APTIMA TRICHOMONAS VAGINALIS ASSAY - PANTHER

K122062 · Gen-Probe, Inc. · OUY · Jan 9, 2013 · Immunology

Device Facts

• Record ID: K122062
• Device Name: APTIMA TRICHOMONAS VAGINALIS ASSAY - PANTHER
• Applicant: Gen-Probe, Inc.
• Product Code: OUY · Immunology
• Decision Date: Jan 9, 2013
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3860
• Device Class: Class 2

Intended Use

The APTIMA Trichomonas vaginalis Assay is an in vitro qualitative nucleic acid amplification test (NAAT) for the detection of ribosomal RNA (rRNA) from Trichomonas vaginalis to aid in the diagnosis of trichomoniasis using the PANTHER System. The assay may be used to test the following specimens from symptomatic or asymptomatic women: clinician-collected endocervical swabs, clinician-collected vaginal swabs, and specimens collected in PreservCyt Solution.

Device Story

The APTIMA Trichomonas vaginalis Assay is a qualitative NAAT performed on the automated PANTHER System. It processes clinician-collected vaginal swabs, endocervical swabs, or PreservCyt liquid Pap specimens. The assay uses Target Capture (TC) with magnetic microparticles to isolate rRNA, followed by Transcription Mediated Amplification (TMA) to generate RNA amplicons. Detection is achieved via Hybridization Protection Assay (HPA) using acridinium ester-labeled DNA probes; light emission is measured as Relative Light Units (RLU) in a luminometer. The PANTHER System automates specimen handling, amplification, and detection. Results are interpreted by the system software as negative, positive, or invalid based on RLU thresholds. The device aids clinicians in diagnosing trichomoniasis, facilitating appropriate patient management.

Clinical Evidence

Clinical performance was evaluated using 689 vaginal, 737 endocervical, and 791 PreservCyt specimens from symptomatic and asymptomatic women. Infected status was determined by a reference algorithm (culture/wet mount). Sensitivity was 100% across all specimen types. Specificity ranged from 98.1% to 98.6%. Analytical studies included reproducibility (108 replicates per panel member), precision (162 replicates per panel member), and carryover (0% rate). No significant cross-reactivity was observed with 42 tested microorganisms, except for rare T. tenax and P. hominis.

Technological Characteristics

Nucleic acid amplification test (NAAT) using Target Capture, TMA, and HPA. Reagents include HEPES-buffered solution, lithium salts, and magnetic beads. Automated platform (PANTHER) performs specimen processing, amplification, and detection. Detection via chemiluminescent acridinium ester-labeled DNA probes measured as RLU. Software-based automated interpretation of results. Complies with EN 13640, CLSI EP5-A2, EP15-A2, and EP07-A2 standards.

Indications for Use

Indicated for symptomatic or asymptomatic women to aid in the diagnosis of trichomoniasis via detection of T. vaginalis rRNA in clinician-collected endocervical swabs, vaginal swabs, or PreservCyt Solution specimens.

Regulatory Classification

Identification

A Trichomonas 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 by Trichomonas vaginalis.

Special Controls

*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 of*Trichomonas vaginalis;* Guideline for Industry and Food and Drug Administration Staff.” See § 866.1(e) for information on obtaining this document.

Predicate Devices

• APTIMA Trichomonas vaginalis Assay on the TIGRIS DTS System (K102911)

Submission Summary (Full Text)

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## SECTION 2 510(k) SUMMARY

# JAN 0 9 2013

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of 21 CFR 807.92. The information provided is consistent with FDA recommendations cited in Appendix B – The 510(k) Summary Document Requirements from the draft guidance document entitled, "The 510(k) Program: Evaluating Substantial Equivalence in Premarket Notifications [510(k)]" (Dec. 27, 2011).

## A. 510(k) Number:

K122062

### B. Purpose of Submission:

Device Modification to K102911: Change in formulation of the Target Capture Reagent (TCR) used in APTIMA Trichomonas vaginalis (ATV) Assay, and use of the PANTHER System as a platform.

## C. Measurand:

Ribosomal RNA from Trichomonas vaginalis

## D. Type of Test:

Nucleic acid amplification test

### E. Applicant:

Gen-Probe Incorporated 10210 Genetic Center Drive San Diego, CA 92121 [phone]

| Company Contact: | Ma. Carmelita S. Baluyot<br>Regulatory Affairs Specialist |
|------------------|-----------------------------------------------------------|
| Phone: | 858-410-8309 |
| Fax: | 858-410-9028 |
| Email: | [e-mail] |
| Date Prepared: | December 18, 2012 |

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## F. Proprietary and Established Names:

APTIMA Trichomonas vaginalis Assay

## G. Regulatory Information

- 1. Regulation Section
- 21 CFR 866.3860
- 2. Classification Class II
- 3. Product Code OUY - Trichomonas vaginalis nucleic acid amplification test system
- 4. Panel 83 - Microbiology

## H. Intended Use

- l. Intended Use
The APTIMA Trichomonas vaginalis Assay is an in vitro qualitative nucleic acid amplification test (NAAT) for the detection of ribosomal RNA (rRNA) from Trichomonas vaginalis to aid in the diagnosis of trichomoniasis using the PANTHER System.

The assay may be used to test the following specimens from symptomatic or asymptomatic women: clinician-collected endocervical swabs, clinician-collected vaginal swabs, and specimens collected in PreservCyt Solution.

### 2. Indications for Use

The APTIMA Trichomonas vaginalis Assay is an in vitro qualitative nucleic acid amplification test (NAAT) for the detection of ribosomal RNA (rRNA) from Trichomonas vaginalis to aid in the diagnosis of trichomoniasis using the PANTHER System.

The assay may be used to test the following specimens from symptomatic or asymptomatic women: clinician-collected endocervical swabs, clinician-collected vaginal swabs, and specimens collected in PreservCyt Solution.

- 3. Special conditions for use statement
For prescription use

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## 4. Special instrument requirements

The automated PANTHER System

### I. Device Description

The ATV Assay with the modified TCR, referred to as ATV Assay (Version 2) in this submission is the subject of this premarket notification. The ATV Assay (Version 2) is similar to the ATV Assay originally cleared (ref: K102911), except for the formulation of the TCR. The TCR is a HEPES-buffered solution containing lithium salts and derivatized magnetic beads. A second target capture oligo was added to the TCR formulation in order to accommodate future specimen types.

The TCR modification did not result in the change of assay technology. The ATV Assay (Version 2) uses Target Capture (TC), Transcription Mediated Amplification (TMA), and Hybridization Protection Assay (HPA) technologies to qualitatively detect ribosomal RNA (rRNA) from Trichomonas vaginalis. The overall assay design as well as the assay procedural steps remain unchanged from that previously described in the original 510(k) for the ATV Assay (K102911).

The ATV Assay (Version 2) kit is comprised of 3 boxes:

- 1. Refrigerated Box Contains the Amplification Reagent, Enzyme Reagent, Probe Reagent and Target Capture Reagent-B
- 2. Room Temperature Box Contains Amplification Reconstitution Solution, Enzyme Reconstitution Solution, Probe Reconstitution Solution, Selection Reagent and Target Capture Reagent
- 3. Controls Box Contains the Negative and Positive Controls

The ATV Assay (Version 2) on PANTHER would utilize three specimen collection kits. These collection kits were cleared for use with the originally cleared ATV Assay and other commercialized APTIMA Assays.

- 1. APTIMA Unisex Swab Specimen Collection Kit for Endocervical and Male Urethral Swab Specimens
- 2. APTIMA Vaginal Swab Specimen Collection Kit
- 3. APTIMA Specimen Transfer Kit

### Instrumentation

The ATV Assay (Version 2) was validated using the PANTHER System, which was previously cleared in May 2012 (Ref: K111409).

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## J. Substantial Equivalence Information:

- A. Predicate Device Name:
- APTIMA Trichomonas vaginalis Assay on TIGRIS
- B. Predicate 510(k) Number:

K102911

- C. Comparison with Predicate:

| Similarities | | |
|------------------------------------|---------------------------------------------------------------------------------------------------------------|----------------------------------|
| Item | ATV Assay (Version 2)<br>Device | ATV Assay (K102911)<br>Predicate |
| Device Class | 2 | same |
| Qualitative<br>/Quantitative Assay | Qualitative | same |
| Intended Use | NAAT test for detection of<br>Trichomonas vaginalis<br>ribosomal RNA (rRNA) | same |
| Technology | Target Capture(TC)<br>Transcription Mediated<br>Amplification(TMA)<br>Hybridization Protection<br>Assay (HPA) | same |

| Differences | | |
|----------------|----------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------|
| Item | ATV Assay (Version 2) Device | ATV Assay (K102911) Predicate |
| Formulation | Original Formulation plus additional oligo in TCR | Original Formulation |
| Platform | PANTHER | TIGRIS |
| Specimen Types | Three (3) Female specimen types:<br>• Vaginal swab<br>• Endocervical swab<br>• ThinPrep in PreservCyt solution | Four (4) Female specimen types:<br>• Urine<br>• Vaginal swab<br>• Endocervical swab<br>• ThinPrep in PreservCyt solution |

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## K. Standard/ Guidance Document Referenced

EN 13640:2002 - Stability Testing of In-Vitro Diagnostic Medical Devices

EP5-A2, 2004 - Evaluation of Precision Performance of Quantitative Measurement Methods, CLSI Approved Guideline

EP15-A2, 2006 - User Verification of Performance for Precision and Trueness, CLSI Approved Guideline

EP07-A2, 2005 - Interference Testing in Clinical Chemistry, Approved Guideline

Format for Traditional and Abbreviated 510(k)s - Guidance for Industry and FDA Staff, August 2005

General Principles of Software Validation, Final Guidance for Industry and FDA Staff, January 2002

The 510(k) Program: Evaluating Substantial Equivalence in Premarket Notifications [510(k)], Draft Guidance for Industry and FDA Staff, Dec. 27, 2011.

### L. Test Principle

The ATV Assay involves 3 main steps which take place in a single tube: target capture (TC), target amplification by Transcription Mediated Amplification (TMA) and detection of the amplification products (amplicon) by Hybridization Protection Assay (HPA). Specimens to be tested are collected and transferred into their respective specimen transport tubes. The transport solutions in the specimen transport tubes release the rRNA targets and protect them from degradation. When the TV assay is performed, the target rRNA is isolated from the specimen by use of capture oligomers via target capture that utilizes magnetic particles. When target capture is complete, the TV rRNA is amplified via TMA. Detection of the amplicon is achieved by HPA using single stranded nucleic acid probes with chemiluminescent labels that are complimentary to the amplicon. During the detection step, light emitted from the labeled RNA: DNA hybrids is measured as photon signals in a luminometer and are reported as Relative Light Units (RLU).

## M. Performance Characteristics

1. Analytical Performance

#### Precision/Reproducibility a.

The precision of the ATV Assay (Version.2) on PANTHER was evaluated in-house by multiple operators who performed multiple runs over multiple days with three different reagent lots on three different PANTHER Systems.

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A 4-member reproducibility panel was prepared in PreservCyt (ThinPrep) liquid cytology specimen-STM matrix (1:2.9). PreservCyt sample pools were screened with the ATV Assay to confirm that 7. vaginalis was not present. The pools were then spiked with T. vaginalis lysate to produce high negative (expected positivity: >0% and <100%), moderate positive (expected positivity: 100%) and high positive (expected positivity: 100%) panel members. Unspiked PreservCyt pools served as the negative panel members. The reproducibility panels were tested in triplicate by 3 operators, in 2 runs per day using 3 reagent lots on 3 instruments. Testing was performed over 13 days.

The negative and positive panel members (panel members 1, 3, and 4) vielded ≥99% agreement with the expected results as shown by the following chart. The results from this study demonstrate that the ATV Assay can be performed reproducibly on the PANTHER System.

| Description | Panel<br>Member | TV/mL | Valid N | %<br>Positive | % Agreement |
|-------------------|-----------------|-------|---------|---------------|-------------------|
| Negative | 1 | 0 | 162 | 0.0 | 100% (97.7 - 100) |
| High Negative | 2 | 0.003 | 162 | 11.1 | N/A |
| Moderate Positive | 3 | 0.02 | 162 | 99.4 | 99% (96.6 -99.9) |
| High Positive | 4 | 1 | 162 | 100.0 | 100% (97.7 - 100) |

N/A = not applicable, expected positivity was 5% to 95%. N= 162 for each panel member

#### b. Assay reportable range

The ATV Assay (Version 2) is designed for and validated on the PANTHER System. The assay test results are automatically interpreted by the PANTHER System APTIMA Trichomonas vaginalis software. A test result may be negative, positive or invalid as determined by the total Relative Light Units (RLU) in the detection step. A test result may be invalid due to RLU values outside the normal expected ranges. Initial invalid results should be retested.

| Test Interpretation | Total RLU (x 1000) |
|---------------------|--------------------|
| Negative | 0* to <100 |
| Positive | 100 to <2400 |
| Invalid | 0* or >/= 2400 |

*If the RLU measured on the PANTHER System is between 0 and 999, a result of "O" is reported in the "Total RLU (000s)" column in the run report. Measured RLU values less than 690 are reported as invalid. RLU values between 690 and 999 are reported as valid.

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ﻥ ﺍ Traceability, Stability, Expected Values (controls, calibrators or methods)

Data to support the recommended shipping and storage conditions for the vaginal swab and PreservCyt liquid Pap were generated with negative clinical specimens spiked with T. vaginalis. Greater than 98% positivity was observed in both matrices at all times and temperatures tested confirming the validity of the claimed maximum storage times and temperatures.

## Quality Control Results and Acceptability

The APTIMA Negative Control for Trichomonas and APTIMA Positive Control for Trichomonas act as controls for the target capture, amplification and detection steps of the assay. The Positive Control contains non-infectious Trichomonas vaginalis rRNA.

| Control | Total RLU (x1000) | Trichomonas vaginalis<br>Result |
|------------------|-------------------|---------------------------------|
| Negative Control | 0* and <20 | Negative |
| Positive Control | >/=500 and < 2400 | Positive |

The ATV Assay Controls must produce the following test results:

*If the RLU measured on the PANTHER System in between 0 and 999, a result of "O" is reported in the "Total RLU (000s)" column in the run report. Measured RLU values less than 690 are reported as invalid. RLU values between 690 and 999 are reported as valid.

#### d. Analytical Specificity

Specificity of the ATV Assay (Version 2) was evaluated by testing various microorganisms, including common flora of the genitourinary tract, opportunistic organisms, and closely related organisms. Testing was conducted in specimen transport media (STM) and PreservCyt in STM with 25 replicates of each isolate. The list of organisms and the concentrations tested are provided in Table 1. No cross-reactivity or significant effect on ATV Assay specificity was observed with any of the organisms tested.

The ATV Assay (Version 2) was also evaluated by testing the same organisms (Table 1) in STM and PreservCyt in STM spiked with T. vaginalis lysate to a final concentration of 0.01 TV/mL (25 replicates of each isolate). The ATV Assay was not significantly affected by the presence of the microorganisms tested, except in the presence of Trichomonas tenax and Pentatrichomonas hominis (where lower signal outputs were observed). T. tenax is a commensal of the oral cavity and P. hominis is a commensal of the large intestine.

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| | | | | | Table 1 - Microorganisms tested with ATV Assay on PANTHER | | | | | | | | | |
|--|--|--|--|--|-----------------------------------------------------------|--|--|--|--|--|--|--|--|--|
|--|--|--|--|--|-----------------------------------------------------------|--|--|--|--|--|--|--|--|--|

| Microorganism | Concentration | Microorganism | Concentration |
|------------------------------|-------------------|----------------------------|--------------------|
| Acinetobacter Iwoffi | 1x106 CFU/mL | HPV 16 | 2.5x106 copies/mL |
| Actinomyces israelii | 1x106 CFU/mL | HPV 6 | 2.5x106 copies/mL |
| Atopobium vaginae | 1x106 CFU/mL | Klebsiella pneumoniae | 1x106 CFU/mL |
| Bacteroides fragilis | 1x106 CFU/mL | Lactobacillus acidophilus | 1x106 CFU/mL |
| Bifidobacterium adolescentis | 1x106 CFU/mL | Lactobacillus crispatus | 1x106 CFU/mL |
| Campylobacter jejuni | 1x106 CFU/mL | Listeria monocytogenes | 1x106 CFU/mL |
| Candida albicans | 1x106 CFU/mL | Mobiluncus curtisii | 1x106 CFU/mL |
| Chlamydia trachomatis | 1x106 IFU/mL | Mycoplasma genitalium | 2.5 x106 copies/mL |
| Clostridium difficile | 1x106 CFU/mL | Mycoplasma hominis | 1x106 CFU/mL |
| Corynebacterium genitalium | 1x106 CFU/mL | Neisseria gonorrhoeae | 1x106 CFU/mL |
| Cryptococcus neoformans | 1x106 CFU/mL | Pentatrichomonas hominis | 1x106 cells/mL |
| Cytomegalovirus | 2x105 TCID50/mL | Peptostreptococcus magnus | 1x106 CFU/mL |
| Dientamoeba fragilis | 1x106 CFU/mL | Prevotella bivia | 1x106 CFU/mL |
| Enterobacter cloacae | 1x106 CFU/mL | Propionibacterium acnes | 1x106 CFU/mL |
| Enterococcus faecalis | 1x106 CFU/mL | Proteus vulgaris | 1x106 CFU/mL |
| Escherichia coli | 1x106 CFU/mL | Pseudomonas aeruginosa | 1x106 CFU/mL |
| Gardnerella vaginalis | 1x106 CFU/mL | Staphylococcus aureus | 1x106 CFU/mL |
| Haemophilus ducreyi | 1x106 CFU/mL | Staphylococcus epidermidis | 1x106 CFU/mL |
| Herpes simplex virus I | 2x105 TCID50/mL | Streptococcus agalactiae | 1x106 CFU/mL |
| Herpes simplex virus II | 2x105 TCID50/mL | Trichomonas tenax | 1x106 cells/mL |
| HIV-1 | 2.5x106 copies/mL | Ureaplasma urealyticum | 1x106 CFU/mL |

#### Interference e.

The following substances were individually spiked into STM and PreservCyt in STM for a final concentration of 1% (vol/vol or wt/vol): personal lubricants, personal deodorants, spermicides, anti-fungals, intravaginal hormones, porcine gastric mucus, seminal fluid from 25 donors, and whole blood (10% final concentration). Glacial acetic acid was tested by spiking into PreservCyt-STM (10% final concentration). Samples with each interfering substance alone as well as samples spiked with 7. vaginalis lysate to a final concentration of 0.01 TV/mL were tested.

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Testing results yielded no false positive results for all substances tested (100% specificity).

No interference with the detection of T. vaginalis at concentrations of 0.01 TV/mL (≥ 95% sensitivity) was observed with any of the substances tested with the exception of Astroglide personal lubricant, porcine gastric mucus, and glacial acetic acid. Astroglide personal lubricant and glacial acetic acid did not interfere with the detection of T. vaginalis when tested at a concentration of 0.3 TV/mL (100% sensitivity). Porcine gastric mucus did not interfere with the detection of T. vaginalis when tested at a concentration of 1 TV/mL (100% sensitivity).

#### f. Detection Limit

Sensitivity panels were prepared with two strains of T. vaginalis (one Metronidazolesusceptible strain and one Metronidazole-resistant strain). Testing showed greater than 95% positivity in both strains of T. vaginalis for panels containing 0.01 TV/mL in PreservCyt liquid Pap specimen matrix and panels containing 0.003 TV/mL in swab specimen matrix.

#### Assay Cut-Off g.

The ATV Assay (Version 2) is designed for and validated on the PANTHER System. The assay test results are automatically interpreted by the PANTHER System APTIMA Trichomonas vaginalis software. A test result may be negative, positive or invalid as determined by the total Relative Light Units (RLU) in the detection step. A test result may be invalid due to RLU values outside the normal expected ranges. Initial invalid results should be retested.

| Test Interpretation | Total RLU (x 1000) |
|---------------------|--------------------|
| Negative | 0* to <100 |
| Positive | 100 to <2400 |
| Invalid | 0* or >/= 2400 |

*If the RLU measured on the PANTHER System is between 0 and 999, a result of "O" is reported in the "Total RLU (000s)" column in the run report. Measured RLU values less than 690 are reported as invalid. RLU values between 690 and 999 are reported as valid.

### 2. Comparison Studies

- Performance characterization relative to an established patient-infected status a. algorithm.
See 3 (a) below

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- b. Matrix comparison
N/A

## 3. Clinical Studies

## a. Clinical Performance (Clinical Sensitivity and Specificity)

Clinical performance of the ATV Assay on the PANTHER System was evaluated using leftover specimens collected from consenting subjects during a previous, prospective, multicenter clinical study of the ATV Assay on the TIGRIS DTS System. Symptomatic and asymptomatic women were enrolled from 9 US clinical sites, including obstetrics and gynecology, family planning, and STD clinics. Three (3) vaginal swabs, 1 endocervical swab, and 1 PreservCyt Solution liquid Pap specimen were collected from each subject. All specimens were clinician-collected.

PreservCyt liquid Pap specimens were collected with a broom-type device or a spatula and cytobrush. Two of the vaginal swab specimens were tested with a commercially available culture system and wet mount microscopic examination to establish infected status. The remaining specimens were prepared for APTIMA Trichomonas vaginalis Assay testing in accordance with the appropriate APTIMA specimen collection kit package insert instructions.

PANTHER System testing with the APTIMA Trichomonas vaginalis Assay was conducted at 3 sites (2 external laboratories and Gen-Probe) in accordance with package insert instructions.

Performance characteristics of the APTIMA Trichomonas vaginalis Assay were estimated by comparing results to a patient infected status algorithm. In the algorithm, the designation of a subject as being infected or non-infected with 7. vaginalis was based on results from vaginal swab specimens tested by culture and/or wet mount microscopic examination. At least one of the reference test results was required to be positive to establish an infected patient status. Both reference tests were required to be negative to establish a non-infected patient status.

Twenty-three (23) APTIMA Trichomonas vaginalis Assay runs were initiated on the PANTHER System. Of these 23 runs, 1 (4.3%, 1/23) was aborted due to a fatal hardware error that led to a software failure. Specimens tested in the aborted run were retested. A total of 689 vaginal swab, 737 endocervical swab, and 791 PreservCyt Solution liquid Pap specimens were tested in the 22 valid runs. Of these specimens, 12 vaginal swab (1.7%, 12/689), 24 endocervical swab (3.3%, 24/737), and 29 PreservCyt Solution liquid Pap (3.7%, 29/791) specimens had initial invalid results due to hardware or software errors. Specimens with initial invalid results were retested. Eleven (11) vaginal swab (1.6%, 11/689), 24 endocervical swab (3.3%, 24/737), and 1 PreservCyt Solution liquid Pap (0.1%, 1/791) specimens had final invalid results due to hardware or software errors; these specimens were excluded from the analyses.

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Results below show the sensitivity, specificity, positive value (PPV), and negative predictive value (NPV) of the ATV Assay and the prevalence of T. vaginalis in each specimen type. Performance was comparable across specimen types.

Table 2 below shows the sensitivity, specificity, PPV, and NPV of the APTIMA Trichomonas vaginalis Assay on the PANTHER System and the prevalence of T. vaginalis (based on the infected status) in each specimen type by symptom status and overall. Subjects were classified as symptomatic if symptoms were reported by the subjects were classified as asymptomatic if the subject did not report symptoms. Prevalence was higher in symptomatic women.

| Specimen<br>Type | Symptom<br>Status | n | TP | FP1 | TN | FN | Prev % | Sensitivity %<br>(95% CI)2 | Specificity %<br>(95% CI)2 | PPV %<br>(95% CI)3 | NPV %<br>(95% CI)3 |
|------------------|-------------------|-----|----|-----|-----|----|--------|----------------------------|----------------------------|---------------------|--------------------|
| CVS | Asymptomatic | 274 | 12 | 7a | 255 | 0 | 4.4 | 100<br>(75.8-100) | 97.3<br>(94.6-98.7) | 63.2<br>(45.8-80.9) | 100<br>(98.9-100) |
| | Symptomatic | 393 | 57 | 4b | 332 | 0 | 14.5 | 100<br>(93.7-100) | 98.8<br>(97.0-99.5) | 93.4<br>(84.9-98.1) | 100<br>(98.9-100) |
| | All | 667 | 69 | 11c | 587 | 0 | 10.3 | 100<br>(94.7-100) | 98.2<br>(96.7-99.0) | 88.3<br>(77.9-92.8) | 100<br>(99.4-100) |
| ES | Asymptomatic | 309 | 16 | 5d | 288 | 0 | 5.2 | 100<br>(80.6-100) | 98.3<br>(96.1-99.3) | 76.2<br>(58.1-90.8) | 100<br>(98.9-100) |
| | Symptomatic | 391 | 51 | 7e | 333 | 0 | 13.0 | 100<br>(93.0-100) | 97.9<br>(95.8-99.0) | 87.9<br>(78.1-94.7) | 100<br>(99.0-100) |
| | All | 700 | 67 | 12f | 621 | 0 | 9.6 | 100<br>(94.6-100) | 98.1<br>(96.7-98.9) | 84.8<br>(76.3-91.5) | 100<br>(99.4-100) |
| PCyt | Asymptomatic | 333 | 19 | 2g | 312 | 0 | 5.7 | 100<br>(83.2-100) | 99.4<br>(97.7-99.8) | 90.5<br>(72.6-98.7) | 100<br>(98.9-100) |
| | Symptomatic | 441 | 64 | 8h | 369 | 0 | 14.5 | 100<br>(94.3-100) | 97.9<br>(95.9-98.9) | 88.9<br>(80.4-94.9) | 100<br>(99.1-100) |
| | All | 774 | 83 | 10i | 681 | 0 | 10.7 | 100<br>(95.6-100) | 98.6<br>(97.4-99.2) | 89.2<br>(82.0-94.5) | 100<br>(99.5-100) |

| Table 2 - PANTHER System: Performance Characteristics of the APTIMA Trichomonas | | |
|---------------------------------------------------------------------------------|--|--|
| vaginalis Assay by Symptom Status | | |

Cl = confidence interval, CVS = dinician-collected vagin.al swab, FN = false negative, FP = false positive, PCyt = PreservCyt Solution liquid Pap, Prev = prevalence, TN = true negative, TP = true positive,

17. vaginalis NAAT results from a previous study (# positive results / # samples tested): a: 47. b: 34. c: 7/11. d 1/5, e: 2/7, f. 3/12 g: 0/2, h: 3/8, i: 3/10.

Score confidence interval

IPPV 95% confidence interval computed from the exact for the positive likelinod rato, NPV 95% confidence interval computed from the exact 95% confidence interval from the negative likelihood ratio

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#### APTIMA Trichomonas vaginalis Assay

Table 3 below shows the sensitivity, specificity, PPV, and NPV of the APTIMA Trichomonas vaginalis Assay on the PANTHER System and the prevalence of T. vaginalis (based on the infected status) in each specimen type by collection site. For each specimen type, performance was similar across collection sites. Prevalence varied across collection sites, as expected.

Table 3: Performance Characteristics of the APTIMA Trichomonas vaginalis Assay by Collection Site

| Site | Specimen<br>Type | n | TP | FP | TN | FN | Prev % | Sensitivity<br>(95% CI)¹ | Specificity<br>(95% CI)¹ | PPV %<br>(95% CI)² | NPV %<br>(95% CI)² |
|------|------------------|-----|----|----|-----|----|--------|--------------------------|--------------------------|--------------------|--------------------|
| 1 | CVS | 52 | 8 | 1 | 43 | 0 | 15.4 | 100 (67.6-100) | 97.7 (88.2-99.6) | 88.9 (60.2-99.7) | 100 (93.7-100) |
| 1 | ES | 53 | 9 | 2 | 42 | 0 | 17.0 | 100 (70.1-100) | 95.5 (84.9-98.7) | 81.8 (56.9-97.4) | 100 (93.5-100) |
| 1 | PCyt | 59 | 11 | 0 | 48 | 0 | 18.6 | 100 (74.1-100) | 100 (92.6-100) | 100 (75.6-100) | 100 (93.9-100) |
| 2 | CVS | 52 | 3 | 1 | 48 | 0 | 5.8 | 100 (43.9-100) | 98.0 (89.3-99.6) | 75.0 (28.5-99.2) | 100 (95.8-100) |
| 2 | ES | 56 | 4 | 1 | 51 | 0 | 7.1 | 100 (51.0-100) | 98.1 (89.0-99.7) | 80.0 (40.5-99.4) | 100 (95.6-100) |
| 2 | PCyt | 68 | 5 | 0 | 63 | 0 | 7.4 | 100 (58.6-100) | 100 (94.3-100) | 100 (58.3-100) | 100 (96.0-100) |
| 3 | CVS | 12 | 2 | 0 | 10 | 0 | 16.7 | 100 (34.2-100) | 100 (72.2-100) | 100 (32.1-100) | 100 (85.6-100) |
| 3 | ES | 16 | 2 | 0 | 14 | 0 | 12.5 | 100 (34.2-100) | 100 (78.5-100) | 100 (31.5-100) | 100 (89.3-100) |
| 3 | PCyt | 17 | 2 | 1 | 14 | 0 | 11.8 | 100 (34.2-100) | 93.3 (70.2-98.8) | 66.7 (19.9-98.8) | 100 (89.5-100) |
| 4 | CVS | 41 | 7 | 1 | 33 | 0 | 17.1 | 100 (64.6-100) | 97.1 (85.1-99.5) | 87.5 (57.3-99.6) | 100 (92.2-100) |
| 4 | ES | 41 | 7 | 0 | 34 | 0 | 17.1 | 100 (64.6-100) | 100 (89.8-100) | 100 (66.7-100) | 100 (92.2-100) |
| 4 | PCyt | 43 | 7 | 1 | 35 | 0 | 16.3 | 100 (64.6-100) | 97.2 (85.8-99.5) | 87.5 (57.2-99.6) | 100 (92.6-100) |
| 5 | CVS | 145 | 1 | 0 | 144 | 0 | 0.7 | 100 (20.7-100) | 100 (97.4-100) | 100 (6.4-100) | 100 (99.3-100) |
| 5 | ES | 162 | 1 | 0 | 161 | 0 | 0.6 | 100 (20.7-100) | 100 (97.7-100) | 100 (6.4-100) | 100 (99.4-100) |
| 5 | PCyt | 167 | 1 | 0 | 166 | 0 | 0.6 | 100 (20.7-100) | 100 (97.7-100) | 100 (6.4-100) | 100 (99.4-100) |
| 6 | CVS | 67 | 10 | 2 | 55 | 0 | 14.9 | 100 (72.2-100) | 96.5 (89.1-99.0) | 83.3 (59.2-98.2) | 100 (94.8-100) |
| 6 | ES | 80 | 13 | 4 | 63 | 0 | 18.3 | 100 (77.2-100) | 94.0 (85.6-97.7) | 76.5 (57.1-92.2) | 100 (95.3-100) |
| 6 | PCyt | 92 | 20 | 3 | 69 | 0 | 21.7 | 100 (83.9-100) | 95.8 (88.5-98.6) | 87.0 (70.4-97.0) | 100 (95.5-100) |
| 7 | CVS | 173 | 18 | 3 | 152 | 0 | 10.4 | 100 (82.4-100) | 98.1 (94.5-99.3) | 85.7 (67.7-96.7) | 100 (97.9-100) |
| 7 | ES | 161 | 12 | 3 | 146 | 0 | 7.5 | 100 (75.8-100) | 98.0 (94.2-99.3) | 80.0 (58.3-95.4) | 100 (97.9-100) |
| 7 | PCyt | 194 | 18 | 4 | 172 | 0 | 9.3 | 100 (82.4-100) | 97.7 (94.3-99.1) | 81.8 (64.1-94.3) | 100 (98.1-100) |
| 8 | CVS | 80 | 10 | 2 | 68 | 0 | 12.5 | 100 (72.2-100) | 97.1 (90.2-99.2) | 83.3 (59.0-98.2) | 100 (95.8-100) |
| 8 | ES | 83 | 9 | 2 | 72 | 0 | 10.8 | 100 (70.1-100) | 97.3 (90.7-99.3) | 81.8 (56.3-97.4) | 100 (96.1-100) |
| 8 | PCyt | 88 | 9 | 0 | 77 | 0 | 10.5 | 100 (70.1-100) | 100 (95.2-100) | 100 (71.4-100) | 100 (96.2-100) |
| 9 | CVS | 45 | 10 | 1 | 34 | 0 | 22.2 | 100 (72.2-100) | 97.1 (85.5-99.5) | 90.9 (65.7-99.7) | 100 (91.9-100) |
| 9 | ES | 48 | 10 | 0 | 38 | 0 | 20.8 | 100 (72.2-100) | 100 (90.8-100) | 100 (74.0-100) | 100 (92.5-100) |
| 9 | PCyt | 48 | 10 | 1 | 37 | 0 | 20.8 | 100 (72.2-100) | 97.4 (86.5-99.5) | 90.9 (65.6-99.7) | 100 (92.5-100) |

Cl = confidence interval, CVS = cirician-collected vaginal swab, ES = endocervical swab, FN = false positive, PCyt = PreservCyt Solution liquid Pap. Prev = prevalence. TN = true negative, TP = true positive.

'Score confidence interval.

PPV 95% confidence interval computed from the exact 85% confidence interval for the positive likelihood ratio. NPV 95% confidence interval computed from the exact 95% confidence interval from the negative likelihood ratio

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Table 4 shows the sensitivity, specificity, PPV, and NPV of the APTIMA Trichomonas vaginalis Assay on the PANTHER System and the prevalence of 7. vaginalis (based on the infected status) in PreservCyt Solution liquid Pap specimens by cervical collection device. For PreservCyt Solution liquid Pap specimens, performance was similar across collection devices.

Table 4: Performance Characteristics of the APTIMA Trichomonas vaginalis Assay in PreservCyt Solution Liquid Pap Specimens by Collection Device Type

| Collection Device | n | TP | FP | TN | FN | Prev % | Sensitivity<br>(95% CI)1 | Specificity<br>(95% CI)1 | PPV %<br>(95% CI)2 | NPV %<br>(95% CI)2 |
|-------------------|-----|----|----|-----|----|--------|--------------------------|--------------------------|--------------------|--------------------|
| Broom-type Device | 414 | 54 | 5 | 355 | 0 | 13.0 | 100 (93.4-100) | 98.6 (96.8-99.4) | 91.5 (82.4-97.1) | 100 (99.0-100) |
| Spatula/Cytobrush | 360 | 29 | 5 | 326 | 0 | 8.1 | 100 (88.3-100) | 98.5 (96.5-99.4) | 85.3 (71.5-94.7) | 100 (99.0-100) |

Cl = confidence interval, FN = false positive, Prev = prevalence, TN = true negative, TP = true positive, Score confidence interval.

f PPV 95% confidence interval computed from the exact 95% confidence interval for the positive likelihood ratio, NPV 95% confidence interval computed from the exact 95% confidence interval from the negative likelihood ratio.

## Agreement of APTIMA Trichomas vaginalis Assay Results on the PANTHER System and the TIGRIS DTS System

It is recognized that device performance in an asymptomatic population is essential since the majority of individuals infected with Trichomonas vaginalis do not have symptoms. To further characterize performance of the assay in asymptomatic subjects, agreement between APTIMA Trichomonas vaginalis Assay results on the PANTHER System and the TIGRIS DTS System was assessed using prospectively collected specimens from asymptomatic subjects. Women were enrolled from 6 US clinical sites, including obstetrics and gynecology, family planning, and STD clinics. One (1) vaginal swab, 1 endocervical swab, and 1 PreservCyt Solution liquid Pap specimen were collected from each subject. All specimens were clinician-collected. PreservCyt liquid Pap specimens were collected with a broom-type device or a spatula and cytobrush.

APTIMA Trichomonas vaginalis Assay testing was conducted in accordance with the package insert instructions. PANTHER System testing was conducted at 3 sites (2 external laboratories and Gen-Probe). TIGRIS DTS System testing was conducted at Gen-Probe.

Eighteen (18) APTIMA Trichomonas vaginalis Assay runs were initiated on the PANTHER System; all were valid. A total of 227 vaginal swab, 227 endocervical swab, and 227 PreservCyt Solution liquid Pap specimens were tested. Of these specimens, 1 vaginal swab specimen (0.4%, 1/227) had an initial invalid result due to hardware error. The specimen with an initial invalid result was retested and had a valid result.

Of the samples with final valid APTIMA Trichomonas vaginalis Assay results on the PANTHER System, 227 vaginal swab, 227 endocervical swab, and 226 PreservCyt Solution liquid Pap o specimens had valid, paired results on the TIGRIS DTS System.

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Table 5 shows positive and negative percent agreements of APTIMA Trichomonas vaginalis Assay results on the PANTHER System and the TIGRIS DTS System in each specimen type for asymptomatic subjects.

| Specimen<br>Type | n | TIGRIS +<br>PANTHER + | TIGRIS -<br>PANTHER + | TIGRIS -<br>PANTHER - | TIGRIS +<br>PANTHER - | TIGRIS<br>Positivity | % Positive<br>Agreement<br>(95% CI)2 | % Negative<br>Agreement<br>(95% CI)2 |
|------------------|-----|-----------------------|-----------------------|-----------------------|-----------------------|----------------------|--------------------------------------|--------------------------------------|
| CVS1 | 227 | 29 | 5 | 191 | 2 | 13.7 | 93.5 (79.3-98.2) | 97.4 (94.2-98.9) |
| ES | 227 | 28 | 1 | 198 | 0 | 12.3 | 100 (87.9-100) | 99.5 (97.2-99.9) |
| PCyt | 228 | 26 | 1 | 199 | 0 | 11.5 | 100 (87.1-100) | 99.5 (97.2-99.9) |

| Table 5: Agreement between APTIMA Trichomonas vaginalis Assay Results on the | | |
|------------------------------------------------------------------------------|--|--|
| PANTHER System and the TIGRIS DTS System in Asymptomatic Subjects | | |

positive, - = negative, Cl = comfidence interval, CVS = clinician-collected vaginal swab, ES = PreservCyt Solution liquid Pap.

The 2 vaginal swab samples with positive APTIMA Trichomonas vaginalis Assay results on the TIGRIS DTS System and negative results on the PANTHER System were from subjects whose other samples had negative results on both the PANTHER System and the TIGRIS DTS System.

2 Score confidence interval.

### b.) Clinical Reproducibility:

The ATV Assay (Version 2) testing was conducted at 3 sites (in-house and 2 external) on the PANTHER System. Six operators, 2 at each site, performed reproducibility testing using 2 reagent lots over a period of at least 6 days. Four reproducibility panel members were prepared using a representative specimen matrix. The panels consisted of Trichomonas vaginalis negative and positive specimens prepared in PreservCyt solution. The panels included negative, high negative, moderate positive, and high positive samples. Identical panels were tested on 1 PANTHER System at each test site. Each run contained 3 replicates of each reproducibility panel member (12 PreservCyt samples total). Results are shown in the chart below (Table 6).

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| Conc | Target<br>Conc<br>(TV/mL) | N | Agmt<br>(%) | Mean<br>RLU | Between Sites | | Between<br>Operators | | Between Lots | | Between Runs | | Within Runs | | Totals | |
|------|---------------------------|-----|-------------|-------------|---------------|--------|----------------------|--------|--------------|--------|--------------|--------|-------------|--------|--------|--------|
| | | | | | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) |
| Neg | N/A | 108 | 99.1 | 23.5 | 0.0 | 0.0 | 2.7 | 11.6 | 0.0 | 0.0 | 0.0 | 0.0 | 37.5 | 159.7 | 37.6 | 160.1 |
| HNeg | 0.003 | 108 | 90.7 | 69.3 | 5.0 | 7.3 | 4.5 | 6.5 | 6.1 | 8.8 | 14.8 | 21.4 | 16.0 | 23.1 | 23.6 | 34.1 |
| MPos | 0.02 | 108 | 97.2 | 348.1 | 30.3 | 8.7 | 33.1 | 9.5 | 33.1 | 9.5 | 77.0 | 22.1 | 62.9 | 18.1 | 114.0 | 32.8 |
| HPos | 1.00 | 108 | 100 | 1185.5 | 0.0 | 0.0 | 17.0 | 1.4 | 0.0 | 0.0 | 28.0 | 2.4 | 34.2 | 2.9 | 47.4 | 4.0 |

## Table 6 - APTIMA Trichomas vaginalis Assay Reproducibility Study

Agmt = agreement, Conc = concentration, CV = coefficient of variation, HNeg = high negative, HPos = high positive,

MPos = moderate positive, Neg = negative, RLU = relative light units, SD = standard deviation.

Note: The RLU value reported by the software is the total measured RLU divided by 1000 with the decimal point truncated. Variability from some factors may have been numerically negative. This occurred if the variability due to those factors was very small. In these cases, SD and CV are shown as 0.

## c. Expected Values and Reference Range

The prevalence of T. vaginalis in different populations depends on patient risk factors such as age, lifestyle, the presence or absence of symptoms, and the sensitivity of the test in detecting the infection.

The positivity rate for the ATV Assay (Version 2) by specimen type, collection site and overall was 11.8% (80/678) for vaginal swabs, 11.2% (80/713) for endocervical swabs and 11.8% (93/790) for PreservCyt specimens.

## N. Instrument Name

The PANTHER System

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## O. System Descriptions:

- Modes of Operation a. Batch, random access
- b. Specimen Identification

By handheld barcode reader and positional checks

- Specimen Sampling and Handling C.
Fully Automated

- d. Calibration
Gen-Probe Field Service Engineers perform a luminometer calibration on the PANTHER System every 12 months as part of the Preventive Maintenance. Also. there are process controls and calibration checks on all of the dispenses, thermal devices, and the vacuum system.

- e. Quality Control
In addition to the assay controls that are specific to each assay, the PANTHER System contains process controls that employ both hardware and software components. The process controls include, but are not limited to:

- Verification that the sequence of assay processing steps is correct for each . reaction.
- . Verification that the reaction incubation times and temperatures are correct.
- . Verification that reagents and fluids were appropriately dispensed.

## P. Conclusion

The submitted information in this premarket notification demonstrates that the APTIMA Trichomonas vaginalis Assay on the PANTHER System is substantially equivalent to the predicate device.

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Image /page/16/Picture/0 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the perimeter. Inside the circle is an abstract symbol resembling an eagle or bird in flight, composed of three curved lines.

## DEPARTMENT OF HEALTH & HUMAN SERVICES

#### Public Health Service

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-002 .

## JAN 0-9 2013

Gen-Probe Incorporated C/O Maria Carmelita S. Baluyot 10210 Genetic Center Drive San Diego, CA 92121-4362

Re: k122062

Trade/Device Name: APTIMA® Trichomonas vaginalis Assay (PANTHER® System) Regulation Number: 21 CFR 866.3860 Regulation Name: Trichomonas vaginalis Nucleic Acid Amplification Test System Regulatory Class: Class II Product Code: OUY Dated: December 27, 2012 Received: December 28, 2012

Dear Ms. Baluyot:

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.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), 2 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 Part 801); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set

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

## Page 2 - Maria Carmelita S. Baluyot

forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

If you desire specific advice for your device on our labeling regulation (21 CFR Parts 801 and 809), please contact the Office of In Vitro Diagnostics and Radiological Health at [phone]. 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 Small Manufacturers, International and Consumer Assistance at its toll-free number [phone] or [phone] or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html.

Sincerely yours,

# Sally A. Hojvat

Sally A. Hojvat, MSc., PhD. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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## Indications for Use Form

## 510(k) Number (if known): K122062

Device Name: APTIMA® Trichomonas vaginalis Assay (PANTHER® System) Indications for Use:

The APTIMA Trichomonas vaginalis Assay is an in vitro qualitative nucleic acid amplification test (NAAT) for the detection of ribosomal RNA (rRNA) from Trichomonas vaginalis to aid in the diagnosis of trichomoniasis using the PANTHER System.

The assay may be used to test the following specimens from symptomatic or asymptomatic women: clinician-collected endocervical swabs, clinician-collected vaginal swabs, and specimens collected in PreservCyt Solution.

Prescription Use X (Part 21 CFR 801 Subpart D)

AND/OR

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

(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE OF NEEDED)

Concurrence of CDRH, Office of In Vitro Diagnostic Devices (OIVD)

Tamara Veldblbly

Division Sign-Off Office of In Vitro Diagnostic Device Evaluation and Safety

510(k) K122062

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