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 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:

Refrigerated Box Contains the Amplification Reagent, Enzyme Reagent, Probe Reagent and Target Capture Reagent-B
Room Temperature Box Contains Amplification Reconstitution Solution, Enzyme Reconstitution Solution, Probe Reconstitution Solution, Selection Reagent and Target Capture Reagent
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.

APTIMA Unisex Swab Specimen Collection Kit for Endocervical and Male Urethral Swab Specimens
APTIMA Vaginal Swab Specimen Collection Kit
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).

AI/ML Overview

Acceptance Criteria and Device Performance Study for APTIMA® Trichomonas vaginalis Assay (PANTHER® System)

This section provides a summary of the acceptance criteria and the study conducted to demonstrate the APTIMA® Trichomonas vaginalis Assay on the PANTHER® System meets these criteria.

1. Table of Acceptance Criteria and Reported Device Performance

The primary acceptance criteria for this diagnostic device are its clinical sensitivity and specificity across different specimen types and symptom statuses.

Performance Metric	Specimen Type	Symptom Status	Acceptance Criteria (Implicit from prior clearance/predicate, demonstrated as 95% Confidence Interval)	Reported Device Performance (95% CI)
Sensitivity	Clinician-collected Vaginal Swab (CVS)	Asymptomatic	Expected to be high (e.g., >80% or 90%)	100% (75.8-100)
		Symptomatic	Expected to be high	100% (93.7-100)
		Overall	Expected to be high	100% (94.7-100)
	Endocervical Swab (ES)	Asymptomatic	Expected to be high	100% (80.6-100)
		Symptomatic	Expected to be high	100% (93.0-100)
		Overall	Expected to be high	100% (94.6-100)
	PreservCyt Solution liquid Pap (PCyt)	Asymptomatic	Expected to be high	100% (83.2-100)
		Symptomatic	Expected to be high	100% (94.3-100)
		Overall	Expected to be high	100% (95.6-100)
Specificity	Clinician-collected Vaginal Swab (CVS)	Asymptomatic	Expected to be high (e.g., >95%)	97.3% (94.6-98.7)
		Symptomatic	Expected to be high	98.8% (97.0-99.5)
		Overall	Expected to be high	98.2% (96.7-99.0)
	Endocervical Swab (ES)	Asymptomatic	Expected to be high	98.3% (96.1-99.3)
		Symptomatic	Expected to be high	97.9% (95.8-99.0)
		Overall	Expected to be high	98.1% (96.7-98.9)
	PreservCyt Solution liquid Pap (PCyt)	Asymptomatic	Expected to be high	99.4% (97.7-99.8)
		Symptomatic	Expected to be high	97.9% (95.9-98.9)
		Overall	Expected to be high	98.6% (97.4-99.2)

Note: The document does not explicitly state numerical acceptance criteria in a structured table. However, the reported performance characteristics (Sensitivity, Specificity, PPV, NPV) with narrow 95% Confidence Intervals consistently demonstrate high agreement with the "patient infected status algorithm," indicating that the device performs as expected for a diagnostic test of this nature, meeting an implicit acceptance threshold for high diagnostic accuracy. The agreement studies with the predicate device further support this.

2. Sample Sizes Used for the Test Set and Data Provenance

The clinical performance study used the following sample sizes for the test set:

Vaginal Swabs: 667 (after exclusions for invalid results which were 11 out of 689 initial samples)
Endocervical Swabs: 700 (after exclusions for invalid results which were 24 out of 737 initial samples)
PreservCyt Solution liquid Pap specimens: 774 (after exclusions for invalid results which were 1 out of 791 initial samples)

Data Provenance: The study utilized retrospective, leftover specimens collected from consenting subjects during a previous, prospective, multicenter clinical study of the ATV Assay on the TIGRIS DTS System.

Country of Origin: 9 US clinical sites (obstetrics and gynecology, family planning, and STD clinics).

For the agreement study with the TIGRIS DTS System for asymptomatic subjects:

Vaginal Swabs: 227
Endocervical Swabs: 227
PreservCyt Solution liquid Pap specimens: 226
Data Provenance: Prospectively collected specimens from asymptomatic subjects enrolled from 6 US clinical sites.

3. Number of Experts Used to Establish Ground Truth and Qualifications

The document does not specify the number of experts or their specific qualifications (e.g., years of experience for radiologists) for establishing the ground truth for the clinical study.

4. Adjudication Method for the Test Set

The ground truth for the clinical performance study was established by a patient infected status algorithm based on the results from two reference tests performed on vaginal swab specimens:

Commercially available culture system
Wet mount microscopic examination

Adjudication Rule:

Infected Patient Status: At least one of the reference test results was required to be positive.
Non-infected Patient Status: Both reference tests were required to be negative.

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study

No Multi-Reader Multi-Case (MRMC) comparative effectiveness study was done. This is a diagnostic assay (Nucleic Acid Amplification Test) and its performance is determined by its analytical and clinical characteristics against a defined ground truth, not by human reader interpretation. No human readers are involved in the direct interpretation of the assay results, which are automatically interpreted by the PANTHER System software.

6. Standalone Performance Study (Algorithm Only)

Yes, a standalone performance study (algorithm only, without human-in-the-loop performance) was done. The entire evaluation of the APTIMA® Trichomonas vaginalis Assay on the PANTHER® System, including its analytical and clinical performance, is based on the automated interpretation of the test results by the PANTHER System's software. The assay results (RLU values) are automatically interpreted as negative, positive, or invalid by the system.

7. Type of Ground Truth Used

The type of ground truth used for the clinical studies was an expert consensus-based algorithm derived from established diagnostic methods:

Culture for Trichomonas vaginalis
Wet mount microscopic examination

This algorithm defined the "patient infected status" against which the device's performance was measured.

8. Sample Size for the Training Set

The document does not explicitly state the sample size for a "training set" in the context of developing the algorithm itself. The information provided focuses on the validation data set used for clinical performance evaluation. Medical devices, especially diagnostic assays, often undergo development and internal validation on various sample sets, but these are typically not reported as explicitly as training sets in the context of machine learning model development. The focus here is on the analytical and clinical validation of the final assay.

9. How the Ground Truth for the Training Set was Established

As noted above, an explicit "training set" for the algorithm's development is not detailed. The ground truth for validating the assay's performance (the clinical test set) was established using a patient infected status algorithm based on a combination of culture and wet mount microscopic examination results. This is a common practice for validating new diagnostic tests against existing gold standards or established diagnostic pathways.

Summary

{0}------------------------------------------------

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 (858) 410-8000

Company Contact:	Ma. Carmelita S. BaluyotRegulatory Affairs Specialist
Phone:	858-410-8309
Fax:	858-410-9028
Email:	carmelita.baluyot@gen-probe.com
Date Prepared:	December 18, 2012

{1}------------------------------------------------

F. Proprietary and Established Names:

APTIMA Trichomonas vaginalis Assay

G. Regulatory Information

1. Regulation Section
21 CFR 866.3860
1. Classification Class II
1. Product Code OUY - Trichomonas vaginalis nucleic acid amplification test system
1. 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.

2. Indications for Use

1. Special conditions for use statement
  For prescription use

{2}------------------------------------------------

4. Special instrument requirements

The automated PANTHER System

I. Device Description

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

1. APTIMA Unisex Swab Specimen Collection Kit for Endocervical and Male Urethral Swab Specimens
1. APTIMA Vaginal Swab Specimen Collection Kit
1. 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).

{3}------------------------------------------------

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)Device	ATV Assay (K102911)Predicate
Device Class	2	same
Qualitative/Quantitative Assay	Qualitative	same
Intended Use	NAAT test for detection ofTrichomonas vaginalisribosomal RNA (rRNA)	same
Technology	Target Capture(TC)Transcription MediatedAmplification(TMA)Hybridization ProtectionAssay (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:• Vaginal swab• Endocervical swab• ThinPrep in PreservCyt solution	Four (4) Female specimen types:• Urine• Vaginal swab• Endocervical swab• ThinPrep in PreservCyt solution

{4}------------------------------------------------

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

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.

{5}------------------------------------------------

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	PanelMember	TV/mL	Valid N	%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.

{6}------------------------------------------------

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 vaginalisResult
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.

{7}------------------------------------------------

					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.

{8}------------------------------------------------

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.

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

2. Comparison Studies

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

{9}------------------------------------------------

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.

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

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.

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

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

{11}------------------------------------------------

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	SpecimenType	n	TP	FP	TN	Prev %	Sensitivity(95% CI)¹	Specificity(95% CI)¹	PPV %(95% CI)²	NPV %(95% CI)²
1	CVS	52	8	1	43	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	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	18.6	100 (74.1-100)	100 (92.6-100)	100 (75.6-100)	100 (93.9-100)
2	CVS	52	3	1	48	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	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	7.4	100 (58.6-100)	100 (94.3-100)	100 (58.3-100)	100 (96.0-100)
3	CVS	12	2	0	10	16.7	100 (34.2-100)	100 (72.2-100)	100 (32.1-100)	100 (85.6-100)
3	ES	16	2	0	14	12.5	100 (34.2-100)	100 (78.5-100)	100 (31.5-100)	100 (89.3-100)
3	PCyt	17	2	1	14	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	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	17.1	100 (64.6-100)	100 (89.8-100)	100 (66.7-100)	100 (92.2-100)
4	PCyt	43	7	1	35	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	0.7	100 (20.7-100)	100 (97.4-100)	100 (6.4-100)	100 (99.3-100)
5	ES	162	1	0	161	0.6	100 (20.7-100)	100 (97.7-100)	100 (6.4-100)	100 (99.4-100)
5	PCyt	167	1	0	166	0.6	100 (20.7-100)	100 (97.7-100)	100 (6.4-100)	100 (99.4-100)
6	CVS	67	10	2	55	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	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	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	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	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	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	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	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	10.5	100 (70.1-100)	100 (95.2-100)	100 (71.4-100)	100 (96.2-100)
9	CVS	45	10	1	34	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	20.8	100 (72.2-100)	100 (90.8-100)	100 (74.0-100)	100 (92.5-100)
9	PCyt	48	10	1	37	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

{12}------------------------------------------------

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(95% CI)1	Specificity(95% CI)1	PPV %(95% CI)2	NPV %(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.

{13}------------------------------------------------

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.

SpecimenType	n	TIGRIS +PANTHER +	TIGRIS -PANTHER +	TIGRIS -PANTHER -	TIGRIS +PANTHER -	TIGRISPositivity	% PositiveAgreement(95% CI)2	% NegativeAgreement(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).

{14}------------------------------------------------

Conc	TargetConc(TV/mL)	N	Agmt(%)	MeanRLU	Between Sites		BetweenOperators		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

{15}------------------------------------------------

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.

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

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 (301) 796-5450. 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 (800) 638-2041 or (301) 796-7100 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

{18}------------------------------------------------

Indications for Use Form

510(k) Number (if known): K122062

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

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

Page 1 of 1

Regulation Number and Section

§ 866.3860

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