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 TIGRIS DTS System.

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

Device Description

The ATV assay is a nucleic acid amplification test intended for the in vitro qualitative detection of ribosomal RNA from T. vaginalis in patient-collected first catch urine and clinician collected vaginal swabs, endocervical swab and ThinPrep Pap Test specimens collected in Cytyc Preservcyt solution. The assay may be used to test specimens from symptomatic and asymptomatic individuals to aid in the diagnosis of trichomoniasis using the TIGRIS DTS System automated analyzer.

There are 4 kits (1 master and 3 ancillary) that are required to perform the ATV assay on the TIGRIS DTS System. The Master Kit contains 9 reagents and 2 controls and is made up of 3 boxes. Box 1 - the Refrigerated box contains ATV amplification reagent, ATV enzyme reagent, ATV probe reagent and ATV Target Capture reagent-B. Box 2 - the Room Temperature box contains ATV amplification reconstitution solution, ATV enzyme reconstitution reagent, ATV probe reconstitution reagent, ATV selection reagent and ATV target capture reagent. Box 3-the Controls kit box contains ATV positive and negative controls. The three ancillary kits consist of the APTIMA Assay Fluids kit, the APTIMA Auto Detect Reagents kit and APTIMA System Fluids Preservative kit. In addition to the reagents provided in the kit, the assay utilizes four specimen collection kits - the APTIMA unisex swab specimen collection kit for endocervical and male urethral swab specimens, APTIMA vaginal swab specimen collection kit, APTIMA urine specimen collection kit for male and female urine specimens and the APTIMA specimen transfer kit.

AI/ML Overview

Here's an analysis of the provided text to extract the acceptance criteria and study details for the Aptima Trichomonas vaginalis (ATV) assay.

Acceptance Criteria and Device Performance for Aptima Trichomonas vaginalis (ATV) Assay

The Aptima Trichomonas vaginalis (ATV) assay is a qualitative nucleic acid amplification test (NAAT) designed for the detection of ribosomal RNA (rRNA) from T. vaginalis. Its performance was evaluated through various analytical and clinical studies.

1. Acceptance Criteria and Reported Device Performance

The acceptance criteria for the ATV assay are implicitly defined by the reported performance characteristics which are deemed sufficient for reclassification to Class II. The primary performance metrics are related to the accuracy of T. vaginalis detection across different specimen types.

Metric	Acceptance Criteria (Implied)	Reported Device Performance
Clinical Sensitivity	High sensitivity across all claimed specimen types.	Urine: 95.2% (95% CI: 88.4-98.1) Clinician-collected vaginal swab: 100% (95% CI: 96.7-100) Endocervical swab: 100% (95% CI: 96.7-100) PreservCyt solution liquid Pap: 100% (95% CI: 96.0-100) Similar performance in symptomatic and asymptomatic women.
Clinical Specificity	High specificity across all claimed specimen types.	Urine: 98.9% (95% CI: 97.8-99.5) Clinician-collected vaginal swab: 99.0% (95% CI: 97.9-99.5) Endocervical swab: 99.4% (95% CI: 98.6-99.7) PreservCyt solution liquid Pap: 99.6% (95% CI: 98.8-99.9) Similar performance in symptomatic and asymptomatic women.
Positive Predictive Value (PPV)	High PPV, especially important for positive results.	Urine: 92.0% (95% CI: 1-96.4) Clinician-collected vaginal swab: 93.3% (95% CI: 87.6-97.0) Endocervical swab: 95.8% (95% CI: 90.7-98.6) PreservCyt solution liquid Pap: 96.9% (95% CI: 91.4-99.3)
Negative Predictive Value (NPV)	High NPV, important for ruling out infection.	Urine: 99.4% (95% CI: 98.5-99.8) Clinician-collected vaginal swab: 100% (95% CI: 99.5-100) Endocervical swab: 100% (95% CI: 99.6-100) PreservCyt solution liquid Pap: 100% (95% CI: 99.5-100)
Detection Limit	100% positivity at low T. vaginalis concentrations.	100% positivity for T. vaginalis at 0.1 TV/mL in urine, PreservCyt, and vaginal swab matrices for two T. vaginalis strains (Metronidazole-susceptible and Metronidazole-resistant).
Analytical Specificity	No significant cross-reactivity with common genitourinary flora or closely related organisms; minimal interference from other substances.	No cross-reactivity or significant effect on specificity with a wide range of microorganisms (Table 7 in the source document). No significant interference with most tested substances (e.g., lubricants, spermicides, anti-fungal/anti-itch medications, hormones, blood, urine controls) except for porcine gastric mucus (lower signal output). Lower signal outputs observed in the presence of Trichomonas tenax and Pentatrichomonas hominis.
Precision/Reproducibility	Consistent results from repeated testing across sites, operators, and reagent lots.	Coefficient of Variation (CV) for RLU values ranged from 4.4% to 74.1% across various panel members (high negative, moderate positive, high positive) and matrices (PreservCyt, Urine). Total CV for high positive samples was 14.1% (P) and 17.9% (U).
Assay Cut-off	Clear rules for test interpretation (Negative, Positive, Invalid).	Negative: Total RLU (x 1000) of 0* to <100 (where 0 indicates RLU 0-999 and valid for 690-999). Positive: Total RLU (x 1000) of 100 to <2400. Invalid: Total RLU (x 1000) of 0* (for RLU <690) or >/= 2400.
Control Acceptability	Controls must perform within specified RLU ranges.	Negative Control: Total RLU (x 1000) of 0* and <20 (Negative). Positive Control: Total RLU (x 1000) of >=500 and < 2400 (Positive).

2. Sample Size and Data Provenance for the Test Set (Clinical Study)

Sample Size: 1025 symptomatic and asymptomatic women were enrolled.
- Evaluable specimens:
  - Urine: 738 (3 had final invalid results)
  - Vaginal Swabs: 877 (2 had final invalid results)
  - Endocervical Swabs: 922 (2 had final invalid results)
  - PreservCyt solution liquid Pap specimens: 813
Data Provenance: The study was a "pivotal prospective multicenter clinical trial" conducted with women enrolled from 9 US clinical sites, including obstetric and gynecology, family planning, and STD clinics. This indicates prospective data collection from a diverse US population.

3. Number of Experts and Qualifications for Ground Truth

The document does not explicitly state the number or specific qualifications of individuals who interpreted the predicate tests to establish the ground truth. However, it indicates:

"2 of the vaginal swab specimens were tested with a commercially available culture system and wet mount microscopic exam to establish infected status."
This suggests that the ground truth was established by laboratory professionals and/or clinicians skilled in performing and interpreting these traditional diagnostic methods. The document does not specify their level of experience (e.g., radiologist with 10 years of experience).

4. Adjudication Method for the Test Set

The ground truth was established using a "patient infected status algorithm":

At least one positive reference result (from culture and/or wet mount microscopic exam) established an "infected patient status."
Both reference tests were required to be negative to establish a "non-infected patient status."

This effectively acts as an adjudication, where discordant results between the two reference methods (culture and wet mount) are resolved by prioritizing a positive finding from either method for "infected" status, and requiring both to be negative for "non-infected" status.

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

No, a Multi-Reader Multi-Case (MRMC) comparative effectiveness study was not performed or described in this document. The study focuses on comparing the assay's performance against traditional diagnostic methods, not on human reader improvement with or without AI assistance.

6. Standalone (Algorithm Only Without Human-in-the-Loop) Performance

Yes, the study describes the standalone performance of the APTIMA Trichomonas vaginalis Assay. The assay is an in vitro diagnostic device, and its results are automatically interpreted by the TIGRIS DTS System APTIMA Trichomonas vaginalis Software. The reported sensitivity, specificity, PPV, and NPV are characteristics of the automated assay's performance alone, without human interpretation or intervention in the diagnostic call process beyond initial specimen collection and loading.

7. Type of Ground Truth Used

The ground truth used for the clinical study was based on a composite reference standard:

Expert Consensus (implied by method): "commercially available culture system and wet mount microscopic exam"
Algorithm-based Patient Infected Status: An algorithm designated a subject as "infected" if at least one of the two reference methods (culture or wet mount) was positive, and "non-infected" if both were negative. This combines established diagnostic methods to form a "gold standard" for the study.

8. Sample Size for the Training Set

The document does not specify a separate training set or its sample size. The description of the clinical trial refers to a "pivotal prospective multicenter clinical trial" where all collected specimens were used for performance evaluation. For in vitro diagnostic assays, developers typically use internal data for assay development and optimization (which can be considered the "training phase"), but this internal data is not usually described in the same detail as the independent clinical validation (the "test set" in AI/ML terminology).

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

Since a specific "training set" is not explicitly mentioned or detailed in the context of this 510(k) submission, the method for establishing its ground truth is also not provided. If an internal "training set" was used during assay development, it would have likely relied on similar established methods (culture, wet mount, or other validated laboratory techniques) to determine the T. vaginalis status of samples for optimizing the assay's parameters.

Summary

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510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY AND INSTRUMENT COMBINATION TEMPLATE

A. 510(k) Number:

K102911

B. Purpose for Submission:

Evaluation of automatic Class III designation for the Aptima Trichomonas vaginalis (ATV) assay

C. Measurand:

Ribosomal RNA from T. vaginalis

D. Type of Test:

Nucleic acid amplification test

E. Applicant:

Gen Probe Inc.

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

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H. Intended Use:

1. 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 TIGRIS DTS System.

1. 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 TIGRIS DTS System.

1. Special conditions for use statement:
  For prescription use
1. Special instrument requirements:
  The automated TIGRIS DTS System

I. Device Description:

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vaginal swab specimen collection kit, APTIMA urine specimen collection kit for male and female urine specimens and the APTIMA specimen transfer kit.

J. Substantial Equivalence Information:

1. Predicate device name: In Pouch Trichomonas vaginalis
1. Predicate 510(k) number: K896296
1. Comparison with predicate:

Similarities
Item	Device	Predicate
Intended Use	NAAT test for detection ofT. vaginalis ribosomal RNA(rRNA)	Accurate system for earlymicroscopic identificationand culture confirmation ofT. vaginalis from femaleand male urogenital sites

Differences
Item	Device	Predicate
Specimen types	Urine, vaginal swab,endocervical swab, ThinPrep in PreservCyt solution	Vaginal swab ,seminalfluid, urine
Assay type	Target capture, transcriptionmediated amplificationassay	Culture media
Detection	Direct microscopicobservation, theninoculation for culture	Hybridization protectionassay, providing relativelight units (RLUs)

K. Standard/Guidance Document Referenced (if applicable):

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

EP 15-A2, 2006: User Verification of Performance For Precision and Trueness, CLSI approved Guideline

Format for Traditional and Abbreviated 510(k)s; Guidance for Industry and Staff, Aug.2005

General Principles of Software Validation; final guidance for Industry and FDA Staff, Jan. 2002

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Guidance for the Content of Premarket Submissions for Software contained in Medical Devices, May 2005

L. Test Principle:

The Aptima TV 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 (if/when applicable):

1. Analytical performance:
- a. Precision/Reproducibility:

Reproducibility was evaluated at 3 external US labs using the TIGRIS DTS System. Six operators, 2 at each site, performed reproducibility testing using 3 reagent kit lots. At each site, testing was performed over 6 days. Each site performed 2 runs per day. Each run contained 3 replicates of an 8 member reproducibility panel. The panels consisted of Trichomonas vaginalis negative and positive specimens prepared in either a urine or PreservCyt solution. For each sample matrix, there was a high negative, moderate positive, high positive and negative sample. Results are shown in the chart below.

						Between Sites			Between Operators			Between Lots			Between Worklists			Within Worklists
			Conc	Target		Mean		CV		CV		CV		CV		CV		CV
	PM	Matrix	Level	Conc¹	N	RLU	SD	(%)	SD	(%)	SD	(%)	SD	(%)	SD	(%)	SD	(%)
A	P	Neg	N/A	106	2.0	1.1	56.8	0.0	0.0	0.0	0.0	0.4	21.3	0.8	42.5	1.5	74.1
B	P	HNeg	0.01	106	58.3	17.2	29.4	0.0	0.0	11.1	19.1	0.0	0.0	22.2	38.0	30.2	51.7
C	P	MPos	0.1	108	367.0	32.8	8.9	0.0	0.0	57.5	15.7	51.0	13.9	140.6	38.3	163.6	44.6
D	P	HPos	1	107	1110.4	53.9	4.9	0.0	0.0	109.6	9.9	60.9	5.5	77.1	6.9	156.8	14.1
E	U	Neg	N/A	108	2.1	1.0	45.7	0.0	0.0	0.0	0.0	0.0	0.0	1.3	62.4	1.7	77.3
F	U	HNeg	0.006	107	60.2	11.2	18.7	0.0	0.0	9.6	15.9	9.8	16.2	12.0	19.9	21.4	35.6
G	U	MPos	0.1	107	781.6	53.2	6.8	0.0	0.0	66.6	8.5	56.0	7.2	83.7	10.7	131.9	16.9
H	U	HPos	1	108	1122.8	49.5	4.4	15.0	1.3	119.3	10.6	109.2	9.7	106.9	9.5	200.7	17.9

Reproducibility Study: Signal Variability of the ATV Assay by Panel Member, Including Samples With Discordant Test Results

Conc=concentration, HNeg=high positive, MPos=moderate positive, Neg=negative, P=PreservCyt, PM=panel member, U=Urine

1 Concentration units = trichomonads/mL

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 be numerically negative. This can occur if the variability due to those factors is very small. In these cases, SD and CV are shown as 0.

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b. Linearity/assay reportable range:

The ATV Assay is designed for and validated on the TIGRIS DTS System. The assay test results are automatically interpreted by the TIGRIS DTS System APTIMA Trichomonas vaginalis Software. A test result may be negative, positive or invalid as determined by the total 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 TIGRIS DTS 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.

Traceability, Stability, Expected values (controls, calibrators, or methods): C.
Data to support the recommended shipping and storage conditions for the vaginal swab, PreservCyt liquid Pap and urine specimens were generated with negative clinical specimens spiked with T. vaginalis to a final concentration of 250 TV/mL. Greater than 95% positivity was observed in all matrices (vaginal swab, PreservCyt liquid Pap, and urine) 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 trichomonads rRNA.

Control	Total RLU (x1000)	Trichomonasvaginalis Result
Negative Control	0* and <20	Negative
Positive Control	>/=500 and < 2400	Positive

The APTIMA Trichomonas vaginalis Controls must produce the following test results:

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d. Detection limit:

Panel samples containing 0.1 TV/mL in urine specimen matrix, PreservCyt liquid Pap specimen matrix, and vaginal swab matrix (90 replicates per matrix) were prepared with two strains of T. vaginalis (one Metronidazole-susceptible strain and one Metronidazoleresistant strain). Testing showed 100% positivity in all specimen matrices and in both T. vaginalis strains.

Analytical specificity: e.

Analytical specificity of the APTIMA Trichomonas vaginalis Assay was evaluated by testing various microorganisms, including common flora of the female genitourinary tract, opportunistic organisms, and closely related organisms. Testing was conducted in vaginal swab, PreservCyt liquid Pap, and urine matrices with 25 replicates of each isolate per matrix. The list of organisms and the concentrations tested are provided in Table 7. No cross-reactivity or significant effect on APTIMA Trichomonas vaginalis Assay specificity was observed with any of the organisms tested.

The APTIMA Trichomonas vaginalis Assay was also evaluated by testing the same organisms (Table 7) in vaginal swab, PreservCyt liquid Pap, and urine matrices spiked with T. vaginalis lysate to a final concentration of 2.5 TV/mL (25 replicates of each isolate per matrix). The APTIMA Trichomonas vaginalis 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 Pentatrichomonas hominis is a commensal of the large intestine.

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Microorganism	Concentration Tested
	STM	PreservCyt	Urine
Acinetobacter lwoffi	4.6x107 CFU/mL	4.6x107 CFU/mL	2.3x107 CFU/mL
Actinomyces israelii	2.1x108 CFU/mL	2.1x108 CFU/mL	1.1x108 CFU/mL
Atopobium vaginae	6.2x106 CFU/mL	6.2x106 CFU/mL	6.2x106 CFU/mL
Bacteroides fragilis	6.4x108 CFU/mL	6.4x108 CFU/mL	3.2x108 CFU/mL
Bifidobacterium adolescentis	7.2x107 CFU/mL	7.2x107 CFU/mL	3.6x107 CFU/mL
Campylobacter jejuni	7.2x107 CFU/mL	7.2x107 CFU/mL	3.6x107 CFU/mL
Candida albicans	1.2x108 CFU/mL	1.2x108 CFU/mL	5.9x107 CFU/mL
Candida glabrata	1.3x108 CFU/mL	1.4x108 CFU/mL	6.4x107 CFU/mL
Candida parapsilosis	9.2x107 CFU/mL	9.2x107 CFU/mL	4.6x107 CFU/mL
Candida tropicalis	1.8x107 CFU/mL	1.8x107 CFU/mL	9.1x106 CFU/mL
Chlamydia trachomatis	2.0x104 TCID50/mL	2.0x104 TCID 50/mL	2.0x104 TCID50/mL
Clostridium difficile	2.6x107 CFU/mL	2.6x107 CFU/mL	1.3x107 CFU/mL
Clostridium perfringens	1.9x108 CFU/mL	1.9x108 CFU/mL	9.4x107 CFU/mL
Corynebacterium genitalium	2.8x107 CFU/mL	2.8x107 CFU/mL	1.4x107 CFU/mL
Cryptococcus neoformans	5.8x107 CFU/mL	5.8x107 CFU/mL	2.9x107 CFU/mL
Enterobacter aerogenes	1.5x109 CFU/mL	1.5x109 CFU/mL	1.0x108 CFU/mL
Enterococcus faecalis	9.2x107 CFU/mL	9.2x107 CFU/mL	9.2x107 CFU/mL
Escherichia coli	2.2x108 CFU/mL	2.2x108 CFU/mL	2.2x108 CFU/mL
Fusobacterium nucleatum	1.3x108 CFU/mL	1.3x108 CFU/mL	6.4x107 CFU/mL
Gardnerella vaginalis	8.2x106 CFU/mL	8.2x106 CFU/mL	4.1x106 CFU/mL
Haemophilus ducreyi	2.1x109 CFU/mL	2.1x109 CFU/mL	3.1x109 CFU/mL
Herpes simplex virus I	2.0x105 TCID50/mL	2.0x105 TCID 50/mL	2.0x105 TCID50/mL
Herpes simplex virus II	2.0x105 TCID50/mL	2.0x105 TCID 50/mL	2.0x105 TCID50/mL
HIV-1	3.0x107 copies/mL	3.0x107 copies/mL	3.0x107 copies/mL
HPV 16 (SiHa)	1.0x105 cell/mL	1.0x105 cells/mL	1.0x105 cells/mL
Klebsiella oxytoca	9.6x108 CFU/mL	9.6x108 CFU/mL	4.8x108 CFU/mL
Lactobacillus acidophilus	1.0x108 CFU/mL	1.0x108 CFU/mL	5.2x107 CFU/mL
Lactobacillus jensenii	1.6x109 CFU/mL	1.6x109 CFU/mL	8.2x108 CFU/mL
Lactobacillus vaginalis	4.6x108 CFU/mL	4.6x108 CFU/mL	2.3x108 CFU/mL
Listeria monocytogenes	2.1x109 CFU/mL	2.1x109 CFU/mL	1.0x109 CFU/mL
Mobiluncus curtisii	4.1x107 CFU/mL	4.1x107 CFU/mL	4.1x107 CFU/mL
Mycoplasma hominis	1.0x108 CFU/mL	1.0x108 CFU/mL	1.0x108 CFU/mL
Neisseria gonorrhoeae	2.7x108 CFU/mL	2.7x108 CFU/mL	1.4x108 CFU/mL
Pentatrichomonas hominis	2.2x106 CFU/mL	2.2x106 CFU/mL	1.3x106 CFU/mL
Peptostreptococcus anaerobius	2.2x108 CFU/mL	2.2x108 CFU/mL	1.1x108 CFU/mL
Prevotella bivia	5.2x108 CFU/mL	5.2x108 CFU/mL	2.6x108 CFU/mL
Propionibacterium acnes	1.6x108 CFU/mL	1.6x108 CFU/mL	1.6x108 CFU/mL
Proteus mirabilis	1.2x109 CFU/mL	1.2x109 CFU/mL	6.0x108 CFU/mL
Pseudomonas aeruginosa	1.5x108 CFU/mL	1.5x108 CFU/mL	1.5x108 CFU/mL
Staphylococcus aureus	2.8x108 CFU/mL	2.8x108 CFU/mL	2.8x108 CFU/mL
Staphylococcus epidermidis	3.0x108 CFU/mL	3.0x108 CFU/mL	1.5x108 CFU/mL
Streptococcus pyogenes	1.0x108 CFU/mL	1.0x108 CFU/mL	8.9x107 CFU/mL
Streptococcus agalactiae	1.0x108 CFU/mL	1.0x108 CFU/mL	1.0x108 CFU/mL
Trichomonas tenax	2.7x105 CFU/mL	2.7x105 CFU/mL	1.3x105 CFU/mL
Ureaplasma urealyticum	1.6x108 CFU/mL	1.4x108 CFU/mL	1.3x108 CFU/mL

Table 7: Microorganisms Tested in the APTIMA Trichomonas vaginalis Assay

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Interference Studies

The following substances (at a concentration of 1% vol/vol or wt/vol) were individually spiked into vaginal swab, PreservCyt liquid Pap, and urine matrices and tested in the APTIMA Trichomonas vaginalis Assay: over-the-counter personal lubricants, spermicides, deodorant sprays/powders, anti-fungal/anti-itch medications, intravaginal hormones, porcine gastric mucus, glacial acetic acid, vinegar, and seminal fluid. Whole blood was tested at 10% vol/vol and KOVA-Trol I High Abnormal w/ Urobilinogen Urinalysis Control was substituted for urine to test for high levels of protein, glucose, ketones, bilirubin, nitrite, and urobilinogen. No interference was observed with any of the tested substances in the APTIMA Trichomonas vaginalis Assay with the exception of porcine gastric mucus, which exhibited lower signal output when present at a final concentration of 1% (V/V or W/V).

f. Assay cut-off:

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

*If the RLU measured on the TIGRIS DTS System in between 0 and 999, a result of "0" 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.

1. Comparison studies:
- a. Method comparison with predicate device:

See 3 (a) below

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b. Matrix comparison:

N/A

3. Clinical studies:

a. Clinical Sensitivity:
A pivotal prospective multicenter clinical trial was conducted with 1025 symptomatic and asymptomatic women enrolled from 9 US clinical sites, including obstetric and gynecology, family planning and STD clinics. Up to 6 specimens were collected from each subject (1 first catch urine, 3 vaginal swabs, 1 endocervical swab and 1 PreservCyt solution liquid Pap specimen). All specimens were clinician-collected except urine specimens. 2 of the vaginal swab specimens were tested with a commercially available culture system and wet mount microscopic exam to establish infected status. The remaining 4 specimens were tested with the ATV assay at 3 external labs. Performance characteristics of the ATV assay were determined by comparing results to a patient infected status algorithm. Each subject was designated as infected or non-infected based on vaginal swab specimen results tested by culture and/or wet mount microscopic exam. At least one positive reference result established an infected patient status. Both reference tests were required to be negative to establish a non-infected patient status. Of the evaluable specimens, a total of 738 urines, 877 vaginal swabs, 922 endocervical swabs and 813 PreservCyt solution liquid Pap specimens were tested with the assay. There were 3 urines, 2 vaginal and 2 endocervical swabs with final invalid results due to hardware errors or specimen issues. Results below show the sensitivity, specificity, positive value (PPV), and negative predictive value (NPV) of the APTIMA Trichomonas vaginalis Assay and the prevalence of T. vaginalis (based on the infected status) in each specimen type. Performance was similar across specimen types.

SpecimenType	N	TP	FP	TN	FN	Prev%	Sensitivity%(95% CI)	Specificity%(95%CI)	PPV%(95%CI)	NPV%(95%CI)
Urine	735	80	7	644	4	11.4	95.2 (88.4-98.1)	98.9 (97.8-99.5)	92.0 (1-96.4)	99.4(98.5-99.8)
Cliniciancollectedvaginal swab	875	111	8	756	0	12.7	100 (96.7-100)	99.0(97.9-99.5)	93.3(87.6-97.0)	100(99.5-100)
Endocervicalswab	920	114	5	801	0	12.4	100 (96.7-100)	99.4(98.6-99.7)	95.8(90.7-98.6)	100(99.6-100)

Performance characteristics of the APTIMA Trichomonas vaginalis Assay

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PreservCyt	813	93	3	717	0	11.4	100 (96.0-100)	99.6 (98.8-99.9)	96.9(91.4-99.3)	100(99.5-100)
------------	-----	----	---	-----	---	------	--------------------	----------------------	---------------------	-------------------

The sensitivity, specificity, PPV, and NPV of the APTIMA Trichomonas vaginalis Assay and the prevalence of T. vaginalis (based on the infected status) in each specimen type were also evaluated by symptom status. Subjects were classified as symptomatic if symptoms were reported by the subject. Subjects were classified as asymptomatic if the subject did not report symptoms. For each specimen type, performance was similar in symptomatic and asymptomatic women. Prevalence was higher in symptomatic women.

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SpecimenType	SymptomStatus	n	TP	FP	TN	FN	Prev %	Sensitivity %(95% CI)¹	Specificity %(95% CI)¹	PPV %(95% CI)²	NPV %(95% CI)²
Urine	Asymptomatic	324	21	3	299	1	6.8	95.5(78.2-99.2)	99.0(97.1-99.7)	87.5(71.4-96.9)	99.7(98.4-100)
	Symptomatic	411	59	4	345	3	15.1	95.2(86.7-98.3)	98.9(97.1-99.6)	93.7(85.7-98.1)	99.1(97.7-99.8)
CVS	Asymptomatic	345	24	4	317	0	7.0	100(86.2-100)	98.8(96.8-99.5)	85.7(70.3-95.6)	100(98.9-100)
	Symptomatic	530	87	4	439	0	16.4	100(95.8-100)	99.1(97.7-99.6)	95.6(89.5-98.8)	100(99.2-100)
ES	Asymptomatic	372	26	1	345	0	7.0	100(87.1-100)	99.7(98.4-99.9)	96.3(82.4-99.9)	100(99.0-100)
	Symptomatic	548	88	4	456	0	16.1	100(95.8-100)	99.1(97.8-99.7)	95.7(89.6-98.8)	100(99.2-100)
PCyt	Asymptomatic	353	23	0	330	0	6.5	100(85.7-100)	100(98.8-100)	100(86.2-NC)	100(99.0-100)
	Symptomatic	460	70	3	387	0	15.2	100(94.8-100)	99.2(97.8-99.7)	95.9(88.9-99.1)	100(99.1-100)

Table 3: Performance Characteristics of the APTIMA Trichomonas vaginalis Assay by Symptom Status

CI = confidence interval, CVS = clinician-collected vaginal swab, ES = endocervical swab, FN = false negative, FP = false positive, NC = not calculable, PCyt = PreservCyt Solution liquid Pap, Prev = prevalence, TN = true negative, TP = true positive.

4Score confidence interval.

2PPV 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. Some confidence limits could not be calculated due to undefined results in the formulas.

Clinical specificity: b.
See 3(a) above
Other clinical supportive data (when a. and b. are not applicable): C.
N/A
1. Clinical cut-off:
  N/A
1. Expected values/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. A summary of the prevalence of T. vaginalis, by specimen type, as determined by the APTIMA Trichomonas vaginalis Assay in the clinical trial is described below.

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The positivity rate for the Aptima TV assay by specimen type, collection site and overall was 11.8% (87/735) for urines, 11.8% (96/813) for PreservCyt solution Pap specimens, 12.9% (119/920) for endocervical swab specimens and 13.6% (119/875) for vaginal swab specimens.

N. Instrument Name:

The TIGRIS DTS System

O. System Descriptions:

1. Modes of Operation:
  The TIGRIS DTS System is an integrated hardware and software system that fully automates all steps of nucleic acid testing necessary to perform Gen-Probe assays. The system automates the following steps: sample processing/ target capture, amplification, detection and results processing. The 2 main components of the system are the computer work station and the analyzer. The assay software in the computer work station directs the analyzer modules to perform each sequential assay step. The analyzer holds all of the fluids, reagents and consumables needed to perform the assay.
1. Software:
  FDA has reviewed applicant's Hazard Analysis and software development processes for this line of product types:

Yes _X _________ or No _______________________________________________________________________________________________________________________________________________________

1. Specimen Identification:
  See Section L, Test Principle. A complete description is included in theTIGRIS DTS System Operator's Manual for system procedural information
1. Specimen Sampling and Handling:
  See #3 above
1. Calibration:
  N/A
1. Quality Control:
  See #3 above

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P. Other Supportive Instrument Performance Characteristics Data Not Covered In The "Performance Characteristics" Section above:

N/A

Q. Proposed Labeling:

The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.

R. Conclusion:

The submitted information in this premarket notification is complete and supports reclassification into Class II

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.