Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The description focuses on automated real-time PCR and controls, with no mention of AI or ML in the device description, intended use, or performance studies.

Therapeutic

No

Explanation: The device is an in vitro diagnostic test intended to aid in the diagnosis of trichomoniasis by detecting Trichomonas vaginalis genomic DNA. It does not provide any treatment or therapy.

Diagnostic

Yes

Explanation: The "Intended Use / Indications for Use" section explicitly states, "The Xpert TV Assay is intended to aid in the diagnosis of trichomoniasis in symptomatic or asymptomatic individuals." It also refers to the device as a "qualitative in vitro diagnostic test."

SaMD (Software as a Medical Device)

No

The device is an in vitro diagnostic test that utilizes hardware (GeneXpert Instrument Systems, cartridges) and reagents to perform real-time PCR. While software is involved in controlling the instrument and processing results, it is not a standalone software-only medical device.

IVD (In Vitro Diagnostic)

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

The document explicitly states in multiple places that the Cepheid Xpert TV Assay is a "qualitative in vitro diagnostic test" and an "automated real-time polymerase chain reaction (PCR) in vitro diagnostic test".

The intended use also clearly describes it as a test performed in vitro (outside the body) on biological specimens (urine, swabs) to aid in the diagnosis of a medical condition (trichomoniasis).

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The Cepheid Xpert TV Assay, performed on the GeneXpert® Instrument Systems, is a qualitative in vitro diagnostic test for the detection of Trichomonas vaginalis genomic DNA. The test utilizes automated real-time polymerase chain reaction (PCR) to detect Trichomonas vaginalis genomic DNA. The Xpert TV Assay uses female and male urine specimens, endocervical swab specimens, and patient-collected vaginal swab specimens (collected in a clinical setting). The Xpert TV Assay is intended to aid in the diagnosis of trichomoniasis in symptomatic or asymptomatic individuals.

Ancillary Collection Kits:

Xpert Vaginal/Endocervical Specimen Collection Kit
The Cepheid® Xpert® Vaginal/Endocervical Specimen Collection Kit is designed to collect, preserve, and transport Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis DNA in endocervical swab specimens (collected by a clinician) and patient-collected vaginal swab specimens (collected in a clinical setting) from symptomatic and asymptomatic women prior to analysis with the Xpert CT/NG Assay and the Xpert TV Assay.

Xpert Urine Specimen Collection Kit
The Cepheid® Xpert® Urine Specimen Collection Kit is designed to preserve and transport Chlamydia trachomatis. Neisseria gonorrhoeae, and Trichomonas vaginalis DNA in first-catch female urine specimens from symptomatic and asymptomatic individuals prior to analysis with the Xpert CT/NG Assay and the Xpert TV Assay.

Product codes

OUY, OOI, JSM

Device Description

The Xpert TV Assay is an automated real-time polymerase chain reaction (PCR) in vitro diagnostic test for qualitative detection of genomic DNA from Trichomonas vaginalis. The Xpert TV Assay is intended as an aid in the diagnosis of trichomoniasis.

The Xpert TV Assay is performed on the Cepheid GeneXpert® Instrument Systems (GeneXpert Dx., GeneXpert Infinity-48, GeneXpert Infinity-48s, and GeneXpert Infinity-80 systems). The GeneXpert Instrument System platform automates sample preparation, amplification and real-time detection.

The GeneXpert Instrument Systems require the use of single-use, disposable cartridges (the Xpert TV 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 TV Assay cartridges contain reagents for the detection of genomic DNA from T. vaginalis for use with the following specimens collected from symptomatic and asymptomatic individuals: female and male urine, endocervical swab and patient-collected vaginal swab (collected in a clinical setting). A Sample Processing Control (SPC), Sample Adequacy Control (SAC), and a Probe Check Control (PCC) are controls utilized by the GeneXpert Instrument System platform. The SPC, SAC, and PCC are controls utilized by the GeneXpert Instrument System platform. The SPC is present to control for adequate processing of the target trichomonads and to monitor the presence of inhibitors in the real-time PCR reaction to reduce the possibility of false negative results. The SAC reagents detect the presence of a single copy human gene and monitor whether the specimen contains human cells. The PCC verifies reagent rehydration, real-time 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 PCR for the detection of genomic DNA from T. vaginalis in 70 minutes or less. The GeneXpert Instrument Systems, comprised of the GeneXpert Dx Systems and the GeneXpert Infinity Systems, have 1 to 80 randomly accessible modules, depending upon the instrument, that are each 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 real-time PCR and RT-PCR and detection.

The swab and/or urine specimens are collected from asymptomatic or symptomatic patients and placed into a specimen transport tube containing preservative. The specimen is transferred to the sample chamber of the disposable fluidic cartridge (the Xpert TV cartridge). The user initiates a test from the system user interface and places the cartridge into the GeneXpert instrument platform, which performs hands-off real-time, multiplex PCR for detection of DNA. The results are automatically generated at the end of the process in a report that can be viewed and printed.

The ancillary specimen collection kits for use with the Xpert TV Assay are the Cepheid Xpert Vaginal/Endocervical Specimen Collection Kit and the Cepheid Xpert Urine Specimen Collection Kit.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Urogenital tract (urine, endocervical swab, vaginal swab, prostatic fluid)

Indicated Patient Age Range

The average age among eligible female study participants was 33.5 years (range = 18 to 78 years). The average age among eligible male study participants was 36.2 years (range = 16 to 78 years).

Intended User / Care Setting

Operators in Moderate and High Complexity labs

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 participants included consenting asymptomatic and symptomatic, sexually active males and females seen at locations including, but not limited to: OB/GYN, sexually transmitted disease (STD), and family planning clinics. The study specimens consisted of prospectively collected male urine, endocervical swabs, and patient-collected vaginal swabs (collected in a clinical setting). Clinician-collected vaginal swabs were collected for testing by the reference NAAT test and culture. Samples were collected from 17 clinical sites and tested at 11 sites. Reference testing was performed at 3 central laboratories.

A study participant was considered to be infected by PIS if either of the two reference test results were positive. The subject was considered to be not infected by PIS when both reference test results were negative.

Performance of the Xpert TV Assay was calculated relative to the PIS for each of the three female specimen types (endocervical swabs, patient-collected vaginal swabs and urine) or to the PIS for male urine, respectively.

Specimens with discrepant results between the Xpert TV Assay and the PIS were analyzed by validated bi-directional Sanger sequencing and results are footnoted in Table 5-10 for informational purposes only.

Total tests performed are 10,017, with 190 initial ERROR, INVALID, or NO RESULT outcomes. 167 specimens yielded valid results upon repeat assay. The overall valid reporting rate of the assay was 99.8% (9994/10,017).

Reproducibility Study:
Intra-site reproducibility was evaluated at three sites (two external, one in-house). Site 1 used an Infinity-80 instrument. Sites 2 and 3 used GeneXpert Dx instruments. Specimens were created by spiking Trichomonas vaginalis (ATCC® 30001™) into pooled, Trichomonas vaginalis negative urine (patient urine added to Cepheid Urine Transport Reagent) or vaginal swab matrix (vaginal swabs collected into Cepheid Swab Transport Reagent). The specimens were prepared at concentration levels representing high negative (below LoD), LoD (~1X LoD), moderate positive (~3X LoD), and negative (Trichomonas vaginalis negative clinical matrix). A panel of 8 specimens (4 in urine and 4 in vaginal swab matrix) was tested twice per day, on 12 different days, by two different operators, at each of three sites (8 specimens x 2 replicates x 12 days x 2 operators x 3 sites = 1,152 observations total). Three lots of Xpert TV Assay cartridges were used at each of the 3 testing sites, with each lot used for 4 days of testing. Positive and negative controls were included in the study. The Xpert TV Assay was performed according to the Xpert TV Assay procedure.

Instrument System Precision:
An in-house precision study was conducted to compare the performance of the GeneXpert Dx and the GeneXpert Infinity Instrument Systems using specimens comprised of Trichomonas vaginalis (ATCC® 30001™) spiked into negative urine (patient urine added to Cepheid Urine Transport Reagent) or vaginal swab matrix (vaginal swabs collected into Cepheid Swab Transport Reagent). The specimens were prepared at concentration levels representing high negative (below LoD), LoD (~1X LoD), moderate positive (~3X LoD), and negative (Trichomonas vaginalis negative clinical matrix). A panel of 8 specimens (4 in urine matrix and 4 in vaginal swab matrix) was tested on 12 different days by two operators. Each operator conducted four runs of each panel specimen per day on each of the three instrument systems (8 specimens x 4 times/day x 12 days x 2 operators x 3 instrument systems = 2,304 observations total). Three lots of Xpert TV Assay cartridges were used for the study, with each lot used for 4 days of testing. Positive and negative controls were included in the study. The Xpert TV Assay was performed according to the Xpert TV Assay procedure.

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

Clinical Performance: Multi-site prospective investigational study.
The study included 10,017 tests.
Female: 1867 subjects
Male: 4626 subjects
Results of the Xpert TV Assay were compared to the Patient Infected Status (PIS) for determination of sensitivity, specificity, and predictive values.
Key Results are provided under Key Metrics.

Reproducibility Study: Intra-site reproducibility study, 1,152 observations total.
Female Swab Matrix (FS):
FS-Neg: 100% agreement (144/144)
FS-Mod Pos (~3X LoD; ~6 cells/mL): 100% agreement (144/144)
FS-LoD (~1X LoD; ~2 cells/mL): 95.8% agreement (138/144)
FS-High Neg (below LoD;

Regulation Number and Section

§ 866.3860

Trichomonas vaginalis nucleic acid assay.(a)
Identification. ATrichomonas vaginalis nucleic acid assay is a device that consists of primers, probes, enzymes, and controls for the amplification and detection of trichomonas nucleic acids in endocervical swabs, vaginal swabs, and female urine specimens, from women symptomatic for vaginitis, cervicitis, or urethritis and/or to aid in the diagnosis of trichomoniasis in asymptomatic women. The detection of trichomonas nucleic acids, in conjunction with other laboratory tests, aids in the clinical laboratory diagnosis of trichomoniasis caused byTrichomonas vaginalis .(b)
Classification. Class II (special controls). The special controls are set forth in FDA's guideline document entitled: “Class II Special Controls Guideline: Nucleic Acid Amplification Assays for the Detection ofTrichomonas vaginalis; Guideline for Industry and Food and Drug Administration Staff.” See § 866.1(e) for information on obtaining this document.

Summary

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Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health and Human Services. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter. Inside the circle is a stylized image of three human profiles facing right, resembling a bird in flight.

August 29, 2016

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

Cepheid Scott Campbell, Ph.D., MBA Corporate Vice President and Chief Regulatory Officer 904 Caribbean Drive Sunnyvale, CA 94089-1189

Re: K161619

Trade/Device Name: Xpert® TV, Cepheid GeneXpert® Instrument Systems (GeneXpert Dx, GeneXpert Infinity-48, GeneXpert Infinity-48s, and GeneXpertInfinity-80 systems). Xpert® Vaginal/Endocervical Specimen Collection Kit and Xpert® Urine Specimen Collection Kit. Regulation Number: 21 CFR 866.3860 Regulation Name: Trichomonas vaginalis nucleic acid assay

Regulatory Class: II Product Code: OUY, OOI, JSM Dated: June 10, 2016 Received: June 15, 2016

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

1

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.

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,

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

2

Indications for Use

510(k) Number (if known) K161619

Device Name Xpert® TV

Indications for Use (Describe)

The Cepheid Xpert TV Assay, performed on the GeneXpert® Instrument Systems, is a qualitative in vitro diagnostic test for the detection of Trichomonas vaginalis genomic DNA. The test utilizes automated real-time polymerase chain reaction (PCR) to detect Trichomonas vaginalis genomic DNA. The Xpert TV Assay uses female and male urine specimens, endocervical swab specimens, and patient-collected vaginal swab specimens (collected in a clinical setting). The Xpert TV Assay is intended to aid in the diagnosis of trichomoniasis in symptomatic or asymptomatic individuals.

Ancillary Collection Kits:

Xpert Vaginal/Endocervical Specimen Collection Kit

The Cepheid® Xpert® Vaginal/Endocervical Specimen Collection Kit is designed to collect, preserve, and transport Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis DNA in endocervical swab specimens (collected by a clinician) and patient-collected vaginal swab specimens (collected in a clinical setting) from symptomatic and asymptomatic women prior to analysis with the Xpert CT/NG Assay and the Xpert TV Assay.

Xpert Urine Specimen Collection Kit

The Cepheid® Xpert® Urine Specimen Collection Kit is designed to preserve and transport Chlamydia trachomatis. Neisseria gonorrhoeae, and Trichomonas vaginalis DNA in first-catch female urine specimens from symptomatic and asymptomatic individuals prior to analysis with the Xpert CT/NG Assay and the Xpert TV 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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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: | July 13, 2016 |
| Device: | |
| Trade name: | Xpert® TV |
| Common name: | Xpert TV Assay |
| Type of Test: | Real-Time Polymerase Chain Reaction (PCR) for the detection
of Trichomonas vaginalis |
| Regulation number/
Classification name/ | 866.3860/ Trichomonas vaginalis nucleic acid amplification test
system /OUY |
| Product code: | 862.2570/Instrumentation for clinical multiplex test
systems/OOI |
| Classification
Advisory Panel | Microbiology (83) |
| Prescription Use | Yes |
| Predicate Device
Assay: | Cepheid Xpert TV Assay
[510(k) #K151565] |
| Predicate Device
Assay: | Xpert Urine Specimen Collection Kit
[510(k) #K151565] |

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Device Description:

The Xpert TV Assay is an automated real-time polymerase chain reaction (PCR) in vitro diagnostic test for qualitative detection of genomic DNA from Trichomonas vaginalis. The Xpert TV Assay is intended as an aid in the diagnosis of trichomoniasis.

The Xpert TV Assay is performed on the Cepheid GeneXpert® Instrument Systems (GeneXpert Dx., GeneXpert Infinity-48, GeneXpert Infinity-48s, and GeneXpert Infinity-80 systems). The GeneXpert Instrument System platform automates sample preparation, amplification and real-time detection.

The GeneXpert Instrument Systems require the use of single-use, disposable cartridges (the Xpert TV 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 TV Assay cartridges contain reagents for the detection of genomic DNA from T. vaginalis for use with the following specimens collected from symptomatic and asymptomatic individuals: female and male urine, endocervical swab and patientcollected vaginal swab (collected in a clinical setting). A Sample Processing Control (SPC), Sample Adequacy Control (SAC), and a Probe Check Control (PCC) are controls utilized by the GeneXpert Instrument System platform. The SPC, SAC, and PCC are controls utilized by the GeneXpert Instrument System platform. The SPC is present to control for adequate processing of the target trichomonads and to monitor the presence of inhibitors in the real-time PCR reaction to reduce the possibility of false negative results. The SAC reagents detect the presence of a single copy human gene and monitor whether the specimen contains human cells. The PCC verifies reagent rehydration, real-time 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 PCR for the detection of genomic DNA from T. vaginalis in 70 minutes or less. The GeneXpert Instrument Systems, comprised of the GeneXpert Dx Systems and the GeneXpert Infinity Systems, have 1 to 80 randomly accessible modules, depending upon the instrument, that are each 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 real-time PCR and RT-PCR and detection.

The swab and/or urine specimens are collected from asymptomatic or symptomatic patients and placed into a specimen transport tube containing preservative. The specimen is transferred to the sample chamber of the disposable fluidic cartridge (the Xpert TV cartridge). The user initiates a test from the system user interface and places the cartridge

1 Although sonication is a fundamental capability of every GeneXpert module, sonication is not used in the Xpert TV Assay.

5

into the GeneXpert instrument platform, which performs hands-off real-time, multiplex PCR for detection of DNA. The results are automatically generated at the end of the process in a report that can be viewed and printed.

The ancillary specimen collection kits for use with the Xpert TV Assay are the Cepheid Xpert Vaginal/Endocervical Specimen Collection Kit and the Cepheid Xpert Urine Specimen Collection Kit.

Device Intended Use:

The Cepheid Xpert TV Assay, performed on the GeneXpert Instrument Systems, is a qualitative in vitro diagnostic test for the detection of Trichomonas vaginalis genomic DNA. The test utilizes automated real-time polymerase chain reaction (PCR) to detect Trichomonas vaginalis genomic DNA. The Xpert TV Assay uses female and male urine specimens, endocervical swab specimens, and patient-collected vaginal swab specimens (collected in a clinical setting). The Xpert TV Assay is intended to aid in the diagnosis of trichomoniasis in symptomatic or asymptomatic individuals.

Ancillary Collection Kits:

Xpert Vaginal/Endocervical Specimen Collection Kit

The Cepheid® Xpert® Vaginal/Endocervical Specimen Collection Kit is designed to collect, preserve, and transport Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis DNA in endocervical swab specimens (collected by a clinician) and patient-collected vaginal swab specimens (collected in a clinical setting) from symptomatic and asymptomatic women prior to analysis with the Xpert CT/NG Assay and the Xpert TV Assay.

Xpert Urine Specimen Collection Kit

The Cepheid® Xpert® Urine Specimen Collection Kit is designed to preserve and transport Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis DNA in first-catch female and male urine specimens from symptomatic and asymptomatic individuals prior to analysis with the Xpert CT/NG Assay and the Xpert TV Assay.

Substantial Equivalence:

The Xpert TV Assay is substantially equivalent to the cleared Cepheid Xpert TV Assay [510(k) #K151565]. The assays are identical in reagents and formulation and use the same real-time PCR technology and GeneXpert instruments. The performance of the Xpert TV Assay was evaluated in a multi-site clinical study in which the performance of the Xpert TV Assay was compared to a patient-infected status (PIS). The results of the study demonstrated that the performance of the Xpert TV Assay is substantially equivalent to the predicate device.

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Table 5-1 shows the similarities and differences between the Xpert TV Assay and the predicate device.

Similarities
Item	Device	Predicate Device
	Cepheid Xpert TV Assay	Current Cepheid
Xpert TV Assay
510(k) Number	K161619	K151565
Regulation	Same	866.3860
Product Code	Same	OUY
Device Class	Same	II
Intended Use	The Cepheid Xpert TV Assay,
performed on the GeneXpert®
Instrument Systems, is a
qualitative in vitro diagnostic
test for the detection of
Trichomonas vaginalis
genomic DNA. The test
utilizes automated real-time
polymerase chain reaction
(PCR) to detect Trichomonas
vaginalis genomic DNA. The
Xpert TV Assay uses female
and male urine specimens,
endocervical swab specimens,
or patient-collected vaginal
swab specimens (collected in a
clinical setting). The Xpert
TV Assay is intended to aid in
the diagnosis of trichomoniasis
in symptomatic or
asymptomatic individuals.	The Cepheid Xpert TV Assay,
performed on the GeneXpert®
Instrument Systems, is a
qualitative in vitro diagnostic
test for the detection of
Trichomonas vaginalis
genomic DNA. The test
utilizes automated real-time
polymerase chain reaction
(PCR) to detect Trichomonas
vaginalis genomic DNA. The
Xpert TV Assay uses female
urine specimens, endocervical
swab specimens, or patient-
collected vaginal swab
specimens (collected in a
clinical setting). The Xpert
TV Assay is intended to aid in
the diagnosis of trichomoniasis
in symptomatic or
asymptomatic individuals.
Assay Targets	Same	T. vaginalis genomic DNA
Specimen Types	Endocervical Swabs
Vaginal Swabs
Female Urine
Male Urine	Endocervical Swabs
Vaginal Swabs
Female Urine

Table 5-1: Comparison of Similarities and Differences of the
Xpert TV Assay with the Predicate Device

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Similarities
Item	Device	Predicate Device
	Cepheid Xpert TV Assay	Current Cepheid
Xpert TV Assay
Nucleic Acid
Extraction	Yes	Yes
Assay Results	Same	Qualitative
Collection Kit	Same	Urine collection kit
Swab collection kit
Technology/
Detection	Same	Real-time polymerase chain
reaction (PCR)
Instrument System	Same	Cepheid GeneXpert
Instrument System
Laboratory Users	Same	Operators in Moderate and
High Complexity labs
Early assay
termination function	Same	Yes
(for positive samples)

Primary Differences
	New Device	Predicate Device
Item	Cepheid Xpert TV Assay	Current Cepheid
Xpert TV Assay
Specimen Types	Endocervical Swabs
Vaginal Swabs
Female Urine
Male Urine	Endocervical Swabs
Vaginal Swabs
Female Urine

The Xpert TV 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 TV Assay and the predicate device do not raise different questions of safety and effectiveness. The clinical study demonstrates that the Xpert TV Assay is acceptable for its intended use with inexperienced lab users and is substantially equivalent to the predicate device described above.

8

Ancillary Collection Kit:

Xpert Urine Specimen Collection Kit

The predicate device for the Cepheid Xpert Urine Specimen Collection Kit is the current Cepheid Xpert Urine Specimen Collection Kit [510(k) # K151565]. The similarities and differences are shown in Table 5-2.

Similarities
Item	Device
Cepheid Xpert Urine
Specimen Collection Kit	Predicate
Current Cepheid Xpert Urine
Specimen Collection Kit
Intended Use
(Similarities)	The collection kit is designed
to preserve and transport
Chlamydia trachomatis ,
Neisseria gonorrhoeae and
Trichomonas vaginalis DNA in
first-catch male and female
urine specimens from
symptomatic and
asymptomatic individuals prior
to analysis.	The collection kit is designed
to preserve and transport
Chlamydia trachomatis and
Neisseria gonorrhoeae and
Trichomonas vaginalis DNA in
first-catch female urine
specimens from symptomatic
and asymptomatic individuals
prior to analysis.
Single-use Device	Yes	Yes
Transport Medium
pH	Same	7.95 – 8.35
Description	Same	Contains one individually
packaged sterile disposable
transfer pipette and one urine
transport reagent tube.
Approximately 7 mL of a first-
catch urine specimen is
transferred to the Urine
Transport Reagent tube to
preserve and transport the
specimen prior to analysis with
the assay.

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

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Differences
	Device	Predicate
Item	Cepheid Xpert Urine
Specimen Collection Kit	Current Cepheid Xpert Urine
Specimen Collection Kit
Intended Use
(differences)	Specimen storage and transport
when testing for the presence of
Chlamydia trachomatis and
Neisseria gonorrhoeae in first-
catch male and female urine
specimens and for the presence
of Trichomonas vaginalis in
first-catch female and male
urine specimens.	Specimen storage and transport
when testing for the presence of
Chlamydia trachomatis and
Neisseria gonorrhoeae in first-
catch male and female urine
specimens and for the presence
of Trichomonas vaginalis in
first-catch female urine
specimens.

Non-Clinical Studies:

Analytical Sensitivity (Limit of Detection)

The analytical sensitivity or limit of detection (LoD) of the Xpert TV Assay was assessed using two Trichomonas vaginalis strains, one metronidazole susceptible (T. vaginalis ATCC® 30001™), and one metronidazole resistant (T. vaginalis ATCC® 30238"). The strains were tested individually in clinical T. vaginalis-negative pooled urine matrix in Cepheid Xpert Urine Transport Reagent and clinical T. vaginalisnegative pooled vaginal swab matrix (VS) in Cepheid Xpert Swab Transport Reagent.

T. vaginalis was cultured and incubated at 35°C. Visual examination of the cultures for white precipitate (indicating growth) was conducted every 24 hours for 3 to 5 days. Cell pellets were resuspended in growth medium and enumerated visually using light microscopy. The concentration of isolates was expressed as the number of cells per milliliter (cells/mL). Cultures were diluted in culture medium to 1 x10+ cells/mL and stored at -20°C. Cells were thawed on ice for use in the study.

The LoD was estimated by testing replicates of 20 at five concentrations for each strain and sample type over three days. The LoD for each strain was estimated by probit analysis. The claimed LoDs were confirmed by analyzing at least 20 replicates with T. vaginalis cells diluted to the estimated LoD concentrations. The LoD is defined as the lowest number of cells/mL that can be reproducibly distinguished from negative samples with 95% confidence or the lowest concentration at which 19 of 20 replicates were positive. The study was performed with two different lots of Xpert TV reagents and the claimed LoD for each strain is the higher of the two determinations (Table 5-3). The claimed LoD for T. vaginalis strains ATCC 30001 and ATCC 30238 in vaginal swab matrix is 2 cells/mL. The claimed LoD for T. vaginalis strain ATCC 30001 in urine matrix is 3 cells/mL. The claimed LoD for T. vaginalis strain ATCC 30238 in

10

urine matrix is 2 cells/mL.

The LoD point estimates and confirmed LoD for each strain tested are summarized in Table 5-3.

| Trichomonas
vaginalis | LoD Estimates by
Probit Analysis
(cells/mL) | | Verified
LoD
(cells/mL) | Verification
(Positives/20) | Mean TV
Ct | Mean SAC
Ct | Mean SPC
Ct | LoD
Claim
(cells/mL) |
|-------------------------------|---------------------------------------------------|------------------|-------------------------------|--------------------------------|---------------|----------------|----------------|----------------------------|
| strain and matrix | Reagent
Lot 1 | Reagent
Lot 2 | | | | | | |
| ATCC 30001 in
Vaginal Swab | 2.0 | 1.6 | 2.0 | 20/20 | 39.1 | 21.4 | 33.9 | 2 |
| ATCC 30238 in
Vaginal Swab | 1.7 | 2.1 | 2.1 | 20/20 | 37.5 | 21.4 | 33.7 | 2 |
| ATCC 30001 in
Urine | 2.2 | 2.5 | 2.5 | 20/20 | 38.2 | 29.3 | 34.1 | 3 |
| ATCC 30238 in
Urine | 2.1 | 1.7 | 2.1 | 20/20 | 38.2 | 29.2 | 33.8 | 2 |

Table 5-3: LoD of Two T. vaginalis Strains in Pooled Vaginal Swab Matrix and Urine Matrix

Analytical Specificity (Cross-reactivity and Competitive Interference)

A panel of 124 microorganisms, including bacteria, fungi, and viruses commonly found in the urogenital tract, as well as other protozoans closely related to T. vaginalis were tested with the Xpert TV Assay. The microorganisms were tested in the presence (competitive interference) and absence (cross-reactivity) of 3X LoD T. vaginalis ATCC 30001 cells. The microorganisms were seeded into either pooled Trichomonas vaginalis-negative urine matrix (patient urine added to Cepheid Urine Transport Reagent) or pooled Trichomonas vaginalis-negative vaginal swab matrix (vaginal swabs collected into Cepheid Swab Transport Reagent).

Each bacterial or fungal strain was tested at 1 x 10°CFU/mL or greater or at 1 x 10° genomes/mL. Viral strains were tested at 1 x 105 TCIDs0/mL or 105 genomes/mL or greater. Protozoans were cultured in growth media, visually enumerated by light microscopy and tested at 1 x 105 cells/mL or greater or 105 genomes/mL. All microorganisms were tested in triplicate. Positive and negative controls were included in the study. One organism, Trichomonas tenax, demonstrated cross-reactivity (result of TV DETECTED in the absence of TV) at 1 x 10 cells/mL for the urine and vaginal swab matrix samples. Trichomonas tenax was subjected to repeat analysis at various other concentrations until a result of TV NOT DETECTED was obtained (at 1 x 10- cells/mL). This is addressed in Limitations in the package insert. For the other 123 microorganisms. all TV positive samples remained positive and all TV negative samples remained negative, indicating that there was no interference or cross-reactivity with the results of the Xpert TV Assay for these microorganisms. Results are shown in Table 5-4 and Table 5-5 for urine and vaginal swab matrix, respectively.

11

| Microorganism Testeda | Cross-Reactivity
(- T. vaginalis) | Competitive
Interference
(+ T. vaginalis) | |---------------------- | Achromobacter xerosis | Acinetobacter calcoaceticus | Acinetobacter lwoffii | Actinomyces israeliib | Actinomyces pyogenes | Aerococcus viridans | Aeromonas hydrophila | Alcaligenes faecalis | Atopobium vaginaeb | Bacillus subtilis | Bacteroides fragilisb | Bacteroides ureolyticusb | Bifidobacterium adolescentisb | Bifidobacterium | Blastocystis hominisc | Branhamella catarrhalis | Brevibacterium linens | Campylobacter jejuni | Candida albicanse | Candida glabratae | Candida parapsilosie | Candida tropicalise | Chlamydia trachomatis | Chromobacterium violaceum | Citrobacter freundii | Clostridium difficileb | Clostridium perfringensb | Corynebacterium genitalium | Corynebacterium xerosis | Microorganism Testeda | (- T. vaginalis) | Competitive
Interference
(+ T. vaginalis) | | Cryptococcus neoformanse | Cryptosporidium parvumc | Cytomegalovirusf | Deinococcus radiodurans | Derxia gummosa | Eikenella corrodens | Entamoeba histolyticac | Enterobacter aerogenes | Enterobacter cloacae | Enterococcus avium | Enterococcus faecalis | Enterococcus faecium | Erysipelothrix rhusiopathiae | Escherichia coli | Flavobacterium | Fusobacterium nucleatumb | Gardnerella vaginalis | Gemella haemolysans | Giardia intestinalisc | Haemophilus ducreyi | Haemophilus influenzae | Herpes simplex virus If | Herpes simplex virus IIf | HIV-1f | Human papilloma virus 16f | Kingella dentrificans | Kingella kingae | Klebsiella oxytoca | Klebsiella pneumoniae | Lactobacillus acidophilus | Lactobacillus brevis | Lactobacillus crispatus | Microorganism Testeda | (- T. vaginalis) | Competitive
Interference
(+ T. vaginalis) | | Lactobacillus jensonii | Lactobacillus lactis | Lactobacillus vaginalis | Legionella pneumophila | Leuconostoc | Listeria monocytogenes | Micrococcus luteus | Mobiluncus curtisiib | Moraxella lacunata | Moraxella osloensis | Morganella morganii | Mycobacterium smegmatis | Mycoplasma genitalium | Mycoplasma hominis | Neisseria cinerea | Neisseria dentrificans | Neisseria elongata | Neisseria flava | Neisseria flavescens | Neisseria gonorrhoeae | Neisseria lactamica | Neisseria mucosa | Neisseria perflava | Neisseria polysaccharea | Neisseria sicca | Neisseria subflava | Pantoea agglomerans | Paracoccus denitrificans | Pentatrichomonis hominisc | Peptostreptococcus | Peptostreptococcus productusb | Plesiomonas shigelloides | Microorganism Testeda | (- T. vaginalis) | Competitive
Interference
(+ T. vaginalis) | | Prevotella biviab | Propionibacterium acnesb | Proteus mirabilis | Proteus vulgaris | Providencia stuartii | Pseudomonas aeruginosa | Pseudomonas fluorescens | Pseudomonas putida | Rahnella aquatilis | Rhodospirillum rubrum | Saccharomyces cerevisiaee | Salmonella minnesota | Salmonella typhimurium | Serratia marcescens | Staphylococcus aureus | Staphylococcus epidermidis | Staphylococcus saprophyticus | Streptococcus agalactiae | Streptococcus bovis | Streptococcus mitis | Streptococcus mutans | Streptococcus pneumoniae | Streptococcus pyogenes | Streptococcus salivarius | Streptococcus sanguis | Streptomyces griseinus | Trichomonas tenaxc | Trichomonas tenaxc | Trichomonas tenaxc | Ureaplasma parvum | Ureaplasma urealyticum | Vibrio parahaemolyticus | Testeda | (- T. vaginalis ) | Competitive
Interference
(+ T. vaginalis ) |
| | Concentration
|
----------|--------------------------------|--------------------------------------|-------------------------------------------------|---------------------------------------------------------|
| 8 x 106 | TV NOT DETECTED | TV DETECTED | |
| 3 x 106 | TV NOT DETECTED | TV DETECTED | |
| 6 x 106 | TV NOT DETECTED | TV DETECTED | |
| 2 x 106 | TV NOT DETECTED | TV DETECTED | |
| 8 x 106 | TV NOT DETECTED | TV DETECTED | |
| 5 x 106 | TV NOT DETECTED | TV DETECTED | |
| 1 x 107 | TV NOT DETECTED | TV DETECTED | |
| 1 x 107 | TV NOT DETECTED | TV DETECTED | |
| 2 x 106 | TV NOT DETECTED | TV DETECTED | |
| 6 x 106 | TV NOT DETECTED | TV DETECTED | |
| 5 x 106 | TV NOT DETECTED | TV DETECTED | |
| 1 x 106 | TV NOT DETECTED | TV DETECTED | |
| 6 x 106 | TV NOT DETECTED | TV DETECTED | |
brevi (breve)b | 9 x 106 | TV NOT DETECTED | TV DETECTED | |
| 1 x 105d | TV NOT DETECTED | TV DETECTED | |
| 1 x 107 | TV NOT DETECTED | TV DETECTED | |
| 7 x 106 | TV NOT DETECTED | TV DETECTED | |
| 1 x 107 | TV NOT DETECTED | TV DETECTED | |
| 7 x 106 | TV NOT DETECTED | TV DETECTED | |
| 4 x 106 | TV NOT DETECTED | TV DETECTED | |
| 7 x 106 | TV NOT DETECTED | TV DETECTED | |
| 6 x 106 | TV NOT DETECTED | TV DETECTED | |
| 2 x 106 | TV NOT DETECTED | TV DETECTED | |
| 7 x 106 | TV NOT DETECTED | TV DETECTED | |
| 1 x 106 | TV NOT DETECTED | TV DETECTED | |
| 4 x 106 | TV NOT DETECTED | TV DETECTED | |
| 2 x 106 | TV NOT DETECTED | TV DETECTED | |
| 2 x 106 | TV NOT DETECTED | TV DETECTED | |
| 3 x 106 | TV NOT DETECTED | TV DETECTED | |
| Concentration
| Xpert TV Assay Result | | |
| | Cross-Reactivity
|
| 6 x 106 | TV NOT DETECTED | TV DETECTED | |
| 1 x 105d | TV NOT DETECTED | TV DETECTED | |
| 5 x 105 | TV NOT DETECTED | TV DETECTED | |
| 5 x 106 | TV NOT DETECTED | TV DETECTED | |
| 1 x 105d | TV NOT DETECTED | TV DETECTED | |
| 5 x 106 | TV NOT DETECTED | TV DETECTED | |
| 1 x 105d | TV NOT DETECTED | TV DETECTED | |
| 1 x 106 | TV NOT DETECTED | TV DETECTED | |
| 2 x 106 | TV NOT DETECTED | TV DETECTED | |
| 7 x 106 | TV NOT DETECTED | TV DETECTED | |
| 7 x 106 | TV NOT DETECTED | TV DETECTED | |
| 4 x 106 | TV NOT DETECTED | TV DETECTED | |
| 1 x 107 | TV NOT DETECTED | TV DETECTED | |
| 1 x 107 | TV NOT DETECTED | TV DETECTED | |
meningosepticum | 1 x 107 | TV NOT DETECTED | TV DETECTED | |
| 4 x 106 | TV NOT DETECTED | TV DETECTED | |
| 2 x 106 | TV NOT DETECTED | TV DETECTED | |
| 6 x 106 | TV NOT DETECTED | TV DETECTED | |
| 1 x 105d | TV NOT DETECTED | TV DETECTED | |
| 1 x 105d | TV NOT DETECTED | TV DETECTED | |
| 3 x 106 | TV NOT DETECTED | TV DETECTED | |
| 1 x 105 | TV NOT DETECTED | TV DETECTED | |
| 1 x 105 | TV NOT DETECTED | TV DETECTED | |
| 2 x 105 | TV NOT DETECTED | TV DETECTED | |
| 6 x 105 | TV NOT DETECTED | TV DETECTED | |
| 3 x 106 | TV NOT DETECTED | TV DETECTED | |
| 1 x 107 | TV NOT DETECTED | TV DETECTED | |
| 1 x 107 | TV NOT DETECTED | TV DETECTED | |
| 7 x 106 | TV NOT DETECTED | TV DETECTED | |
| 8 x 106 | TV NOT DETECTED | TV DETECTED | |
| 7 x 106 | TV NOT DETECTED | TV DETECTED | |
| 2 x 106 | TV NOT DETECTED | TV DETECTED | |
| Concentration
| Xpert TV Assay Result | | |
| | Cross-Reactivity
|
| 8 x 106 | TV NOT DETECTED | TV DETECTED | |
| 7 x 106 | TV NOT DETECTED | TV DETECTED | |
| 6 x 106 | TV NOT DETECTED | TV DETECTED | |
| 5 x 106 | TV NOT DETECTED | TV DETECTED | |
paramensenteroides | 7 x 106 | TV NOT DETECTED | TV DETECTED | |
| 1 x 107 | TV NOT DETECTED | TV DETECTED | |
| 3 x 106 | TV NOT DETECTED | TV DETECTED | |
| 3 x 106 | TV NOT DETECTED | TV DETECTED | |
| 6 x 106 | TV NOT DETECTED | TV DETECTED | |
| 5 x 106 | TV NOT DETECTED | TV DETECTED | |
| 1 x 107 | TV NOT DETECTED | TV DETECTED | |
| 7 x 106 | TV NOT DETECTED | TV DETECTED | |
| 1 x 106d | TV NOT DETECTED | TV DETECTED | |
| 1 x 106d | TV NOT DETECTED | TV DETECTED | |
| 1 x 106 | TV NOT DETECTED | TV DETECTED | |
| 2 x 106 | TV NOT DETECTED | TV DETECTED | |
| 7 x 106 | TV NOT DETECTED | TV DETECTED | |
| 9 x 106 | TV NOT DETECTED | TV DETECTED | |
| 8 x 106 | TV NOT DETECTED | TV DETECTED | |
| 6 x 106 | TV NOT DETECTED | TV DETECTED | |
| 3 x 106 | TV NOT DETECTED | TV DETECTED | |
| 9 x 106 | TV NOT DETECTED | TV DETECTED | |
| 3 x 106 | TV NOT DETECTED | TV DETECTED | |
| 4 x 106 | TV NOT DETECTED | TV DETECTED | |
| 8 x 106 | TV NOT DETECTED | TV DETECTED | |
| 9 x 106 | TV NOT DETECTED | TV DETECTED | |
| 4 x 106 | TV NOT DETECTED | TV DETECTED | |
| 8 x 106 | TV NOT DETECTED | TV DETECTED | |
| 1 x 106 | TV NOT DETECTED | TV DETECTED | |
anaerobiusb | 4 x 106 | TV NOT DETECTED | TV DETECTED | |
| 2 x 106 | TV NOT DETECTED | TV DETECTED | |
| 4 x 106 | TV NOT DETECTED | TV DETECTED | |
| Concentration
| Xpert TV Assay Result | | |
| | Cross-Reactivity
|
| 1 x 106 | TV NOT DETECTED | TV DETECTED | |
| 3 x 106 | TV NOT DETECTED | TV DETECTED | |
| 1 x 107 | TV NOT DETECTED | TV DETECTED | |
| 1 x 107 | TV NOT DETECTED | TV DETECTED | |
| 1 x 107 | TV NOT DETECTED | TV DETECTED | |
| 8 x 106 | TV NOT DETECTED | TV DETECTED | |
| 8 x 106 | TV NOT DETECTED | TV DETECTED | |
| 7 x 106 | TV NOT DETECTED | TV DETECTED | |
| 1 x 107 | TV NOT DETECTED | TV DETECTED | |
| 1 x 106d | TV NOT DETECTED | TV DETECTED | |
| 6 x 106 | TV NOT DETECTED | TV DETECTED | |
| 1 x 106 | TV NOT DETECTED | TV DETECTED | |
| 1 x 107 | TV NOT DETECTED | TV DETECTED | |
| 2 x 106 | TV NOT DETECTED | TV DETECTED | |
| 9 x 106 | TV NOT DETECTED | TV DETECTED | |
| 7 x 106 | TV NOT DETECTED | TV DETECTED | |
| 8 x 106 | TV NOT DETECTED | TV DETECTED | |
| 1 x 107 | TV NOT DETECTED | TV DETECTED | |
| 8 x 106 | TV NOT DETECTED | TV DETECTED | |
| 2 x 106 | TV NOT DETECTED | TV DETECTED | |
| 2 x 107 | TV NOT DETECTED | TV DETECTED | |
| 1 x 106 | TV NOT DETECTED | TV DETECTED | |
| 2 x 106 | TV NOT DETECTED | TV DETECTED | |
| 9 x 106 | TV NOT DETECTED | TV DETECTED | |
| 5 x 106 | TV NOT DETECTED | TV DETECTED | |
| 6 x 106 | TV NOT DETECTED | TV DETECTED | |
| 1 x 105 | TV DETECTED | TV DETECTED | |
| 1 x 103 | TV DETECTED | TV DETECTED | |
| 1 x 102 | TV NOT DETECTED | TV DETECTED | |
| 1 x 106d | TV NOT DETECTED | TV DETECTED | |
| 1 x 106 | TV NOT DETECTED | TV DETECTED | |
| 9 x 106 | TV NOT DETECTED | TV DETECTED | |
| Microorganism | Concentration
| Xpert TV Assay Result | |
| | | Cross-Reactivity
| Yersinia enterocolitica | 8 x 106 | TV NOT DETECTED | TV DETECTED |

Table 5-4: Analytical Specificity/Competitive Interference Determination for Xpert TV Assay in Urine Matrix

12

13

14

15

a. Tests run ≥ 10° CFU/mL for bacteria and fungi, ≥10° genomes/mL for yeast, ≥10° TCIDsy/mL or ≥10° genomes/mL for viruses and ≥105 cells/mL for protozoans.

b. Anaerobic organism

c. Protozoan

d. Genome equivalents tested (DNA)

e. Fungal organism

f. Virus

Table 5-5: Analytical Specificity/Competitive Interference Determination for
Xpert TV Assav in Vaginal Swab Matrix

| Microorganism | Concentration
Testeda | Xpert TV Assay Result | |
|-----------------------------------|--------------------------|--------------------------------------|-------------------------------------------------|
| | | Cross-Reactivity
(- T. vaginalis) | Competitive
Interference
(+ T. vaginalis) |
| Achromobacter xerosis | 1 x 106 | TV NOT DETECTED | TV DETECTED |
| Acinetobacter calcoaceticus | 3 x 106 | TV NOT DETECTED | TV DETECTED |
| Acinetobacter lwoffii | 6 x 106 | TV NOT DETECTED | TV DETECTED |
| Actinomyces israeliib | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Actinomyces pyogenes | 8 x 106 | TV NOT DETECTED | TV DETECTED |
| Aerococcus viridans | 5 x 106 | TV NOT DETECTED | TV DETECTED |
| Aeromonas hydrophila | 5 x 106 | TV NOT DETECTED | TV DETECTED |
| Alcaligenes faecalis | 5 x 106 | TV NOT DETECTED | TV DETECTED |
| Atopobium vaginaeb | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Bacillus subtilis | 6 x 106 | TV NOT DETECTED | TV DETECTED |
| Bacteroides fragilisb | 5 x 106 | TV NOT DETECTED | TV DETECTED |
| Bacteroides ureolyticusb | 1 x 106 | TV NOT DETECTED | TV DETECTED |
| Bifidobacterium adolescentisb | 6 x 106 | TV NOT DETECTED | TV DETECTED |
| Bifidobacterium brevi (breve)b | 9 x 106 | TV NOT DETECTED | TV DETECTED |
| Blastocystis hominisc | 1 x 105 d | TV NOT DETECTED | TV DETECTED |
| Branhamella catarrhalis | 3 x 106 | TV NOT DETECTED | TV DETECTED |
| Brevibacterium linens | 8 x 106 | TV NOT DETECTED | TV DETECTED |
| Campylobacter jejuni | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Candida albicanse | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Candida glabratae | 4 x 106 | TV NOT DETECTED | TV DETECTED |
| Candida parapsilosie | 3 x 106 | TV NOT DETECTED | TV DETECTED |
| | Concentration
Testeda | Xpert TV Assay Result | |
| Microorganism | | Cross-Reactivity
(- T. vaginalis) | Competitive
Interference
(+ T. vaginalis) |
| Candida tropicalise | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Chlamydia trachomatis | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Chromobacterium violaceum | 7 x 106 | TV NOT DETECTED | TV DETECTED |
| Citrobacter freundii | 1 x 106 | TV NOT DETECTED | TV DETECTED |
| Clostridium difficileb | 4 x 106 | TV NOT DETECTED | TV DETECTED |
| Clostridium perfringensb | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Corynebacterium genitalium | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Corynebacterium xerosis | 3 x 106 | TV NOT DETECTED | TV DETECTED |
| Cryptococcus neoformanse | 5 x 106 | TV NOT DETECTED | TV DETECTED |
| Cryptosporidium parvumc | 1 x 105 d | TV NOT DETECTED | TV DETECTED |
| Cytomegalovirusf | 5 x 105 | TV NOT DETECTED | TV DETECTED |
| Deinococcus radiodurans | 1 x 106 | TV NOT DETECTED | TV DETECTED |
| Derxia gummosa | 1 x 106 d | TV NOT DETECTED | TV DETECTED |
| Eikenella corrodens | 8 x 106 | TV NOT DETECTED | TV DETECTED |
| Entamoeba histolyticac | 1 x 105 d | TV NOT DETECTED | TV DETECTED |
| Enterobacter aerogenes | 1 x 106 | TV NOT DETECTED | TV DETECTED |
| Enterobacter cloacae | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Enterococcus avium | 7 x 106 | TV NOT DETECTED | TV DETECTED |
| Enterococcus faecalis | 7 x 106 | TV NOT DETECTED | TV DETECTED |
| Enterococcus faecium | 3 x 106 | TV NOT DETECTED | TV DETECTED |
| Erysipelothrix rhusiopathiae | 3 x 106 | TV NOT DETECTED | TV DETECTED |
| Escherichia coli | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Flavobacterium
meningosepticum | 5 x 106 | TV NOT DETECTED | TV DETECTED |
| Fusobacterium nucleatumb | 4 x 106 | TV NOT DETECTED | TV DETECTED |
| Gardnerella vaginalis | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Gemella haemolysans | 9 x 106 | TV NOT DETECTED | TV DETECTED |
| Giardia intestinalisc | 1 x 105 d | TV NOT DETECTED | TV DETECTED |
| Haemophilus ducreyi | 1 x 106 d | TV NOT DETECTED | TV DETECTED |
| Haemophilus influenzae | 3 x 106 | TV NOT DETECTED | TV DETECTED |
| Herpes simplex virus If | 1 x 105 | TV NOT DETECTED | TV DETECTED |
| Herpes simplex virus IIf | 1 x 105 | TV NOT DETECTED | TV DETECTED |
| HIV-1f | 2 x 105 | TV NOT DETECTED | TV DETECTED |
| | Concentration
Testeda | Xpert TV Assay Result | |
| Microorganism | | Cross-Reactivity
(- T. vaginalis) | Competitive
Interference
(+ T. vaginalis) |
| Human papilloma virus 16f | 6 x 105 | TV NOT DETECTED | TV DETECTED |
| Kingella dentrificans | 3 x 106 | TV NOT DETECTED | TV DETECTED |
| Kingella kingae | 7 x 106 | TV NOT DETECTED | TV DETECTED |
| Klebsiella oxytoca | 1 x 106 | TV NOT DETECTED | TV DETECTED |
| Klebsiella pneumoniae | 7 x 106 | TV NOT DETECTED | TV DETECTED |
| Lactobacillus acidophilus | 9 x 106 | TV NOT DETECTED | TV DETECTED |
| Lactobacillus brevis | 7 x 106 | TV NOT DETECTED | TV DETECTED |
| Lactobacillus crispatus | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Lactobacillus jensonii | 9 x 106 | TV NOT DETECTED | TV DETECTED |
| Lactobacillus lactis | 1 x 107 | TV NOT DETECTED | TV DETECTED |
| Lactobacillus vaginalis | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Legionella pneumophila | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Leuconostoc
paramensenteroides | 9 x 106 | TV NOT DETECTED | TV DETECTED |
| Listeria monocytogenes | 8 x 106 | TV NOT DETECTED | TV DETECTED |
| Micrococcus luteus | 3 x 106 | TV NOT DETECTED | TV DETECTED |
| Mobiluncus curtisiib | 3 x 106 | TV NOT DETECTED | TV DETECTED |
| Moraxella lacunata | 1 x 106 | TV NOT DETECTED | TV DETECTED |
| Moraxella osloensis | 1 x 106 | TV NOT DETECTED | TV DETECTED |
| Morganella morganii | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Mycobacterium smegmatis | 1 x 106 | TV NOT DETECTED | TV DETECTED |
| Mycoplasma genitalium | 1 x 106d | TV NOT DETECTED | TV DETECTED |
| Mycoplasma hominis | 1 x 106d | TV NOT DETECTED | TV DETECTED |
| Neisseria cinerea | 1 x 106 | TV NOT DETECTED | TV DETECTED |
| Neisseria dentrificans | 7 x 106 | TV NOT DETECTED | TV DETECTED |
| Neisseria elongata | 8 x 106 | TV NOT DETECTED | TV DETECTED |
| Neisseria flava | 9 x 106 | TV NOT DETECTED | TV DETECTED |
| Neisseria flavescens | 8 x 106 | TV NOT DETECTED | TV DETECTED |
| Neisseria gonorrhoeae | 6 x 106 | TV NOT DETECTED | TV DETECTED |
| Neisseria lactamica | 1 x 106 | TV NOT DETECTED | TV DETECTED |
| Neisseria mucosa | 9 x 106 | TV NOT DETECTED | TV DETECTED |
| Neisseria perflava | 1 x 106 | TV NOT DETECTED | TV DETECTED |
| Neisseria polysaccharea | 4 x 106 | TV NOT DETECTED | TV DETECTED |
| Microorganism | Concentration
Testeda | Xpert TV Assay Result | |
| | | Cross-Reactivity
(- T. vaginalis) | Competitive
Interference
(+ T. vaginalis) |
| Neisseria sicca | 1 x 106 | TV NOT DETECTED | TV DETECTED |
| Neisseria subflava | 9 x 106 | TV NOT DETECTED | TV DETECTED |
| Pantoea agglomerans | 4 x 106 | TV NOT DETECTED | TV DETECTED |
| Paracoccus denitrificans | 6 x 106 | TV NOT DETECTED | TV DETECTED |
| Pentatrichomonis hominisc | 1 x 106 | TV NOT DETECTED | TV DETECTED |
| Peptostreptococcus anaerobiusb | 4 x 106 | TV NOT DETECTED | TV DETECTED |
| Peptostreptococcus productusb | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Plesiomonas shigelloides | 4 x 106 | TV NOT DETECTED | TV DETECTED |
| Prevotella biviab | 1 x 106 | TV NOT DETECTED | TV DETECTED |
| Propionibacterium acnesb | 3 x 106 | TV NOT DETECTED | TV DETECTED |
| Proteus mirabilis | 4 x 106 | TV NOT DETECTED | TV DETECTED |
| Proteus vulgaris | 7 x 106 | TV NOT DETECTED | TV DETECTED |
| Providencia stuartii | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Pseudomonas aeruginosa | 4x 106 | TV NOT DETECTED | TV DETECTED |
| Pseudomonas fluorescens | 5 x 106 | TV NOT DETECTED | TV DETECTED |
| Pseudomonas putida | 4 x 106 | TV NOT DETECTED | TV DETECTED |
| Rahnella aquatilis | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Rhodospirillum rubrum | 1 x 106d | TV NOT DETECTED | TV DETECTED |
| Saccharomyces cerevisiaee | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Salmonella minnesota | 1 x 106 | TV NOT DETECTED | TV DETECTED |
| Salmonella typhimurium | 5 x 106 | TV NOT DETECTED | TV DETECTED |
| Serratia marcescens | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Staphylococcus aureus | 9 x 106 | TV NOT DETECTED | TV DETECTED |
| Staphylococcus epidermidis | 5 x 106 | TV NOT DETECTED | TV DETECTED |
| Staphylococcus saprophyticus | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Streptococcus agalactiae | 6 x 106 | TV NOT DETECTED | TV DETECTED |
| Streptococcus bovis | 4 x 106 | TV NOT DETECTED | TV DETECTED |
| Streptococcus mitis | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Streptococcus mutans | 5 x 106 | TV NOT DETECTED | TV DETECTED |
| Streptococcus pneumoniae | 1 x 106 | TV NOT DETECTED | TV DETECTED |
| Streptococcus pyogenes | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Streptococcus salivarius | 3 x 106 | TV NOT DETECTED | TV DETECTED |
| Microorganism | Concentration
Testeda | Xpert TV Assay Result | |
| | | Cross-Reactivity
(- T. vaginalis) | Competitive
Interference
(+ T. vaginalis) |
| Streptococcus sanguis | 2 x 106 | TV NOT DETECTED | TV DETECTED |
| Streptomyces griseinus | 4 x 106 | TV NOT DETECTED | TV DETECTED |
| Trichomonas tenaxc | 1 x 105 | TV DETECTED | TV DETECTED |
| Trichomonas tenaxc | 1 x 103 | TV DETECTED | TV DETECTED |
| Trichomonas tenaxc | 1 x 102 | TV NOT DETECTED | TV DETECTED |
| Ureaplasma parvum | 1 x 106d | TV NOT DETECTED | TV DETECTED |
| Ureaplasma urealyticum | 1 x 106 | TV NOT DETECTED | TV DETECTED |
| Vibrio parahaemolyticus | 3 x 106 | TV NOT DETECTED | TV DETECTED |
| Yersinia enterocolitica | 3 x 106 | TV NOT DETECTED | TV DETECTED |

16

17

18

19

a. Tests run ≥ 10° CFU/mL for bacteria and fungi, ≥10° genomes/mL for yeast, ≥10° TCIDs0/mL or ≥10° genomes/mL for viruses and ≥10° cells/mL for protozoans.

b. Anaerobic organism

c. Protozoan

d. Genome equivalents tested (DNA)

e. Fungal organism

f. Virus

Additional three microorganisms, Dientamoeba fragilis, Agrobacterium radiobacter, and Erwinia herbicola, were not available for direct testing. An in silico analysis was conducted using the Basic Local Alignment Search Tool (BLAST) to compare the Xpert TV Assay primer and probe sequences with all available sequences associated with these three microorganisms in the GenBank database. Available sequence data for D. fragilis was examined and showed a maximum of 7% homology to the Xpert TV primer and probe sequences. Available sequence data for A. radiobacter was examined and showed a maximum of 38% homology to the Xpert TV primer and probe sequences. Available sequence data for E. herbicola was examined and showed a maximum of 10% homology to the Xpert TV primer and probe sequences. Results are shown in Table 5-6.

Strain	Accession Number	% Homology
Dientamoeba fragilis	KC967121.1	7%
Agrobacterium radiobacter	CP000629.1	38%
Erwinia herbicola	NG_035384.1	10%

Table 5-6. In silico Analytical Specificity Determination for Xpert TV Assay

Analytical Reactivity (Inclusivity)

The analytical inclusivity of the Xpert TV Assay was evaluated by testing 17 T. vaginalis strains diluted in either negative pooled vaginal swab matrix in Cepheid Xpert Swab Transport Reagent or negative pooled urine in Cepheid Xpert Urine Transport Reagent. All T. vaginalis strains were tested in triplicate at a concentration of 3X the

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analytical LoD for the respective specimen type (6 cells/mL for vaginal swabs and 7.5 cells/mL for urine). All strains tested were reported as TV DETECTED. Results are shown in Table 5-7. Positive and negative controls were included in the study. The inclusivity for the 17 T. vaginalis strains tested was 100%.

| Isolate ATCC # | Isolation Source | Results Vaginal
Swab | Results Urine |
|----------------|--------------------------------------|-------------------------|---------------|
| 30001 | Vaginal exudate | TV DETECTED | TV DETECTED |
| 30184 | Vaginal swab | TV DETECTED | TV DETECTED |
| 30187 | Endocervical swab | TV DETECTED | TV DETECTED |
| 30188 | Vagina | TV DETECTED | TV DETECTED |
| 30236 | Endocervical swab | TV DETECTED | TV DETECTED |
| 30240 | Vaginal pool | TV DETECTED | TV DETECTED |
| 30245 | Vaginal and
Endocervical material | TV DETECTED | TV DETECTED |
| 30247 | Vagina | TV DETECTED | TV DETECTED |
| 50138 | human | TV DETECTED | TV DETECTED |
| 50139 | human | TV DETECTED | TV DETECTED |
| 50141 | human | TV DETECTED | TV DETECTED |
| 50143 | human | TV DETECTED | TV DETECTED |
| 50147 | human | TV DETECTED | TV DETECTED |
| 50167 | Vagina | TV DETECTED | TV DETECTED |
| 50183 | Prostatic fluid | TV DETECTED | TV DETECTED |
| PRA-95 | Vaginal exudate | TV DETECTED | TV DETECTED |
| PRA-98 | human | TV DETECTED | TV DETECTED |

Table 5-7: Analytical Reactivity (Inclusivity) of Xpert TV Assav

Interfering Substances Study

The performance of the Xpert TV Assay was evaluated with potentially interfering endogenous and exogenous substances that may be present in the urogenital tract.

All substances were tested in the presence and absence of 3X LoD T. vaginalis (ATCC strain 30001) to determine if there was interference with the Xpert TV Assay. Substances were individually diluted into either pooled Trichomonas vaginalisnegative urine matrix (patient urine added to Cepheid Urine Transport Reagent) or pooled Trichomonas vaginalis-negative vaginal swab matrix (vaginal swabs collected into Cepheid Swab Transport Reagent). Positive and negative controls were included in the study.

For each interfering substance, eight replicates were tested for each set of samples (either T. vaginalis negative or T. vaginalis positive in clinical matrix). Tables 5-8 and 5-9 show the substances that were tested, the test concentrations, and the matrix in which they were diluted. One substance, blood at > 60% v/v demonstrated interference (result of TV NOT DETECTED in the presence of TV) in the vaginal swab matrix samples. Blood was subjected to repeat analysis at various lower concentrations until a result of TV

21

DETECTED was obtained (50% v/v). For the other conditions and substances tested, all TV positive samples remained positive and all TV negative samples remained negative, indicating that there was no interference causing false negative or false positive results with the Xpert TV Assay for these substances.

Class/Substance	Active Ingredient	Concentration Tested
Blood	Blood	0.3% v/v, 1% v/v
Seminal Fluid	Seminal Fluid	5.0% v/v
Mucus	Mucin	0.8% w/v
Analgesics &
Antibiotics	Acetylsalicylic Acid 500mg	40 mg/mL
	Acetaminophen	3.2 mg/mL
	Azithromycin	1.8 mg/mL
	Doxycycline	3.6 mg/mL
OTC Deodorant &
Powders	PEG-20; PEG-32;
PEG-20 Stearate	0.25% w/v
	Nanoxynol-9	0.25% w/v

Albumin	BSA	10 mg/ml
Glucose	Glucose	10 mg/ml
Bilirubin	Bilirubin	1 mg/ml
Acidic Urine (pH 4.0)	Urine + N-Acetyl-L-Cysteine	pH 4.0
Alkaline Urine (pH 9.0)	Urine + Ammonium Citrate	pH 9.0
Leukocytes	Leukocytes	105 cells/mL
Intravaginal Hormones	Progesterone; Estradiol	7 mg/mL Progesterone +
0.07 mg/mL Beta Estradiol

Table 5-8: Potentially Interfering Substances in Urine Samples

Table 5-9: Potentially Interfering Substances in Swab Samples

Class/Substance	Active Ingredient	Concentration Tested
Blooda	Blood	10%, 50%, 60% v/v
Seminal Fluid	Seminal Fluid	5.0% v/v
Mucus	Mucin	0.8% w/v
Over the counter (OTC)
Vaginal Products;
Contraceptives; Vaginal
treatments	Benzocaine 5%;
Resorcinol 2%	0.25% w/v
	Clotrimazole 2%	0.25% w/v
	Miconazole Nitrate 2%	0.25% w/v
	Tioconazole	0.25% w/v
	5% w/w Aciclovir	0.25% w/v
	Glycerin, Propylene glycol	0.25% w/v
	Glycerin; Carbomer	0.12% w/v
	Glycerin, Hydroxyethyl cellulose	0.25% w/v
	Goldenseal 3X HPUS;
Kreosotum 12X HPUS	0.25% w/v
	Povidone-iodine 10%	0.25% v/v
	Nonoxynol-9 12.5%	0.25% w/v

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Class/Substance	Active Ingredient	Concentration Tested
Hemorrhoidal Cream	Glycerin 14%;
Pramoxine HCl 1%	0.25% w/v
Leukocytes	Leukocytes	$10^5$ cells/mL
Intravaginal Hormones	Progesterone; Estradiol	7 mg/mL Progesterone +
0.07 mg/mL Beta Estradiol

a. In tests with substances diluted into pooled T. vaginalis-positive swab matrix, assay interference was observed in tests with blood at 60% v/v. No assay interference was observed in tests with blood at 50% v/v. This is addressed in Limitations in the package insert.

Carry-Over Contamination

This study was conducted to demonstrate that single-use, self-contained GeneXpert cartridges prevent carry-over contamination in negative samples run after very high positive samples in the same GeneXpert module. A negative sample (T. vaginalis negative vaginal swabs in Cepheid Xpert Swab Transport Reagent) was run followed by 20 rounds of high positive sample (T. vaginalis ATCC 30001 at 10° cells/mL diluted in vaginal swab matrix) alternating with a negative sample in two separate GeneXpert modules for a total of 40 high positive and 42 negative samples for each module. This testing scheme resulted in a total of 82 runs (40 positive + 42 negative single-use samples). There was no evidence of carry-over contamination as all 40 positive samples were correctly reported as TV DETECTED and all 42 negative samples were correctly reported as TV NOT DETECTED.

Linearity

Not applicable, the Xpert TV Assay is a qualitative assay.

Clinical Studies

Clinical Performance

Performance characteristics of the Xpert TV Assay were determined in a multi-site prospective investigational study by comparing the results from the Xpert TV Assay to a patient infected status (PIS) algorithm comprised of culture and validated bidirectional sequencing (primary sequencing) for male urine, or an FDA cleared NAAT test and culture for female specimen types.

Study participants included consenting asymptomatic and symptomatic, sexually active males and females seen at locations including, but not limited to: OB/GYN, sexually transmitted disease (STD), and family planning clinics. The average age among eligible female study participants was 33.5 years (range = 18 to 78 years). The average age among eligible male study participants was 36.2 years (range = 16 to 78 years).

The study specimens consisted of prospectively collected male urine, endocervical swabs, and patient-collected vaginal swabs (collected in a clinical setting). Clinician-collected vaginal swabs were collected for testing by the reference NAAT test and culture. Samples were collected from 17 clinical sites and tested at 11 sites. Reference testing was performed at 3 central laboratories.

A study participant was considered to be infected by PIS if either of the two reference

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test results were positive. The subject was considered to be not infected by PIS when both reference test results were negative.

Performance of the Xpert TV Assay was calculated relative to the PIS for each of the three female specimen types (endocervical swabs, patient-collected vaginal swabs and urine) or to the PIS for male urine, respectively.

Specimens with discrepant results between the Xpert TV Assay and the PIS were analyzed by validated bi-directional Sanger sequencing and results are footnoted in Table 5-10 for informational purposes only.

Among the 10,017 tests performed, 190 had initial ERROR, INVALID, or NO RESULT outcomes (1.90%, 95% CI 1.65-2.18). Of those, 167 specimens yielded valid results upon repeat assay (7 specimens were not retested). The overall valid reporting rate of the assay was 99.8% (9994/10,017).

Results of the Xpert TV Assay were compared to the PIS for determination of sensitivity, specificity, and predictive values. Sensitivity and specificity for TV by specimen type and symptom status are presented in Table 5-10.

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Sample Type	Status	Total (n)	Sens	95% CI	Spec	95% CI
ES	Symp	685	100%
(71/71)	94.9%-100%	98.5%
(605/614)	97.2%-99.3%	10.4%	88.8%	100%
	Asymp	1114	98.1%
(104/106)	93.4%-99.8%	99.1%
(999/1008)	98.3%-99.6%	9.5%	92.0%	99.8%
	Overall	1799	98.9%
(175/177)a	96.0%-99.9%	98.9%
(1604/1622)b	98.3%-99.3%	9.8%	90.7%	99.9%
	Difference	P-Value	P=0.517	-0.70%, 4.48%	P=0.331	-1.69%, 0.54%
	Symp	682	98.6%
(73/74)	92.7%-100%	99.5%
(605/608)	98.6%-99.9%	10.9%	96.1%	99.8%
PC-VS	Asymp	1109	95.0%
(113/119)	89.3%-98.1%	99.6%
(986/990)	99.0%-99.9%	10.7%	96.6%	99.4%
	Overall	1791	96.4%
(186/193)c	92.7%-98.5%	99.6%
(1591/1598)d	99.1%-99.8%	10.8%	96.4%	99.6%
	Difference	P-Value	P=0.254	-1.04%, 8.42%	P=1.000	-0.77%, 0.59%
	Symp	688	98.6%
(71/72)	92.5%-100%	99.8%
(615/616)	99.1%-100%	10.5%	98.6%	99.8%
UR-F	Asymp	1105	98.2%
(109/111)	93.6%-99.8%	99.6%
(990/994)	99.0%-99.9%	10.0%	96.5%	99.8%
	Overall	1793	98.4%
(180/183)e	95.3%-99.7%	99.7%
(1605/1610)f	99.3%-99.9%	10.2%	97.3%	99.8%
	Difference	P-Value	P=1.000	-3.25%, 4.08%	P=0.655	-0.27%, 0.75%
	Symp	1088	87.5%
(28/32)	71.9%-95.0%	99.8%
(1054/1056)	99.3%-99.9%	2.9%	93.3%	99.6%
UR-M	Asymp	3523	90.3%
(84/93)	82.6%-94.8%	99.2%
(3401/3430)	98.8%-99.4%	2.6%	74.3%	99.7%
	Overall	4611	89.6%
(112/125)g	83.0%-93.8%	99.3%
(4455/4486)h	99.0%-99.5%	2.7%	78.3%	99.7%
	Difference		P=0.738	-15.8%, 10.1%	P=0.020	0.25%, 1.06%

Table 5-10: Xpert TV vs. PIS by Symptomatic Status

TP=tne positive, PP=false positive, TN=false negative, ES=ndocervical swab, PC-VS=patient-ollected vaginal swab, UR-M=rale urne a. Testing results by sequencing: 1 of 2 FN was TV positive; 1 of 2 was TV negative.

b. Testing results by sequencing: 8 of 18 FP were TV positive; 10 of 18 were TV negative.

c. Testing results by sequencing: 3 of 7 FN were TV positive; 4 of 7 were TV negative.

d. Testing results by sequencing: 5 of 7 FP were TV positive; 2 of 7 were TV negative.

e. Testing results by sequencing: 3 of 3 FN were TV negative.

f. Testing results by sequencing: 5 of 5 FP were TV negative.

g. Testing results by secondary sequencing: 9 of 13 false negative; 4 of 13 were TV positive.

h. Testing results by secondary sequencing: 27 of 31 false positive; 4 of 31 were TV negative.

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Cycle Threshold (Ct) Frequency Distribution

Patient-collected vaginal swabs, endocervical swabs and urine specimens were collected from 1867 females and urine specimens were collected from 4626 males at 17 collection sites in the US. The frequency distribution of Xpert TV Assay positive results for the 197 Trichomonas vaginalis infected female study subjects and 125 Trichomonas vaginalis infected male study subjects are shown in Figure 5-1.

Image /page/25/Figure/2 description: The image is a histogram displaying the distribution of Ct values. The x-axis represents Ct values, ranging from 10 to 40, while the y-axis represents the number of specimens, ranging from 0 to 60. The histogram shows a distribution that peaks between Ct values of 25 and 30, indicating that most specimens have Ct values within this range. The number of specimens decreases as Ct values move away from this peak in either direction.

Figure 5-1: Ct Distribution of Patients Designated as Positive for TV Based on PIS Algorithm

Reproducibility Study

Intra-site reproducibility of the Xpert TV Assay was evaluated at three sites (two external, one in-house). Site 1 used an Infinity-80 instrument. Sites 2 and 3 used GeneXpert Dx instruments. Specimens were created by spiking Trichomonas vaginalis (ATCC® 30001™) into pooled, Trichomonas vaginalis negative urine (patient urine added to Cepheid Urine Transport Reagent) or vaginal swab matrix (vaginal swabs collected into Cepheid Swab Transport Reagent). The specimens were prepared at concentration levels representing high negative (below LoD), LoD (~1X LoD), moderate positive (~3X LoD), and negative (Trichomonas vaginalis negative clinical matrix). A panel of 8 specimens (4 in urine and 4 in vaginal swab matrix) was tested twice per day. on 12 different days, by two different operators, at each of three sites (8 specimens x 2 replicates x 12 days x 2 operators x 3 sites = 1,152 observations total). Three lots of Xpert TV Assay cartridges were used at each of the 3 testing sites, with each lot used for 4 days of testing. Positive and negative controls were included in the study. The Xpert TV Assay was performed according to the Xpert TV Assay procedure. The rate of agreement with expected results is shown by site in Table 5-11.

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| Samplea | Site 1
(Infinity-80) | | Site 2
(GeneXpert Dx) | | Site 3
(GeneXpert Dx) | | Total
Agreement
by Sample | | | |
|---------------------------------------------|-------------------------|------------------|--------------------------|------------------|--------------------------|------------------|---------------------------------|------------------|------------------|--------------------|
| | Op 1 | Op 2 | Site | Op 1 | Op 2 | Site | Op 1 | Op 2 | Site | |
| FS-Neg | 100%
(24/24) | 100%
(24/24) | 100%
(48/48) | 100%
(24/24) | 100%
(24/24) | 100%
(48/48) | 100%
(24/24) | 100%
(24/24) | 100%
(48/48) | 100%
(144/144) |
| FS-Mod Pos
(~3X LoD;
~6 cells/mL) | 100%
(24/24) | 100%
(24/24) | 100%
(48/48) | 100%
(24/24) | 100%
(24/24) | 100%
(48/48) | 100%
(24/24) | 100%
(24/24) | 100%
(48/48) | 100%
(144/144) |
| FS-LoD
(~1X LoD;
~2 cells/mL) | 95.8%
(23/24) | 100%
(24/24) | 97.9%
(47/48) | 87.5%
(21/24) | 95.8%
(23/24) | 91.7%
(44/48) | 100%
(24/24) | 95.8%
(23/24) | 97.9%
(47/48) | 95.8%
(138/144) |
| FS-High Neg
(below LoD;