10)	60.0%
(6/10)	20.0%
(2/10)	10.0%
(1/10)	50.0%
(5/10)	75.0%
(6/8)b	20.0%
(2/10)	20.0%
(2/10)	29.5%
(26/88)b
Flu A-Low Pos	100%
(10/10)	100%
(10/10)	90.0%
(9/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	98.9%
(89/90)
Flu A-Mod Pos	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100% (90/90)
Flu B-High Neg	50.0%
(5/10)	0.0%
(0/10)	50.0%
(5/10)	20.0%
(2/10)	10.0%
(1/10)	55.6%
(5/9)b	80.0%
(8/10)	50.0%
(5/10)	20.0%
(2/10)	37.1%
(33/89)b
Flu B-Low Pos	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	90.0%
(9/10)	100%
(10/10)	100%
(10/10)	98.9%
(89/90)
Flu B-Mod Pos	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100% (90/90)
RSV-High Neg	60.0%
(6/10)	30.0%
(3/10)	66.7%
(6/9)b	50.0%
(5/10)	60.0%
(6/10)	50.0%
(5/10)	90.0%
(9/10)	33.3%
(3/9)b	50.0%
(5/10)	54.5%
(48/88)b
RSV-Low Pos	88.9%
(8/9)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	90.0%
(9/10)	100%
(10/10)	100%
(10/10)	97.8%
(87/89)c
RSV-Mod Pos	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100%
(10/10)	100% (90/90)

a. Percent of samples yielding expected results – negative for Neg and High Neg samples; positive for Low Pos samples.

b. Seven samples (2 Neg, 2 Flu A High Neg, and 2 RSV High Neg) were indeterminate upon initial and retest.

c. One RSV Low Pos sample was inadvertently not tested.

The reproducibility of the Xpert Flu+RSV Xpress Assay was also evaluated in terms of the fluorescence signal expressed in Ct values for each target detected. The mean, standard deviation (SD), and coefficient of variation (CV) between-sites, between-days, and betweenoperators for each panel member are presented in Table 5-14. One replicate was performed per day per operator, therefore, operator and assay (within-run) precision are confounded.

25

| Sample | Assay
Channel
(Analyte) | Na | Mean
Ct | Between-
Site | | Between-
Day | | Between-
Operator +
Within Assay | | Total | |
|---------------------|-------------------------------|----|------------|------------------|------------|-----------------|------------|----------------------------------------|------------|-------|------------|
| | | | | SD | CV
(%)b | SD | CV
(%)b | SD | CV
(%)b | SD | CV
(%)b |
| Neg | SPC | 88 | 31.4 | 0.3 | 0.9 | 0.0 | 0.0 | 1.5 | 4.7 | 1.5 | 4.8 |
| Flu A -
High Neg | Flu A1 | 62 | 36.9 | 0.1 | 0.3 | 0.6 | 1.6 | 1.6 | 4.4 | 1.7 | 4.7 |
| Flu A -
Low Pos | Flu A1 | 89 | 34.5 | 0.2 | 0.7 | 0.0 | 0.0 | 1.6 | 4.5 | 1.6 | 4.5 |
| Flu A -
Mod Pos | Flu A1 | 90 | 32.3 | 0.0 | 0.0 | 0.7 | 2.2 | 1.2 | 3.7 | 1.4 | 4.2 |
| Flu B -
High Neg | Flu B | 56 | 34.6 | 0.0 | 0.0 | 1.4 | 4.0 | 2.1 | 6.0 | 2.5 | 7.2 |
| Flu B -
Low Pos | Flu B | 89 | 32.6 | 0.6 | 2.0 | 0.7 | 2.1 | 1.7 | 5.1 | 1.9 | 5.9 |
| Flu B -
Mod Pos | Flu B | 90 | 30.4 | 0.0 | 0.0 | 0.0 | 0.0 | 1.0 | 3.3 | 1.0 | 3.3 |
| RSV -
High Neg | RSV | 40 | 36.9 | 1.2 | 3.1 | 0.0 | 0.0 | 1.7 | 4.6 | 2.1 | 5.6 |
| RSV -
Low Pos | RSV | 87 | 35.0 | 0.7 | 1.9 | 0.0 | 0.0 | 1.6 | 4.6 | 1.7 | 5.0 |
| RSV -
Mod Pos | RSV | 90 | 32.6 | 0.0 | 0.0 | 0.1 | 0.4 | 1.0 | 3.0 | 1.0 | 3.0 |

Table 5-14: Summary of Reproducibility Data

a. Results with non-zero Ct values out of 90.

b. (%) is contribution of variance component to overall CV.

Conclusions

The results of the nonclinical analytical and clinical performance studies summarized above demonstrate that the Xpert Flu+RSV Xpress Assay performs comparably to the predicate device for its intended use with inexperienced users in a CLIA waived environment and is substantially equivalent to the predicate device.