cobas TV/MG for use on cobas 6800/8800 systems, cobas TV/MG Positive Control Kit, cobas Buffer Negative Control Kit

K190433 · Roche Molecular Systems, Inc. · QEP · May 22, 2019 · Microbiology

Device Facts

• Record ID: K190433
• Device Name: cobas TV/MG for use on cobas 6800/8800 systems, cobas TV/MG Positive Control Kit, cobas Buffer Negative Control Kit
• Applicant: Roche Molecular Systems, Inc.
• Product Code: QEP · Microbiology
• Decision Date: May 22, 2019
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3393
• Device Class: Class 2

Intended Use

cobas TV/MG on the cobas 6800/8800 Systems is an automated, qualitative in vitro nucleic acid diagnostic test that utilizes real-time polymerase chain reaction (PCR), for the direct detection of Trichomonas vaginalis (TV) and Mycoplasma genitalium (MG) DNA in male or female urine, self-collected vaginal swab specimens (collected in a clinical setting), clinician-collected vaginal swab specimens, and endocervical specimens, all collected in cobas PCR Media (Roche Molecular Systems, Inc.). cobas TV/MG also detects TV DNA in cervical specimens collected in PreservCyt solution and MG DNA in self-collected meatal swab specimens (collected in a cliniciancollected meatal swab specimens. This test is intended as an aid in the diagnosis of TV and MG infections in individuals suspected to have TV or MG infection.

Device Story

Automated in vitro diagnostic test for T. vaginalis (TV) and M. genitalium (MG) DNA detection; utilizes real-time PCR on cobas 6800/8800 systems. Input: urine, vaginal/endocervical/meatal swabs, or cervical specimens in transport media. Process: automated nucleic acid extraction/purification using magnetic glass particles; PCR amplification with target-specific primers and fluorescent probes (TaqMan); AmpErase enzyme prevents amplicon contamination. Output: qualitative positive/negative results for TV and MG. Used in clinical laboratories; operated by trained personnel. Results reviewed on system screen or report; aids diagnosis of TV/MG infections. Benefits: rapid, automated, high-throughput detection of STIs.

Clinical Evidence

Multi-site prospective clinical study (n=2,194 subjects). Performance compared to Patient Infected Status (PIS) composite reference (FDA-cleared NAATs, culture, and lab-developed NAATs). TV sensitivity 94.7-100% and specificity 96.8-99.2% across specimen types. MG sensitivity 78.9-100% and specificity 96.3-99.3%. Invalid rate 0.18%.

Technological Characteristics

Automated nucleic acid extraction/purification using magnetic glass particles. Real-time PCR amplification with target-specific primers and fluorescent reporter/quencher probes (TaqMan). Master mix includes dUTP and AmpErase for carryover prevention. Analyzed on cobas 6800/8800 systems. Software-based result interpretation.

Indications for Use

Indicated for detection of T. vaginalis and M. genitalium DNA in male/female urine, vaginal swabs (self/clinician-collected), endocervical specimens, cervical specimens (TV only), and meatal swabs (MG only). Intended for individuals suspected of infection.

Regulatory Classification

Identification

A device to detect nucleic acids from non-viral microorganism(s) causing sexually transmitted infections and associated resistance marker(s) is an in vitro diagnostic device intended for the detection and identification of nucleic acids from non-viral microorganism(s) and their associated resistance markers in clinical specimens collected from patients suspected of sexually transmitted infections. The device is intended to aid in the diagnosis of non-viral sexually transmitted infections in conjunction with other clinical and laboratory data. These devices do not provide confirmation of antibiotic susceptibility since mechanisms of resistance may exist that are not detected by the device.

Special Controls

A device to detect nucleic acids from non-viral microorganism(s) causing sexually transmitted infections and associated resistance marker(s) must comply with the following special controls: (1) The intended use for the 21 CFR 809.10 labeling must include a detailed description of targets the device detects, the results provided to the user, the clinical indications appropriate for test use, and the specific population(s) for which the device is intended. (2) Any sample collection device used must be FDA-cleared, -approved, or -classified as 510(k) exempt (standalone or as part of a test system) for the collection of specimen types claimed by this device: alternatively, the sample collection device must be cleared in a premarket submission as a part of this device. (3) The 21 CFR 809.10(b) labeling must include: (i) A detailed device description, including reagents, instruments, ancillary materials, all control elements, and a detailed explanation of the methodology, including all pre-analytical methods for processing of specimens; (ii) Detailed discussion of the performance characteristics of the device for all claimed specimen types based on analytical studies, including, but not limited to. Limit of Detection, inclusivity, cross-reactivity, interfering substances, competitive inhibition, carryover/cross contamination, specimen stability, with-in lab precision, and reproducibility, as appropriate; (iii) Detailed descriptions of the test procedure, the interpretation of test results for clinical specimens, and acceptance criteria for any quality control testing. (iv) Limiting statements indicating that: (A)a negative test result does not preclude the possibility of infection; (B) the test results should be interpreted in conjunction with other clinical and laboratory data available to the clinician; (C) reliable results are dependent on adequate specimen collection, transport, storage, and processing. Failure to observe procedures in any one of these steps can lead to incorrect results; and (D)if appropriate (e.g., recommended by CDC, by current well-accepted clinical guidelines, or by published peer reviewed research), that the clinical performance is inferior in a specific clinical subpopulation or for a specific claimed specimen type. (v) If the device is intended to detect antimicrobial resistance markers, limiting statements, as appropriate, indicating that: (A)negative results for claimed resistance markers do not indicate susceptibility of detected microorganisms, as resistance markers not measured by the assay or other potential mechanisms of antibiotic resistance may be present; (B) detection of resistance markers cannot be definitively linked to specific microorganisms and the source of a detected resistance marker may be an organism not detected by the assay, including colonizing flora; (C) detection of antibiotic resistance markers may not correlate with phenotypic gene expression; and (D) therapeutic failure or success cannot be determined based on the assay results, since nucleic acid may persist following appropriate antimicrobial therapy. (4) Design verification and validation must include: (i) Detailed device description documentation, including, but not limited to, methodology from obtaining sample to result, design of primer/probe sequences, rationale for target sequence selection, and computational path from collected raw data to reported result (e.g., how collected raw signals are converted into a reported result). (ii) Detailed documentation of analytical studies including but not limited to, Limit of Detection, inclusivity, cross-reactivity, microbial interference, interfering substances, competitive inhibition, carryover/cross contamination, specimen stability, with-in lab precision, and reproducibility, as appropriate. (iii) Detailed documentation and performance results from a clinical study that includes prospective (sequential) samples for each claimed specimen type and, when determined to be appropriate by FDA, additional characterized clinical samples. The study must be performed on a study population consistent with the intended use population and compare the device performance to results obtained from FDA accepted comparator methods. Documentation from the clinical studies must include the clinical study protocol (including a predefined statistical analysis plan) study report, testing results, and results of all statistical analyses. (iv) A detailed description of the impact of any software, including, but not limited to, software applications and hardware-based devices that incorporate software, on the device's functions.

*Classification.* Class II (special controls). The special controls for this device are:(1) The intended use for the labeling required under § 809.10 of this chapter must include a detailed description of targets the device detects, the results provided to the user, the clinical indications appropriate for test use, and the specific population(s) for which the device is intended. (2) Any sample collection device used must be FDA-cleared, -approved, or -classified as 510(k) exempt (standalone or as part of a test system) for the collection of specimen types claimed by this device; alternatively, the sample collection device must be cleared in a premarket submission as a part of this device. (3) The labeling required under § 809.10(b) of this chapter must include: (i) A detailed device description, including reagents, instruments, ancillary materials, all control elements, and a detailed explanation of the methodology, including all pre-analytical methods for processing of specimens; (ii) Detailed discussion of the performance characteristics of the device for all claimed specimen types based on analytical studies, including Limit of Detection, inclusivity, cross-reactivity, interfering substances, competitive inhibition, carryover/cross contamination, specimen stability, within lab precision, and reproducibility, as appropriate; (iii) Detailed descriptions of the test procedure, the interpretation of test results for clinical specimens, and acceptance criteria for any quality control testing; (iv) Limiting statements indicating that: (A) A negative test result does not preclude the possibility of infection; (B) The test results should be interpreted in conjunction with other clinical and laboratory data available to the clinician; (C) Reliable results are dependent on adequate specimen collection, transport, storage, and processing. Failure to observe proper procedures in any one of these steps can lead to incorrect results; and (D) If appropriate ( *e.g.,* recommended by the Centers for Disease Control and Prevention, by current well-accepted clinical guidelines, or by published peer reviewed research), that the clinical performance is inferior in a specific clinical subpopulation or for a specific claimed specimen type; and(v) If the device is intended to detect antimicrobial resistance markers, limiting statements, as appropriate, indicating that: (A) Negative results for claimed resistance markers do not indicate susceptibility of detected microorganisms, as resistance markers not measured by the assay or other potential mechanisms of antibiotic resistance may be present; (B) Detection of resistance markers cannot be definitively linked to specific microorganisms and the source of a detected resistance marker may be an organism not detected by the assay, including colonizing flora; (C) Detection of antibiotic resistance markers may not correlate with phenotypic gene expression; and (D) Therapeutic failure or success cannot be determined based on the assay results, since nucleic acid may persist following appropriate antimicrobial therapy. (4) Design verification and validation must include: (i) Detailed device description documentation, including methodology from obtaining sample to result, design of primer/probe sequences, rationale for target sequence selection, and computational path from collected raw data to reported result ( *e.g.,* how collected raw signals are converted into a reported result).(ii) Detailed documentation of analytical studies, including, Limit of Detection, inclusivity, cross-reactivity, microbial interference, interfering substances, competitive inhibition, carryover/cross contamination, specimen stability, within lab precision, and reproducibility, as appropriate. (iii) Detailed documentation and performance results from a clinical study that includes prospective (sequential) samples for each claimed specimen type and, when determined to be appropriate by FDA, additional characterized clinical samples. The study must be performed on a study population consistent with the intended use population and compare the device performance to results obtained from FDA accepted comparator methods. Documentation from the clinical studies must include the clinical study protocol (including a predefined statistical analysis plan) study report, testing results, and results of all statistical analyses. (iv) A detailed description of the impact of any software, including software applications and hardware-based devices that incorporate software, on the device's functions.

Predicate Devices

• Aptima Mycoplasma genitalium assay (DEN180047)

Submission Summary (Full Text)

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Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

Roche Molecular Systems, Inc. Nobuko Nakajima Director, Regulatory Affairs 4300 Hacienda Drive Pleasanton, California 94588-2722 May 22, 2019

### Re: K190433

| Trade/Device Name: | cobas TV/MG for use on cobas 6800/8800 systems, cobas TV/MG Positive<br>Control Kit, cobas Buffer Negative Control Kit |
|--------------------|-----------------------------------------------------------------------------------------------------------------------------------------------|
| Regulation Number: | 21 CFR 866.3393 |
| Regulation Name: | Device to Detect Nucleic Acids from Non-Viral Microorganism(s) Causing<br>Sexually Transmitted Infections and Associated Resistance Marker(s) |
| Regulatory Class: | Class II |
| Product Code: | QEP |
| Dated: | February 21, 2019 |
| Received: | February 22, 2019 |

Dear Nobuko Nakajima:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

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

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal

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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 Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.htm); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

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

For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (http://www.fda.gov/DICE) for more information or contact DICE by email ([e-mail]) or phone [phone] or 301-796-7100).

Sincerely,

for

Uwe Scherf, Ph.D. Director Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health

Enclosure

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### Indications for Use

510(k) Number (if known) K190433

### Device Name

cobas TV/MG for use on cobas 6800/8800 systems, cobas TV/MG Positive Control Kit, cobas Buffer Negative Control Kit

### Indications for Use (Describe)

cobas TV/MG on the cobas 6800/8800 Systems is an automated, qualitative in vitro nucleic acid diagnostic test that utilizes real-time polymerase chain reaction (PCR), for the direct detection of Trichomonas vaginalis (TV) and Mycoplasma genitalium (MG) DNA in male or female urine, self-collected vaginal swab specimens (collected in a clinical setting), clinician-collected vaginal swab specimens, and endocervical specimens, all collected in cobas PCR Media (Roche Molecular Systems, Inc.). cobas TV/MG also detects TV DNA in cervical specimens collected in PreservCyt solution and MG DNA in self-collected meatal swab specimens (collected in a cliniciancollected meatal swab specimens. This test is intended as an aid in the diagnosis of TV and MG infections in individuals suspected to have TV or MG infection.

A vaginal swab (self-collected or clinician-collected) is the preferred specimen type for MG testing in females due to higher sensitivity compared to endocervical swabs and urine. For males, urine is the preferred specimen type due to higher sensitivity compared to meatal swabs. If vaginal swab or male urine is not used and MG testing, is negative, further testing with the preferred specimen type may be indicated if M. genitalium infection is strongly suspected.

### Ancillary Collection Kits:

The cobas PCR Media Dual Swab Sample Kit is used to collect and transport human specimens. The cobas PCR Media serves as a nucleic acid stabilizing transport and storage medium for human specimens.

- Note: This kit has been validated for use with the following tests:
- · cobas CT/NG v2.0 Test (for use on the cobas 4800 Systems)
- cobas CT/NG for use on cobas 6800/8800 Systems
- · cobas TV/MG for use on the cobas 6800/8800 Systems

The cobas PCR Media Uni Swab Sample Kit is used to collect and transport human specimens.

The cobas PCR Media serves as a nucleic acid stabilizing transport and storage medium for human specimens.

Note: This kit has been validated for use with the following tests:

· cobas CT/NG v2.0 Test (for use on the cobas 4800 Systems)

- cobas CT/NG for use on cobas 6800/8800 Systems
- · cobas TV/MG for use on the cobas 6800/8800 Systems
- cobas Cdiff Test for use on the cobas 4800 System
- cobas Cdiff for use on the cobas Liat System

The cobas PCR Urine Sample Kit is used to collect and transport urine specimens. The cobas PCR Media serves as a nucleic acid stabilizing transport and storage medium for urine specimens.

Note: This kit has been validated for use with the following tests:

- · cobas CT/NG v2.0 Test (for use on cobas 4800 Systems)
- cobas CT/NG for use on cobas 6800/8800 Systems
- cobas TV/MG for use on cobas 6800/8800 Systems

Type of Use (Select one or both, as applicable)

X Prescription Use (Part 21 CFR 801 Subpart D)

Over-The-Counter Use (21 CFR 801 Subpart C)

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# cobas® TV/MG 510(k) Summary

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of 21 CFR 807.92.

| Submitter Name | Roche Molecular Systems, Inc. |
|----------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Address | 4300 Hacienda Drive<br>Pleasanton, CA, 94588-2722 |
| Contact | Nobuko Nakajima<br>Phone: [phone]<br>FAX: [phone]<br>Email: [e-mail] |
| Date Prepared | February 21, 2019 |
| Proprietary Name | cobas® TV/MG<br>for use on cobas® 6800/8800 systems |
| Classification Name | 21 CFR 866.3393 Nucleic acid detection system for non-viral microorganism(s)<br>causing sexually transmitted infections<br>21 CFR 866.3860: Trichomonas vaginalis nucleic acid amplification test system |
| Product Codes | QEP: 21 CFR 866.3393<br>OUY: 21 CFR 866.3860 |
| Predicate Devices | Aptima Mycoplasma genitalium assay (DEN180047) |
| Establishment Registration | Roche Molecular Systems, Inc. (2243471) |

### 1. DEVICE DESCRIPTION

cobas® TV/MG on the cobas® 6800/8800 Systems is an automated, qualitative in vitro nucleic acid diagnostic test for the direct detection of Trichomonas vaginalis (TV) and Mycoplasma genitalium (MG) DNA in male or female urine, self-collected vaginal swab specimens (collected in a clinical setting), clinician-collected vaginal swab specimens, and endocervical specimens collected in cobas® PCR Media (Roche Molecular Systems, Inc.). cobas® TV/MG also detects TV DNA in cervical specimens collected in PreservCyt® Solution and MG DNA in self-collected meatal swab specimens (collected in a clinical setting) and clinician-collected meatal swab specimens. The DNA Internal Control, used to monitor the entire sample preparation and PCR

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amplification process, is introduced into each specimen during sample processing. In addition, the test utilizes external controls (low titer positive control and a negative control).

### Principles of the procedure 1.1.

cobas® TV/MG is based on fully automated sample preparation (nucleic acid extraction and purification) followed by PCR amplification and detection. The cobas® 6800/8800 Systems consist of the sample supply module, the transfer module, the processing module, and the analytic module. Automated data management is performed by the cobas® 6800/8800 software which assigns test results for all tests as positive, negative or invalid. Results can be reviewed directly on the system screen, exported, or printed as a report.

Nucleic acid from patient samples and added internal control DNA (DNA-IC) molecules are simultaneously extracted. In summary, nucleic acid is released by addition of proteinase and lysis reagent to the sample. The released nucleic acid binds to the silica surface of the added magnetic glass particles. Unbound substances and impurities, such as denatured protein, cellular debris and potential PCR inhibitors are removed with subsequent wash steps and purified nucleic acid is eluted from the magnetic glass particles with elution buffer at elevated temperature. External controls (positive and negative) are processed in the same way with each cobas® TV/MG run.

Selective amplification of target nucleic acid from the sample is achieved by the use of targetspecific forward and reverse primers for TV and MG which are selected from highly-conserved regions within the respective target organism. TV is detected by one selective set of primers and a probe, while MG is detected by using two sets targeting separate regions (dual-target). Selective amplification of DNA IC is achieved by the use of sequence-specific forward and reverse primers which are selected to have no homology with either the TV or MG target regions. A thermostable DNA polymerase enzyme is used for PCR amplification. The target and DNA-IC sequences are amplified simultaneously utilizing a universal PCR amplification profile with predefined temperature steps and number of cycles. The master mix includes deoxyuridine triphosphate (dUTP), instead of deoxythymidine triphosphate (dTTP), which is incorporated into the newly synthesized DNA (amplicon). Any contaminating amplicon from previous PCR runs is eliminated by the AmpErase enzyme, which is included in the PCR master mix, during the first thermal cycling step. However, newly formed amplicons are not eliminated since the AmpErase enzyme is inactivated once exposed to temperatures above 55°C.

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The cobas® TV/MG master mix contains one detection probe specific for the TV target sequence, two detection probes specific for the MG target sequences and one for the DNA-IC. The probes are labeled with target specific fluorescent reporter dyes allowing simultaneous detection of TV target, MG targets and DNA-IC in three different target channels. When not bound to the target sequence, the fluorescent signal of the intact probes is suppressed by a quencher dye. During the PCR amplification step, hybridization of the probes to the specific single-stranded DNA template results in cleavage of the 5' to 3' exonuclease activity of the DNA polymerase resulting in separation of the reporter and quencher dyes and the generation of a fluorescent signal. With each PCR cycle, increasing amounts of cleaved probes are generated and the cumulative signal of the reporter dye increases concomitantly. Real-time detection and discrimination of PCR products is accomplished by measuring the fluorescence of the released reporter dyes for the TV and MG targets and DNA-IC, respectively.

### Figure 1: cobas® TV/MG for use on the cobas® 6800/8800 Systems

Image /page/6/Figure/2 description: The image shows three boxes of Cobas control kits. The first box is the Cobas TV/MG Positive Control Kit, which is a positive control kit for use on the Cobas 6800/8800 Systems. The second box is the Cobas TV/MG Qualitative nucleic acid test for use on the Cobas 6800/8800 Systems. The third box is the Cobas Buffer Negative Control Kit, which is a buffer negative control kit for use on the Cobas 6800/8800 Systems.

#### 2. INDICATIONS FOR USE

cobas TV/MG on the cobas 6800/8800 Systems is an automated, qualitative in vitro nucleic acid diagnostic test that utilizes real-time polymerase chain reaction (PCR), for the direct detection of Trichomonas vaginalis (TV) and Mycoplasma genitalium (MG) DNA in male or female urine, self-collected vaginal swab specimens (collected in a clinical setting), clinician-collected vaginal swab specimens, and endocervical specimens, all collected in cobas PCR Media (Roche Molecular Systems, Inc.). cobas TV/MG also detects TV DNA in cervical specimens collected in PreservCyt solution and MG DNA in self-collected meatal swab specimens (collected in a clinical setting) and clinician-collected meatal swab specimens. This test is intended as an aid in the diagnosis of TV and MG infections in individuals suspected to have TV or MG infection.

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A vaginal swab (self-collected or clinician-collected) is the preferred specimen type for MG testing in females due to higher sensitivity compared to endocervical swabs and urine. For males, urine is the preferred specimen type due to higher sensitivity compared to meatal swabs. If vaginal swab or male urine is not used and MG testing is negative, further testing with the preferred specimen type may be indicated if M. genitalium infection is strongly suspected.

### Ancillary Collection Kits 2.1.

The cobas® PCR Media Dual Swab Sample Kit is used to collect and transport human specimens. The cobas® PCR Media serves as a nucleic acid stabilizing transport and storage medium for human specimens.

Note: This kit has been validated for use with the following tests:

- cobas® CT/NG v2.0 Test (for use on the cobas® 4800 Systems) .
- cobas® CT/NG for use on cobas® 6800/8800 Systems .
- cobas® TV/MG for use on the cobas® 6800/8800 Systems .

The cobas® PCR Media Uni Swab Sample Kit is used to collect and transport human specimens.

The cobas® PCR Media serves as a nucleic acid stabilizing transport and storage medium for human specimens.

Note: This kit has been validated for use with the following tests:

- cobas® CT/NG v2.0 Test (for use on the cobas® 4800 Systems) .
- cobas® CT/NG for use on cobas® 6800/8800 Systems .
- cobas® TV/MG for use on the cobas® 6800/8800 Systems .
- cobas® Cdiff Test for use on the cobas® 4800 System .
- cobas® Cdiff for use on the cobas® Liat® System .

The cobas® PCR Urine Sample Kit is used to collect and transport urine specimens. The cobas® PCR Media serves as a nucleic acid stabilizing transport and storage medium for urine specimens.

Note: This kit has been validated for use with the following tests:

- cobas® CT/NG v2.0 Test (for use on cobas® 4800 Systems) .
- cobas® CT/NG for use on cobas® 6800/8800 Systems .

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- · cobas® TV/MG for use on cobas® 6800/8800 Systems
### 3. TECHNOLOGICAL CHARACTERISTICS

The primary technological characteristics and intended use of the RMS cobas® TV/MG for use on the cobas® 6800/8800 systems are substantially equivalent to other legally marketed nucleic acid amplification tests intended for the qualitative detection of Trichomonas vaginalis (TV) and/or Mycoplasma genitalium (MG).

As indicated in Table 1, the cobas® TV/MNG for use on the cobas® 6800/8800 systems is substantially equivalent to significant characteristics of the identified predicate device, Hologic Aptima M. Genitalium Assay (DEN 180047).

| | Predicate Device:<br>Aptima Mycoplasma genitalium Assay<br>(DEN180047) | Submitted Device<br>cobas® TV/MG for use on the<br>cobas® 6800/8800 systems |
|------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended<br>Use | The Aptima Mycoplasma genitalium assay is<br>an in vitro nucleic acid amplification test<br>(NAAT) for the qualitative detection of<br>ribosomal RNA (rRNA) from Mycoplasma<br>genitalium on the fully automated Panther®<br>system. It is intended for use as an aid in the<br>diagnosis of M. genitalium urogenital<br>infections in male and female patients<br>suspected of M. genitalium infection. The<br>assay may be used to test the following<br>specimens: clinician-collected and self-<br>collected vaginal swabs (in a clinical setting),<br>clinician-collected endocervical swabs,<br>female and male urine, clinician-collected<br>male urethral swabs, and self-collected<br>penile meatal swabs (in a<br>clinical setting). For females, a vaginal swab<br>is the preferred specimen type due to higher<br>clinical sensitivity for detecting M. genitalium<br>than other specimen types; however, female<br>urine or clinician-collected endocervical<br>swabs may be used as alternative<br>specimens when vaginal swab specimens<br>are not available. If female urine or clinician-<br>collected endocervical swab specimens test<br>negative, testing with a vaginal swab may be | cobas TV/MG on the cobas 6800/8800<br>Systems is an automated, qualitative in vitro<br>nucleic acid diagnostic test that utilizes real-<br>time polymerase chain reaction (PCR), for the<br>direct detection of Trichomonas vaginalis (TV)<br>and Mycoplasma genitalium (MG) DNA in male<br>or female urine, self-collected vaginal swab<br>specimens (collected in a clinical setting),<br>clinician-collected vaginal swab specimens,<br>and endocervical specimens, all collected in<br>cobas PCR Media (Roche Molecular Systems,<br>Inc.). cobas TV/MG also detects TV DNA in<br>cervical specimens collected in PreservCyt<br>solution and MG DNA in self-collected meatal<br>swab specimens (collected in a clinical setting)<br>and clinician-collected meatal swab<br>specimens. This test is intended as an aid in<br>the diagnosis of TV and MG infections in<br>individuals suspected to have TV or MG<br>infection.<br>A vaginal swab (self-collected or clinician-<br>collected) is the preferred specimen type for<br>MG testing in females due to higher sensitivity<br>compared to endocervical swabs and urine.<br>For males, urine is the preferred specimen<br>type due to higher sensitivity compared to<br>meatal swabs. If vaginal swab or male urine is |
| | Predicate Device:<br>Aptima Mycoplasma genitalium Assay<br>(DEN180047) | Submitted Device<br>cobas® TV/MG for use on the<br>cobas® 6800/8800 systems |
| | indicated, if M. genitalium infection is<br>suspected. | not used and MG testing is negative, further<br>testing with the preferred specimen type may<br>be indicated if M. genitalium infection is<br>strongly suspected. |
| Sample<br>Types | Clinician-collected and self-<br>collected vaginal swabs (in a<br>clinical setting) Clinician-collected endocervical<br>swabs Female and male urine Clinician-collected male urethral<br>swabs, and Self-collected penile meatal swabs<br>(in a clinical setting) | TV and MG:<br>Male and female urine Self-collected/clinician-collected vaginal<br>swab specimens in cobas® PCR Media Endocervical swab specimens in<br>cobas® PCR Media TV only: Cervical specimens in PreservCyt®<br>solution MG only: Self-collected and clinician-collected<br>penile meatal swabs |
| Conditions<br>for use | For prescription use | Same |
| Amplification<br>Technology | Target Capture (TC), Transcription Mediated<br>Amplification (TMA) | Real-time PCR |
| Detection<br>Chemistry | Hybridization Protection Assay (HPA) | Paired reporter and quencher fluorescence<br>labeled probes (TaqMan Technology) using<br>fluorescence resonance energy transfer<br>(FRET) |
| Result<br>Analysis | Final assay results are interpreted<br>based on the analyte signal-to-cutoff (S/CO). | Based on PCR cycle threshold analysis |
| Analyzer | Assay performed on the PANTHER<br>Instrument | cobas® 6800/8800 systems |
| Sample<br>Preparation<br>Procedure | Automated | Same |

### Comparison of the cobas® TV/MG for use on the cobas® 6800/8800 systems with Table 1: the Predicate Devices

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### NON-CLINICAL PERFORMANCE EVALUATION 4.

### 4.1. Analytical sensitivity (Limit of Detection)

The limit of detection of cobas® TV/MG was determined by analysis of serial dilutions of quantified cultures of two T. vaginalis (RP - metronidazole susceptible and CDC085 metronidazole resistant) and two M. genitalium (MG37 and M30) strains. Panels of six to seven

{10}------------------------------------------------

concentration levels with 60-72 replicates per specimen type and strain plus a blank were tested over three unique lots of cobas® TV/MG test reagents, multiple runs, days, operators, and instruments. LoD for each specimen type is shown in Table 2 as the target concentration which can be detected in ≥ 95% of the replicates for all lots.

| Specimen<br>Types | <i>T. vaginalis</i> | | | | <i>M. genitalium</i> | | | |
|------------------------------------------------------------------|---------------------|---------------------|-------------------|---------------------|----------------------|---------------------|--------------------|---------------------|
| | RP strain | | CDC085 strain | | MG37 strain | | M30 strain | |
| | LoD<br>(cells/mL) | Mean<br>Ct<br>Value | LoD<br>(cells/mL) | Mean<br>Ct<br>Value | LoD<br>(copies/mL) | Mean<br>Ct<br>Value | LoD<br>(copies/mL) | Mean<br>Ct<br>Value |
| Endocervical<br>Swab in<br>cobas® PCR<br>Media | 0.2 | 36.3 | 0.2 | 35.6 | 2 | 35.3 | 2 | 36.5 |
| Vaginal Swab<br>in cobas® PCR<br>Media | 0.3 | 35.5 | 0.075 | 36.3 | 4 | 34.5 | 4 | 35.3 |
| Urine in<br>cobas® PCR<br>Media | 0.1 | 35.7 | 0.03 | 35.6 | 0.5 | 35.6 | 1 | 35.8 |
| Meatal Swab<br>in cobas® PCR<br>Media | N/A | N/A | N/A | N/A | 0.5 | 36.0 | 0.5 | 36.6 |
| Cervical<br>Samples<br>collected into<br>PreservCyt®<br>Solution | 0.1 | 36.8 | 0.05 | 36.6 | N/A | N/A | N/A | N/A |

Table 2: Analytical sensitivity (Limit of Detection)

*N/A = Not Applicable

#### Inclusivity 4.2.

Inclusivity of cobas® TV/MG was evaluated by testing eight TV and five MG laboratory strains using one lot of reagents. Testing was performed using TV and MG cultures diluted into contrived negative matrix. Results are shown in Table 4 for TV and MG strains, respectively. Twenty-four replicates per dilution level were tested for each strain in each matrix.

LoD verification TV strains Table 3:

| Strain | Swabs* | | Urine Specimens | | PreservCyt® Specimens | |
|---------|----------|-------|-----------------|-------|-----------------------|-------|
| | cells/mL | % Pos | cells /mL | % Pos | cells /mL | % Pos |
| C-1:NIH | 0.24 | 100 | 0.07 | 100 | 0.11 | 100 |

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| Strain | Swabs* | Urine Specimens | PreservCyt® Specimens | | | |
|--------------|----------|-----------------|-----------------------|-------|-----------|-------|
| | cells/mL | % Pos | cells /mL | % Pos | cells /mL | % Pos |
| 123414 | 0.24 | 100 | 0.07 | 100 | 0.11 | 100 |
| 129155-8 | 0.24 | 100 | 0.07 | 100 | 0.11 | 100 |
| CDC337 | 0.24 | 100 | 0.07 | 100 | 0.11 | 100 |
| NYH 209 | 0.24 | 100 | 0.07 | 100 | 0.11 | 100 |
| PRA-98 | 0.24 | 100 | 0.07 | 100 | 0.11 | 100 |
| 801805 | 0.24 | 100 | 0.07 | 100 | 0.11 | 100 |
| BACT-053LR01 | 0.24 | 100 | 0.07 | 100 | 0.11 | 100 |

* Contrived vaginal swab matrix was used to represent vaginal and endocervical swab specimens.

| Strain | Swabs* | | Urine Specimens | | Meatal Swab | | | |
|--------|-----------|-------|-----------------|-------|-------------|-------|--|--|
| | copies/mL | % Pos | copies /mL | % Pos | copies /mL | % Pos | | |
| SEA-1 | 5.0 | 100 | 0.8 | 95.8 | 0.5 | 100 | | |
| M2288 | 5.0 | 100 | 0.8 | 100 | 0.5 | 100 | | |
| M2300 | 5.0 | 100 | 0.8 | 100 | 0.5 | 83.3 | | |
| M2321 | 5.0 | 100 | 1.6 | 100 | 1.0 | 87.5 | | |
| M2341 | 5.0 | 100 | 0.8 | 95.8 | 0.5 | 95.8 | | |

Table 4: LoD verification MG strains

* Contrived vaginal swab matrix was used to represent vaginal and endocervical swab specimens.

### 4.3. Precision (within laboratory)

In-house precision was examined using a panel composed of TV and MG cultures diluted into pooled negative urine stabilized in cobas® PCR Media, as well as in contrived matrices representing vaginal and meatal swab specimens collected in cobas® PCR Media or in cervical specimens collected in PreservCyt® Solution.

The precision panel for each matrix was designed to include members without TV and/or MG as well as those with high negative, low and moderate concentrations of TV and MG, corresponding to ~0.3x, ~1x and ~3x LoD. Testing was performed using three lots of cobas® TV/MG reagents and two instruments over twelve days and with two runs per day for a total of twenty-four runs. Each run consisted of three replicates of each sample. Description of the precision panels and the observed positivity rates are shown in Table 5. All negative panel members tested negative throughout the study.

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| Level | N Tested | N positive TV | N positive MG | Hit Rate TV | Hit Rate MG | 95% Confidence Interval TV | | 95% Confidence Interval MG | |
|-----------------------------------------------------|----------|---------------|---------------|-------------|-------------|----------------------------|-------------|----------------------------|-------------|
| | | | | | | Lower Limit | Upper Limit | Lower Limit | Upper Limit |
| Swabs collected in cobas® PCR Media | | | | | | | | | |
| Negative | 72 | 0 | 0 | 0.0% | 0.0% | 0.0% | 5.0% | 0.0% | 5.0% |
| High Negative | 72 | 48 | 61 | 66.7% | 84.7% | 54.6% | 77.3% | 74.3% | 92.1% |
| Low | 71 | 69 | 70 | 97.2% | 98.6% | 90.2% | 99.7% | 92.4% | 100% |
| Moderate | 72 | 72 | 72 | 100% | 100% | 95.0% | 100% | 95.0% | 100% |
| Urine stabilized in cobas® PCR Media | | | | | | | | | |
| Negative | 72 | 0 | 0 | 0.0% | 0.0% | 0.0% | 5.0% | 0.0% | 5.0% |
| High Negative | 72 | 44 | 53 | 61.1% | 73.6% | 48.9% | 72.4% | 61.9% | 83.3% |
| Low | 72 | 72 | 72 | 100% | 100% | 95.0% | 100% | 95.0% | 100% |
| Moderate | 72 | 72 | 72 | 100% | 100% | 95.0% | 100% | 95.0% | 100% |
| Meatal Swab collected in cobas® PCR Media | | | | | | | | | |
| Negative | 72 | N/A | 0 | N/A | 0.0% | N/A | N/A | 0.0% | 5.0% |
| High Negative | 72 | N/A | 41 | N/A | 56.9% | N/A | N/A | 44.7% | 68.6% |
| Low | 72 | N/A | 69 | N/A | 95.8% | N/A | N/A | 88.3% | 99.1% |
| Moderate | 72 | N/A | 72 | N/A | 100% | N/A | N/A | 95.0% | 100% |
| Cervical specimens collected in PreservCyt Solution | | | | | | | | | |
| Negative | 72 | 0 | N/A | 0.0% | N/A | 0.0% | 5.0% | N/A | N/A |
| High Negative | 72 | 39 | N/A | 54.2% | N/A | 42.0% | 66.0% | N/A | N/A |
| Low | 72 | 69 | N/A | 95.8% | N/A | 88.3% | 99.1% | N/A | N/A |
| Moderate | 72 | 72 | N/A | 100% | N/A | 95.0% | 100% | N/A | N/A |

### Table 5: Summary of within laboratory precision

*N/A = Not Applicable

Analysis of standard deviation and percent coefficient of variation of the Ct values from valid tests performed on positive panel members (see Table 6 and Table 7) yielded overall CV (%) ranges from 1.5% to 2.8% for TV and from 1.2% to 4.9% for MG.

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### Table 6: Overall mean, standard deviations and coefficients of variation (%) for cycle threshold, TV positive panels

| Level | Hit Rate | Mean<br>Ct | Within run | Between run | | Between day | | Between<br>instrument | | Between lot | | Total | | |
|------------------------------------------------------|----------|------------|------------|-------------|------|-------------|------|-----------------------|------|-------------|------|-------|------|-----|
| | | | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% |
| Swabs collected in cobas® PCR Media | | | | | | | | | | | | | | |
| High Negative | 66.7% | 37.6 | 0.98 | 2.6 | 0.0 | 0.0 | 0.0 | 0.0 | 0.26 | 0.7 | 0.22 | 0.7 | 1.04 | 2.8 |
| Low | 97.2% | 36.5 | 0.62 | 1.7 | 0.22 | 0.6 | 0.00 | 0.0 | 0.60 | 1.6 | 0.19 | 0.5 | 0.91 | 2.5 |
| Moderate | 100.0% | 35.5 | 0.38 | 1.1 | 0.05 | 0.2 | 0.03 | 0.1 | 0.74 | 2.1 | 0.15 | 0.4 | 0.85 | 2.4 |
| Urine stabilized in cobas® PCR Media | | | | | | | | | | | | | | |
| High Negative | 61.1% | 37.7 | 0.86 | 2.3 | 0.00 | 0.0 | 0.25 | 0.7 | 0.00 | 0.0 | 0.10 | 0.3 | 0.90 | 2.4 |
| Low | 100.0% | 36.7 | 0.62 | 1.7 | 0.31 | 0.8 | 0.18 | 0.5 | 0.11 | 0.3 | 0.16 | 0.4 | 0.74 | 2.0 |
| Moderate | 100.0% | 35.6 | 0.36 | 1.0 | 0.09 | 0.3 | 0.14 | 0.4 | 0.33 | 0.9 | 0.11 | 0.3 | 0.53 | 1.5 |
| Cervical specimens collected in PreservCyt® Solution | | | | | | | | | | | | | | |
| High Negative | 54.2% | 37.6 | 0.65 | 1.7 | 0.30 | 0.8 | 0.29 | 0.8 | 0.42 | 1.1 | 0.00 | 0.0 | 0.87 | 2.3 |
| Low | 95.8% | 36.7 | 0.69 | 1.9 | 0.28 | 0.8 | 0.00 | 0.0 | 0.50 | 1.4 | 0.06 | 0.2 | 0.90 | 2.4 |
| Moderate | 100.0% | 35.6 | 0.64 | 1.8 | 0.15 | 0.4 | 0.00 | 0.0 | 0.64 | 1.8 | 0.00 | 0.0 | 0.92 | 2.6 |

### Table 7: Overall mean, standard deviations and coefficients of variation (%) for cycle threshold, MG positive panels

| Level | Hit Rate | Mean<br>Ct | Within run | | Between run | | Between<br>day | | Between<br>instrument | | Between lot | | Total | |
|--------------------------------------|----------|------------|------------|-----|-------------|-----|----------------|---------|-----------------------|-----|-------------|-----|-------|-----|
| | | | SD | CV% | SD | CV% | SD | CV<br>% | SD | CV% | SD | CV% | SD | CV% |
| Swabs collected in cobas® PCR Media | | | | | | | | | | | | | | |
| High Negative | 84.7% | 37.2 | 1.29 | 3.5 | 0.00 | 0.0 | 0.00 | 0.0 | 0.98 | 2.6 | 0.00 | 0.0 | 1.62 | 4.3 |
| Low | 98.6% | 35.6 | 0.56 | 1.6 | 0.00 | 0.0 | 0.16 | 0.5 | 0.71 | 2.0 | 0.05 | 0.1 | 0.92 | 2.6 |
| Moderate | 100.0% | 34.7 | 0.26 | 0.7 | 0.00 | 0.0 | 0.05 | 0.1 | 0.73 | 2.1 | 0.10 | 0.3 | 0.78 | 2.3 |
| Urine stabilized in cobas® PCR Media | | | | | | | | | | | | | | |
| High Negative | 73.6% | 37.9 | 1.19 | 3.2 | 0.00 | 0.0 | 0.00 | 0.0 | 0.00 | 0.0 | 0.32 | 0.8 | 1.24 | 3.3 |
| Low | 100.0% | 36.3 | 0.66 | 1.8 | 0.21 | 0.6 | 0.00 | 0.0 | 0.25 | 0.7 | 0.20 | 0.6 | 0.76 | 2.1 |
| Moderate | 100.0% | 35.2 | 0.25 | 0.7 | 0.18 | 0.5 | 0.00 | 0.0 | 0.28 | 0.8 | 0.09 | 0.3 | 0.42 | 1.2 |

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| Meatal Swab collected in <b>cobas</b> ® PCR Media | | | | | | | | | | | | | | |
|---------------------------------------------------|--------|------|------|-----|------|-----|------|-----|------|-----|------|-----|------|-----|
| High Negative | 56.9% | 38.1 | 1.55 | 4.1 | 0.37 | 1.0 | 0.00 | 0.0 | 0.95 | 2.5 | 0.00 | 0.0 | 1.85 | 4.9 |
| Low | 95.8% | 37.0 | 0.78 | 2.1 | 0.00 | 0.0 | 0.00 | 0.0 | 0.39 | 1.1 | 0.00 | 0.0 | 0.87 | 2.4 |
| Moderate | 100.0% | 35.7 | 0.33 | 0.9 | 0.00 | 0.0 | 0.00 | 0.0 | 0.32 | 0.9 | 0.18 | 0.5 | 0.50 | 1.4 |

#### Analytical specificity/cross reactivity 4.4.

A panel of 102 bacteria, fungi and viruses, including those commonly found in the male and female urogenital tract, were tested with cobas® TV/MG to assess analytical specificity. The organisms listed in Table 8 were spiked at concentrations of approximately 1 x 10° units/mL for bacteria and approximately 1 x 105 units/mL for viruses into pooled negative urine stabilized in cobas® PCR Media. Testing was performed with each potential interfering organism in absence and presence of TV and MG target (spiked at approximately 3x LoD). Three replicates were tested for each organism in absence of target and one in presence of target. None of the organisms tested interfered with the test performance by generating false positive results for either TV or MG targets. Detection of MG target was not affected by any of the organisms tested. Trichomonas tenax interfered with detection of TV target at concentration level of 1 x 10° CFU/mL, but did not interfere with detection of TV target when tested at concentration level of 1 x 104 CFU/mL. Trichomonas tenax is a commensal of the oral cavity.

| Microorganism | Microorganism | Microorganism |
|------------------------------|------------------------------|-------------------------------|
| Acholeplasma laidlawii | Enterococcus avium | Mycoplasma faucium |
| Acholeplasma oculi | Enterococcus faecalis | Mycoplasma fermentans |
| Achromobacter xerosis | Enterococcus faecium | Mycoplasma hominis |
| Acinetobacter Iwoffii | Erysipelothrix rhusiopathiae | Mycoplasma orale |
| Actinomyces israelii | Escherichia coli | Mycoplasma penetrans |
| Aerococcus viridans | Flavobacterium | Mycoplasma pirum |
| Aeromonas hydrophila | Fusobacterium nucleatum | Mycoplasma pneumoniae |
| Alcaligenes faecalis | Gardnerella vaginalis | Mycoplasma primatum |
| Atopobium vaginae | Gemella haemolysans | Mycoplasma salivarium |
| Bacillus subtilis | Giardia intestinalis | Mycoplasma spermatophilum** |
| Bacteroides fragilis | Haemophilus ducreyi | Neisseria gonorrhoeae |
| Bacteroides ureolyticus | Herpes Simplex Virus Type 1* | Pentatrichomonas hominis |
| Bifidobacterium adolescentis | Herpes Simplex Virus Type 2* | Peptostreptococcus anaerobius |

Microorganisms tested for analytical specificity/cross reactivity Table 8:

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| Microorganism | Microorganism | Microorganism |
|----------------------------|--------------------------------|----------------------------|
| Branhamella catarrhalis | Mycoplasma hominis | Prevotella bivia |
| Brevibacterium linens | Human Immunodeficiency Virus* | Propionibacterium acnes |
| Campylobacter jejuni | Human Papillomavirus type 16 | Proteus mirabilis |
| Candida albicans | Kingella denitrificans | Providencia stuartii |
| Candida glabrata | Klebsiella oxytoca | Pseudomonas aeruginosa |
| Candida parapilosis | Klebsiella pneumoniae | Rahnella aquatilis |
| Candida tropicalis | Lactobacillus acidophilus | Rhizobium radiobacter |
| Chlamydia trachomatis | Lactobacillus crispatus | Rhodospirillum rubrum |
| Chromobacterium violaceum | Lactobacillus jensenii | Saccharomyces cerevisiae |
| Citrobacter braakii | Lactobacillus vaginalis | Salmonella minnesota |
| Clostrodium perfringens | Leptotrichia buccalis | Serratia marcescens |
| Clostridioides difficile** | Leuconostoc mesenteroides | Staphylococcus aureus |
| Corynebacterium genitalium | Leuconostoc paramensenteroides | Staphylococcus epidermidis |
| Corynebacterium xerosis | Listeria monocytogenes | Streptococcus agalactiae |
| Cryptococcus neoformans | Micrococcus luteus | Streptococcus pneumoniae |
| Cytomegalovirus | Mobiluncus curtisii | Streptococcus pyogenes |
| Derxia gummosa | Moraxella osloensis | Trichomonas tenax*** |
| Dientamoeba fragilis | Morexella catarrhalis | Ureaplasma urealyticum**** |
| Eikenella corrodens | Morexella lacunata | Veillonella parvula |
| Enterobacter aerogenes | Morganella morganii | Vibrio parahaemolyticus |
| Enterobacter cloacae | Mycobacterium smegmatis | Yersinia enterocolitica |

Unless noted (below), bacteria and fungi were quantified as Colony Forming Units (CFU) and viruses were quantified as International Units (IU).

* Quantified in copies/mL

** Previously known as Clostridium difficile

***Interference with TV detection observed when tested at 1 x106 CFU/mL. No interference with TV detection observed when tested at 1 x 104 CFU/mL or below.

****Quantified in color changing units (ccu)

#### 4.5. Interference

The effect of over-the-counter or prescription products that may be present in urogenital specimens (Table 9) were evaluated. Possible interference from glacial acetic acid occasionally utilized in cytologic evaluation of cervical specimens was also assessed. Testing was done using pooled negative clinical specimens in the presence of TV and MG (spiked at approximately 3x LoD), and non-clinical/contrived matrices when testing in the absence of TV and MG. For each specimen type, three replicates were tested for each substance in the presence of and one in the absence of target organisms.

Eighteen products (including glacial acetic acid) as listed in Table 9 did not interfere with cobas® TV/MG when tested at concentrations of 1.0mg/mL or 1% v/v as applicable.

{16}------------------------------------------------

### List of products that do not interfere with test performance in urogenital Table 9: specimens

| Product Name | | |
|---------------------------------------------------|-------------------------------------------|------------------------------------|
| Clindamycin Phosphate Vaginal Cream | Monistat® Complete Care Itch Relief Cream | Yeast Guard Advanced |
| CVS tioconazole 1 (Equate tioconazole 1) | Gyne-Lotrimin 7 | Glacial acetic acid |
| Equate Vagicaine Anti-Itch Cream | Norforms Suppositories | Azo Standard |
| Estrace | Premarin | Arilin rapid vaginal suppositories |
| K-Y™ UltraGel (Replaces KY Silk E) | Summer's Eve Feminine Deodorant Spray | Vagi Metro Cream |
| Monistat 3 Vaginal Antifungal Combination<br>Pack | Vaginal Contraceptive Foam | Nidazea Gel |

Only Replens™, RepHresh™ Clean Balance, and Metronidazole Vaginal Gel by Sandoz interfered with cobas® TV/MG by producing false negative or invalid results at the concentrations above those stated in Table 10. It should be noted that three other products containing metronidazole (Arilin rapid vaginal suppositories, Vagi Metro Cream and Nidazea Gel), did not cause interference (Table 9).

Table 10: List of products that interfere with test performance above the concentration stated

| Product Name | Swabs* | Urine Specimens | Meatal Swab | PreservCyt®<br>Specimens |
|----------------------------------------------|--------|-----------------|-------------|--------------------------|
| | mg/mL | mg/mL | mg/mL | mg/mL |
| Replens™ Long-Lasting Vaginal<br>Moisturizer | 1.0 | 0.5 | 0.3 | 2.0 |
| RepHresh™ Clean Balance | 2.0 | 1.0 | 0.5 | 2.0 |
| Metronidazole Vaginal Gel by<br>Sandoz | 1.0 | 0.2 | 0.3 | 1.0 |

* Vaginal swab samples were used as a representative swab sample type for vaginal and endocervical swab specimens.

Endogenous substances that may be present in urogenital specimens were tested for interference. Testing was done using pooled negative clinical specimens in the presence of TV and MG (spiked at approximately 3x LoD), and non-clinical/contrived matrices when testing in the absence of TV and MG. At least twelve replicates were tested in total for each substance per condition across specimen types where the endogenous substances are expected to be present.

{17}------------------------------------------------

None of the substances interfered with the test performance by generating falsepositive results. Levels of endogenous substances tolerated by the assay for all specimen types are shown in Table 11.

| Interferent | Swabs** | Meatal Swab | PreservCyt®<br>Specimens | Urine |
|---------------------------------------|------------------|-------------|--------------------------|------------------|
| Albumin (% w/v) | N/A | N/A | N/A | 0.5% |
| Bilirubin (% w/v) | N/A | N/A | N/A | 1.0% |
| Mucus* | present | present | present | present |
| Glucose (% w/v) | N/A | N/A | N/A | 1.0 % |
| Peripheral Blood Mononuclear<br>Cells | 1.0E+06 cells/mL | N/A | 1.0E+06 cells/mL | 1.0E+06 cells/mL |
| pH (acidic and alkaline) | N/A | N/A | N/A | pH 4 and pH 9 |
| Semen | 22 mg/mL | 20 mg/mL | 4 mg/mL | 13 mg/mL |
| Whole Blood (% v/v) | 10% | N/A | 10% | 10% |

Table 11: Summary of endogenous substance concentrations that do not show interference

* One mucus swab per sample reflecting the maximum level that could be found in patient sample.

**Endocervical swab samples were used as a representative swab sample type for vaginal and endocervical swab specimens.

***Semen tested from swab dipped in fluid. Swab was weighed before and after to determine concentration.

#### Competitive inhibition 4.6.

To assess competitive inhibition between TV and MG, samples of swabs, urine and PreservCvt® specimens were tested with low and moderate concentrations of…