GEN-PROBE APTIMA COMBO 2 ASSAY

K043224 · Gen-Probe, Inc. · LSL · Aug 9, 2005 · Microbiology

Device Facts

• Record ID: K043224
• Device Name: GEN-PROBE APTIMA COMBO 2 ASSAY
• Applicant: Gen-Probe, Inc.
• Product Code: LSL · Microbiology
• Decision Date: Aug 9, 2005
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3390
• Device Class: Class 2

Intended Use

The APTIMA Combo 2 Assay is a target amplification nucleic acid probe test that utilizes target capture for the in vitro qualitative detection and differentiation of ribosomal RNA (rRNA) from Chlamydia trachomatis (CT) and/or Neisseria gonorrhoeae (GC) in clinician-collected endocervical, vaginal, and male urethral swab specimens, patientcollected vaginal swab specimens*, and female and male urine specimens. The assay is also intended for use with testing of gynecological specimens collected in the PreservCyt Solution and processed with the Cytyc ThinPrep 2000 System. The assay may be used to test specimens from symptomatic and asymptomatic individuals to aid in the diagnosis of gonococcal and/or chlamydial urogenital disease. *Patient-collected vaginal swab specimens are an option for screening women when a pelvic exam is not otherwise indicated. The vaginal swab specimen collection kit is not for home use. The GEN-PROBE® APTIMA® Specimen Transfer Kit is only for use with GEN-PROBE APTIMA assays for the detection of Chlamydia trachomatis and/or Neisseria gonorrhoeae. The GEN-PROBE APTIMA Specimen Transfer Kit allows for APTIMA Assay testing of gynecological specimens collected and processed by the Cytyc ThinPrep 2000 Processor according to the instructions provided.

Device Story

The APTIMA Combo 2 Assay is a nucleic acid amplification test (NAAT) for detecting Chlamydia trachomatis and Neisseria gonorrhoeae. It utilizes target capture technology to isolate rRNA from clinical specimens. The assay is performed in a laboratory setting by trained technicians. The APTIMA Specimen Transfer Kit enables the use of gynecological specimens previously processed by the Cytyc ThinPrep 2000 System for Pap testing. The assay provides qualitative results, aiding clinicians in diagnosing urogenital infections. By enabling the use of existing liquid-based cytology samples, the device streamlines diagnostic workflows, allowing for concurrent screening for STIs and cervical abnormalities from a single specimen collection.

Clinical Evidence

Prospective multi-center study of 1,647 female subjects (symptomatic and asymptomatic). Sensitivity and specificity were calculated against a patient-infected status algorithm (requiring positive results from two reference NAATs). For C. trachomatis, overall sensitivity was 96.7% and specificity was 99.2%. For N. gonorrhoeae, overall sensitivity was 92.3% and specificity was 99.8%. Analytical studies confirmed no interference from common cervical substances and demonstrated stability of spiked samples under various storage conditions.

Technological Characteristics

Nucleic acid amplification test (NAAT) utilizing target capture and rRNA detection. Components include transport media with penetrable cap. Compatible with Cytyc ThinPrep 2000 Processor. Analytical sensitivity: 1 IFU/assay for C. trachomatis; 50 cells/assay for N. gonorrhoeae. Standalone laboratory assay.

Indications for Use

Indicated for qualitative detection and differentiation of CT and/or GC rRNA in symptomatic and asymptomatic individuals to aid in diagnosis of urogenital disease. Specimens include clinician-collected endocervical, vaginal, and male urethral swabs; patient-collected vaginal swabs; female and male urine; and gynecological specimens in PreservCyt Solution processed via ThinPrep 2000.

Regulatory Classification

Identification

Neisseria spp. direct serological test reagents are devices that consist of antigens and antisera used in serological tests to identify Neisseria spp. from cultured isolates. Additionally, some of these reagents consist of Neisseria spp. antisera conjugated with a fluorescent dye (immunofluorescent reagents) which may be used to detect the presence of Neisseria spp. directly from clinical specimens. The identification aids in the diagnosis of disease caused by bacteria belonging to the genus Neisseria, such as epidemic cerebrospinal meningitis, meningococcal disease, and gonorrhea, and also provides epidemiological information on diseases caused by these microorganisms. The device does not include products for the detection of gonorrhea in humans by indirect methods, such as detection of antibodies or of oxidase produced by gonococcal organisms.

Predicate Devices

• APTMA Combo 2 Assay (K003395)

Submission Summary (Full Text)

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# AUG 9 - 2005

# Summary of Safety and Effectiveness

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.

The assigned 510(k) number is: K043224

GEN-PROBE® APTIMA COMBO2® Assay GEN-PROBE® APTIMA® Specimen Transfer Kit

## Sponsor/Contact Information

Submitted By: Gen-Probe Incorporated 10210 Genetic Center Drive San Diego, California 92121 Phone: [phone]

Company Contact: Alan Maderazo, Ph.D., RAC Sr. Regulatory Affairs Specialist Phone: [phone] Fax: [phone] e-mail: [e-mail]

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## General Information

| Trade Name: | GEN-PROBE® APTIMA COMBO2® Assay |
|-----------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Common or Usual Name: | Ribosomal RNA (rRNA) target-amplified nucleic acid prob<br>test for the in vitro diagnostic detection of Chlamydia<br>trachomatis and/or Neisseria gonorrhoeae |
| Classification Names: | DNA Probe, Nucleic Acid Amplification, Chlamydia<br>DNA Reagents, Neisseria |

# APTIMA Combo 2 Assay

DNA Probe, Nucleic Acid Amplification, Chlamydia Device Description Medical Specialty Microbiology Product Code MKZ Device Class l Regulation number 866.3120

| Device Description | DNA Reagents, Neisseria |
|--------------------|-------------------------|
| Medical Specialty | Microbiology |
| Product Code | LSL |
| Device Class | 2 |
| Regulation number | 866.3390 |

### Substantially Equivalent Devices:

APTIMA Combo 2 Assay (K003395)

Roche COBAS AMPLICOR™ CT/NG Test (P950039/S008)

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### Device Description

Clearance of this premarket notification extends the clinical performance claims of the commercially available GEN-PROBE APTIMA Combo 2 Assay to include PreservCyt liquid Pap specimens (collected and processed by the Cytyc ThinPrep 2000 Processor) as acceptable testing specimens. The ancillary kit formulated for this specific application is the GEN-PROBE APTIMA Specimen Transfer Kit. The components of the APTIMA Specimen Transfer Kit include: (1) a transport tube containing transport media with a penetrable cap and (2) specific instructions for use regarding decontamination and specimen processing procedures. The APTIMA Specimen Transfer Kit may only be used in conjunction with GEN-PROBE APTIMA Assays for the detection of Chlamydia trachomatis and/or Neisseria gonorrhoeae..

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## Intended Use

### AC2 Assay package insert:

The APTIMA Combo 2 Assay is a target amplification nucleic acid probe test that utilizes target capture for the in vitro qualitative detection and differentiation of ribosomal RNA (rRNA) from Chlamydia trachomatis (CT) and/or Neisseria gonorrhoeae (GC) in clinician-collected endocervical, vaginal, and male urethral swab specimens, patientcollected vaginal swab specimens*, and female and male urine specimens. The assay is also intended for use with testing of gynecological specimens collected in the PreservCyt Solution and processed with the Cytyc ThinPrep 2000 System. The assay may be used to test specimens from symptomatic and asymptomatic individuals to aid in the diagnosis of gonococcal and/or chlamydial urogenital disease.

*Patient-collected vaginal swab specimens are an option for screening women when a pelvic exam is not otherwise indicated. The vaginal swab specimen collection kit is not for home use.

### Ancillary Kit package insert:

The GEN-PROBE® APTIMA® Specimen Transfer Kit is only for use with GEN-PROBE APTIMA assays for the detection of Chlamydia trachomatis and/or Neisseria gonorrhoeae. The GEN-PROBE APTIMA Specimen Transfer Kit allows for APTIMA Assay testing of gynecological specimens collected and processed by the Cytyc ThinPrep 2000 Processor according to the instructions provided.

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#### APTIMA Combo 2 Assay

A complete description of the APTIMA Combo 2 Assay is provided in the commercialized package insert.

## Summary of Non-Clinical (Analytical Laboratory) Performance Data

#### Limit of Detection (Analytical Sensitivity)

Chlamydia trachomatis analytical sensitivity (limits of detection) was determined by directly comparing dilutions of C. trachomatis organisms in cell culture and in the assay. The analytical sensitivity claim for the assay is one Inclusion-Forming Unit (IFU) per assay (9.75 IFU/mL PreservCyt liquid Pap) for all 15 C. trachomatis serovars. However, dilutions of less than 1 IFU/assay of all serovars tested positive in the APTIMA Combo 2 Assay.

Neisseria gonorrhoeae analytical sensitivity was determined by directly comparing dilutions of 20 different clinical isolates in culture and in the APTIMA Combo 2 Assay. The analytical sensitivity claim for the assay is 50 cells/assay (488 cells/mL PreservCyt liquid Pap). However, all strains tested were positive at less than 50 cells/assay.

#### Analytical Specificity

The Chlamydia and Neisseria species were used to evaluate the analytical specificity of the AC2 Assay. A total of 50 culture isolates were tested in the PreservCyt liquid Pap media. These isolates included 47 Neisseria strains and two species of Chlamydia (two strains of Chlamydia psittaci and one strain of Chlamydia pneumoniae). All organisms except C. psittaci and C. pneuomoniae were tested at greater than 1.0 X 106 cells/assay in the mixed solution (PreservCyt/STM). For C. psittaci, two isolates were tested - VR601 at a concentration of 7.91 x 104 cells/assay, and VR629 at a concentration of 1 x 10 CELDss/0.2 mL. For C. pneumoniae (VR1360), the tested concentration was 3.95 x 10

Gen-Probe Incorporated

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cells/assay. The list of organisms tested and their corresponding AC2 Assay results are provided in the Table below

| ORGANISM | ORGANISM |
|------------------------|---------------------------------|
| Chlamydia psittaci | Neisseria cinerea (4) |
| Chlamydia psittaci | Neisseria dentrificans |
| Chlamydia pneumoniae | Neisseria lactamica (9) |
| Neisseria elongate (3) | N. meningitidis Serogroup A |
| Neisseria flava | N. meningitidis Serogroup B |
| Neisseria mucosa | N. meningitidis Serogroup C (4) |
| Neisseria mucosa | N. meningitidis Serogroup D |
| Neisseria perflava | N. meningitidis Serogroup Y |
| Neisseria sicca | N. meningitidis Serogroup W135 |
| Neisseria sicca | Neisseria polysaccharea |
| Neisseria subflava(14) | |

Analytical Specificity - PreservCyt liquid Pap Specimen Study

(n) = number of strains tested

All organisms tested produced a negative result in the APTIMA Combo 2 Assay.

#### Interference Studies

The following substances commonly present in cervical specimens that were tested in the assay. 10% blood, contraceptive jelly, spermicide, moisturizer, hemorrhoidal anesthetic, body oil, powder, anti-fungal cream, vaginal lubricants, feminine spray, and leukocytes (1 x 106 cells/mL).

All were tested for potential assay interference in the absence and presence of C. trachomatis and N. gonorrhoeae at the estimated rRNA equivalent of one C. trachomatis IFU/assay (5 fg/assay) and 50 N. gonorrhoeae cell/assay (250 fg/assay). The rRNA equivalents were calculated based on the genome size and estimated DNA:RNA ratio/cell of each organism. No interference was observed with any of the tested substances.

#### Recovery

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Escherichia coli, Gardnerella vaginalis, Lactobacillus acidophilus, Bacteroides ureolyticus and Staphylococcus epidermidis (1 x 10° cells/assay) were added to samples containing the rRNA equivalent of approximately 1 C. trachomatis IFU (5 fg) and 50 N. gonorrhoeae cells (250 fg). These additions did not interfere with the amplification and detection of C. trachomatis or N. gonorrhoeae rRNA using the AC2 Assay.

# PreservCyt Liquid Pap Specimen Stability Studies

Data to support the recommended shipping and storage conditions for PreservCyt liquid Pap samples were generated with pooled negative PreservCyt liquid Pap samples. Four pooled samples were spiked with C. trachomatis and N. gonorrhoeae at final concentrations of 10 IFU and 100 CFU per reaction, respectively. The PreservCyt liquid Pap samples were placed at 30℃ for 7 days, after which 1 mL of the sample was added to an APTIMA Transfer Tube. The spiked samples were held at 4℃, 10℃ and 30℃. Samples stored at 4℃ and 10℃ were tested in duplicate at days 0, 6, 13, 26, 30 and 36. Samples stored at 30℃ were tested in duplicate at days 0, 5, 8, 14 and 17. Four spiked PreservCyt Iiquid Pap sample pools were added to APTIMA Transfer Tubes and placed at 30℃ for 14 days before being stored at -20℃. The -20℃ samples were tested in duplicate after 0, 30, 60, 90 and 106 days of storage. All test conditions were positive for both C. trachomatis and N. gonorrhoeae at all times and temperatures.

#### Precision

Reproducibility was established with a 12-member panel generated by spiking PreservCyt Solution with 0 to 2000 fg/assay of C. trachomatis and 0 to 5,000 fg/assay of N. gonorrhoeae rRNA and aliquotting 1mL into the APTIMA Specimen Transfer Kit collection tube. Two (2) operators at each of the three sites performed one run per day on each of three days, totaling three valid runs per operator. The results of the precision study are summarized in the table below. Reproducibility when testing PreservCyt liquid Pap clinical specimens containing target organism has not been determined.

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| | | Inter-Site | | Inter-Operator | | Inter-Run | | Intra-Run | |
|----------|---------------------|---------------|-------|----------------|-------|---------------|-------|---------------|-------|
| Agrmt(%) | Mean RLU<br>(x1000) | SD<br>(x1000) | CV(%) | SD<br>(x1000) | CV(%) | SD<br>(x1000) | CV(%) | SD<br>(x1000) | CV(%) |
| 97.5 | 9.7 | 6.4 | N/A | 4.7 | N/A | 3.4 | N/A | 31.6 | N/A |
| 96.3 | 1296 | 0.0 | 0.0 | 0.0 | 0.0 | 54.8 | 4.2 | 146 | 11.3 |
| 100 | 1140 | 101 | 8.9 | 2.4 | 0.2 | 79.8 | 7.0 | 54.1 | 4.7 |
| 100 | 2345 | 94.7 | 4.0 | 37.9 | 1.6 | 78.0 | 3.3 | 79.6 | 3.4 |
| 100 | 953 | 161 | 16.9 | 90.7 | 9.5 | 0.0 | 0.0 | 114 | 12.0 |
| 100 | 971 | 22.8 | 2.4 | 85.0 | 8.8 | 71.7 | 7.4 | 58.3 | 6.0 |
| 100 | 2294 | 153 | 6.7 | 0.0 | 0.0 | 88.9 | 3.9 | 114 | 5.0 |
| 98.1 | 1911 | 348 | 18.2 | 39.7 | 2.1 | 130 | 6.8 | 139 | 7.3 |
| 100 | 2136 | 98.8 | 4.6 | 166 | 7.8 | 130 | 6.1 | 113 | 5.3 |
| 96.3 | 2044 | 360 | 17.6 | 26.9 | 1.3 | 169 | 8.3 | 138 | 6.7 |

TIMA Combo 2 Assay Precision Study - PreservCyt Solutio

W/A = not applicable for negative panel members
Agnnt = Agreement, CV = Coefficient of variation, RLU = Relative Light Units, SD = Standard deviation

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### PreservCyt Liquid Pap Specimen Clinical Study Results

A prospective multi-center clinical study was conducted to evaluate the use of the PreservCyt Solution (a component of the ThinPrep 2000 System) as an alternative medium for gynecological specimens for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae. One thousand six hundred forty-seven (1,647) symptomatic and asymptomatic female subjects attending OB/GYN, family planning, public health, women's and STD clinics were evaluated in the clinical study. Of the 1,647 evaluable subjects, 1,288 were asymptomatic subjects and 359 were symptomatic subjects. Subjects were enrolled from sites with CT prevalence that ranged from 3.2 to 14.0% and GC prevalence that ranged from 0 to 5.0%. Two specimens were collected from each eligible subject: one PreservCyt Solution specimen and one endocervical swab. PreservCyt Solution specimens were processed in accordance with the ThinPrep 2000 Processor Operator's Manual and APTIMA Specimen Transfer Kit Package Insert. After processing the PreservCyt Solution specimen with the ThinPrep 2000 Processor, the specimen was transferred into the APTIMA Specimen Transfer Kit for testing with the APTIMA Combo 2 Assay. The PreservCyt liquid Pap specimens and endocervical swab specimens were tested with the APTIMA Combo 2 Assay.

Sensitivity and specificity for PreservCyt liquid Pap specimens were calculated by comparing results to a patient infected status algorithm. In the algorithm, the designation of a subject as being infected or non-infected with C. trachomatis or N.gonorrhoeae was based on endocervical swab specimen results from two commercially-available NAATs. For C. trachomatis, the reference NAATs included the APTIMA Combo 2 Assay and the APTIMA CT Assay. For N. gonorrhoeae, the reference NAATs included the APTIMA Combo 2 Assay and the APTIMA GC Assay. Positive results from both reference NAATs were required to establish an infected patient. A non-infected patient was established if the results from the two reference NAATs disagreed or were negative.

Sensitivity and specificity for C. trachomatis in PreservCyt liquid Pap specimens tested in the APTIMA Combo 2 Assay, by symptom status and overall, is presented in Table 1. For

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C. trachomatis, overall sensitivity was 96.7% (87/90). In symptomatic and asymptomatic subjects, sensitivity was 96.7% (29/30) and 96.7% (58/60), respectively. Overall specificity for C. trachomatis PreservCyt liquid Pap specimens was 99.2% (1545/1557). In symptomatic and asymptomatic subjects, specificity was 98.5% (324/329) and 99.4% (1221/1228), respectively. Table 2 shows the APTIMA Combo 2 Assay sensitivity and specificity values for C. trachomatis in PreservCyt liquid Pap specimens by clinical site and overall. For C. trachomatis, the sensitivity ranged from 92.9% to 100%. The specificity ranged from 97.7% to 100%.

Sensitivity and specificity for N. gonorrhoeae in PreservCyt liquid Pap specimens tested in the APTIMA Combo 2 Assay, by symptom status and overall, is presented in Table 3. For N. gonorrhoeae, overall sensitivity was 92.3% (12/13). In symptomatic and asymptomatic subjects, sensitivity was 100% (7/7) and 83.3% (5/6), respectively. Overall specificity for N. gonorrhoeae PreservCyt liquid Pap specimens was 99.8% (1630/1633). symptomatic and asymptomatic subjects, specificity was 100% (352/352) and 99.8% (1278/1281), respectively. Table 4 shows the APTIMA Combo 2 Assay sensitivity and specificity values for N. gonorrhoeae in PreservCyt liquid Pap specimens by clinical site and overall. For N. gonorrhoeae, the sensitivity ranged from 80.0% to 100%. Specificity ranged from 99.0% to 100%.

The distribution of cervical sampling devices used in this clinical study according to clinical site is summarized in the table below.

## Cervical sampling devices

| Cervical sampling device used | Clinical Collection Site | | | | | | Total |
|-------------------------------|--------------------------|-----|-----|-----|-----|-----|-------|
| | 1 | 2 | 3 | 4 | 5 | 6 | |
| Spatula/Cytobrush | 0 | 124 | 475 | 287 | 57 | 364 | 1307 |
| Broom-Type Device | 100 | 0 | 0 | 0 | 240 | 0 | 340 |

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| | | AC2 ES+ and ACT ES+ | | | AC2 ES+ and ACT ES- | | | Sensitivity (95% CI) | | Specificity (95% CI) | |
|---------------------------------------------------------------------------|---------------------------|---------------------|----------|-------|---------------------|----------|-------|----------------------|-------------------|----------------------|--|
| | AC2/CT<br>ThinPrep Result | Positive | Negative | Total | Positive | Negative | Total | | | | |
| Asympt | Positive | 58 | 1 | 60 | 0 | 1 | 2 | 96.7% (88.5-99.6) | 99.4% (98.8-99.8) | | |
| | Negative | 2 | 1 | 3 | 6 | 12 | 1208 | | | | |
| | Total | 60 | 2 | 12 | 1214 | | | | | | |
| Sympt | Positive | 29 | 0 | 29 | 0 | 3 | 3 | 96.7% (82.8-99.9) | 98.5% (96.5-99.5) | | |
| | Negative | 1 | 3 | 4 | 5 | 4 | 317 | | | | |
| | Total | 30 | 3 | 332 | 4 | 322 | | | | | |
| All | Positive | 87 | 1 | 88 | 0 | 1 | 1 | 96.7% (90.6-99.3) | 99.2% (98.7-99.6) | | |
| | Negative | 3 | 4 | 7 | 11 | 16 | 1525 | | | | |
| | Total | 90 | 5 | 16 | 1536 | | | | | | |
| AC2 ES+ = Positive endocervical swab specimen APTIMA Combo 2 Assay result | | | | | | | | | | | |

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ACE BS - - Positive endocervical swab specimen APTIMA Cosmo 2 rossil
ACT ES+ = Negative endocervical swab specimen APTIMA CT Assalt
ACT ES+ = Positive endocervical swab speci

Endocervical Swab Specimen, Negative == RESULTS

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| Site | AC2/CT<br>ThinPrep Result | AC2 ES+<br>and<br>ACT ES+ | AC2 ES+<br>and<br>ACT ES- | AC2 ES-<br>and<br>ACT ES+ | AC2 ES-<br>and<br>ACT ES- | Prev<br>(%) | Sensitivity (95% CI) (%) | Specificity (95% CI) (%) | PPV<br>(%) | NPV<br>(%) |
|------|---------------------------|---------------------------|---------------------------|---------------------------|---------------------------|-------------|--------------------------|--------------------------|------------|------------|
| 1 | Positive | 14 | 0 | 0 | 2 | 14.0 | 100% (76.8 - 100) | 97.7% (91.9 - 99.7) | 87.5 | 100 |
| | Negative | 0 | 0 | 1 | 83 | | | | | |
| | Total | 14 | 0 | 1 | 85 | | | | | |
| 2 | Positive | 4 | 0 | 0 | 0 | 3.2 | 100% (39.8 - 100) | 100% (97.0 - 100) | 100 | 100 |
| | Negative | 0 | 0 | 2 | 118 | | | | | |
| | Total | 4 | 0 | 2 | 118 | | | | | |
| 3 | Positive | 29 | 0 | 0 | 2 | 6.5 | 93.5% (78.6 - 99.2) | 99.5% (98.4 - 99.9) | 93.5 | 99.5 |
| | Negative | 2 | 0 | 2 | 440 | | | | | |
| | Total | 31 | 0 | 2 | 442 | | | | | |
| 4 | Positive | 8 | 1 | 0 | 4 | 2.8 | 100% (63.1 - 100) | 98.2% (95.9 - 99.4) | 61.5 | 100 |
| | Negative | 0 | 2 | 1 | 271 | | | | | |
| | Total | 8 | 3 | 1 | 275 | | | | | |
| 5 | Positive | 13 | 0 | 0 | 2 | 4.7 | 92.9% (66.1 - 99.8) | 99.3% (97.5 - 99.9) | 86.7 | 99.6 |
| | Negative | 1 | 1 | 4 | 276 | | | | | |
| | Total | 14 | 1 | 4 | 278 | | | | | |
| 6 | Positive | 19 | 0 | 0 | 1 | 5.2 | 100% (82.4 - 100) | 99.7% (98.4 - 100) | 95.0 | 100 |
| | Negative | 0 | 1 | 6 | 337 | | | | | |
| | Total | 19 | 1 | 6 | 338 | | | | | |
| All | Positive | 87 | 1 | 0 | 11 | 5.5 | 96.7% (90.6 - 99.3) | 99.2% (98.7 - 99.6) | 87.9 | 99.8 |
| | Negative | 3 | 4 | 16 | 1525 | | | | | |
| | Total | 90 | 5 | 16 | 1536 | | | | | |

ACE ES - Tostive endocervical swab specimen APTIMA Combro 2 Assay result
ACT ES+ = Negative endocervical swab specimen AFTIMA CT Assay result
ACT ES+ = Positive endocervical

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| Table 3: GC Sensitivity and Specificity: AC2 Assay PreservCyt Specimens vs. Patient Infected Status | AC2/GC<br>ThinPrep Result | AC2 ES+<br>and<br>AGC ES+ | AC2 ES-<br>and<br>AGC ES+ | AC2 ES+<br>and<br>AGC ES- | AC2 ES-<br>and<br>AGC ES- | Sensitivity (95% CI) | Specificity (95% CI) |
|-----------------------------------------------------------------------------------------------------|---------------------------|---------------------------|---------------------------|---------------------------|---------------------------|----------------------|----------------------|
| Asympt | Positive | 5 | 0 | 0 | 3 | 83.3% (35.9 - 99.6) | 99.8% (99.3 - 100) |
| | Negative | 1 | 0 | 5 | 1273 | | |
| | Total | 6 | 0 | 5 | 1276 | | |
| Sympt | Positive | 7 | 0 | 0 | 0 | 100% (59.0 - 100) | 100% (99.0-100) |
| | Negative | 0 | 0 | 0 | 352 | | |
| | Total | 7 | 0 | 0 | 352 | | |
| All | Positive | 12 | 0 | 0 | 3 | 92.3% (64.0 - 99.8) | 99.8% (99.5 - 100) |
| | Negative | 1 | 0 | 5 | 1625 | | |
| | Total | 13 | 0 | 5 | 1628 | | |

2 ES+= Positive endocervical swab specimen Al
C2
MA Combo 2 Assay resu
ES-=Negative endocervical swab specimen APTIMA Combo 2 Assay resu

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| Site | AC2/GC<br>ThinPrep Result | AC2 ES+<br>and<br>AGC ES+ | AC2 ES+<br>and<br>AGC ES- | AC2 ES-<br>and<br>AGC ES+ | AC2 ES-<br>and<br>AGC ES- | Prev<br>(%) | Sensitivity (95% CI) | Specificity (95% CI) | PPV<br>(%) | NPV<br>(%) |
|------|---------------------------|---------------------------|---------------------------|---------------------------|---------------------------|-------------|----------------------|----------------------|------------|------------|
| 1 | Positive | 5 | 0 | 0 | 0 | 5.0 | 100% (47.8 - 100) | 100% (96.2 - 100) | 100 | 100 |
| | Negative | 0 | 0 | 0 | 95 | | | | | |
| | Total | 5 | 0 | 0 | 95 | | | | | |
| 2 | Positive | 1 | 0 | 0 | 0 | 0.8 | 100% (2.5 - 100) | 100% (97.0 - 100) | 100 | 100 |
| | Negative | 0 | 0 | 0 | 123 | | | | | |
| | Total | 1 | 0 | 0 | 123 | | | | | |
| 3 | Positive | 4 | 0 | 0 | 0 | 1.1 | 80.0% (28.4 - 99.5) | 100% (99.2 - 100) | 100 | 99.8 |
| | Negative | 1 | 0 | 0 | 470 | | | | | |
| | Total | 5 | 0 | 0 | 470 | | | | | |
| 4 | Positive | 1 | 0 | 0 | 0 | 0.3 | 100% (2.5 - 100) | 100% (98.7 - 100) | 100 | 100 |
| | Negative | 0 | 0 | 3 | 283 | | | | | |
| | Total | 1 | 0 | 3 | 283 | | | | | |
| 5 | Positive | 0 | 0 | 0 | 3 | 0.0 | N/A | 99.0% (97.1 - 99.8) | 0.0 | 100 |
| | Negative | 0 | 0 | 0 | 294 | | | | | |
| | Total | 0 | 0 | 0 | 297 | | | | | |
| 6 | Positive | 1 | 0 | 0 | 0 | 0.3 | 100% (2.5 - 100) | 100% (99.0 - 100) | 100 | 100 |
| | Negative | 0 | 0 | 2 | 360 | | | | | |
| | Total | 1 | 0 | 2 | 360 | | | | | |
| All | Positive | 12 | 0 | 0 | 3 | 0.8 | 92.3% (64.0 - 99.8) | 99.8% (99.5 - 100) | 80.0 | 99.9 |
| | Negative | 1 | 0 | 5 | 1625 | | | | | |
| | Total | 13 | 0 | 5 | 1628 | | | | | |

| Total | 13 | 0 | 5 | 1028 |
|--------------------------------------------------------------------------|----|---|---|------|
| AC2 ES+= Positive endocervical swab specimen APTIMA Combo 2 Assay result | | | | |
| AC2 ES-= Negative endocervical swab specimen APTIMA Combo 2 Assay result | | | | |
| AGC ES+= Positive endocervical swab specimen APTIMA GC Assay result | | | | |
| AGC ES-= Negative endocervical swab specimen APTIMA GC Assay result | | | | |
| N/A = not applicable | | | | |

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### Prevalence

The prevalence of C. trachomatis and/or N. gonorrhoeae disease in patient populations depends on risk factors such as age, gender, the presence of symptoms, the type of clinic, and the test method. A summary of the prevalence of three C. trachomatis and N. gonorrhoeae disease outcomes as determined by the APTIMA Combo 2 Assay is shown in Table 5 by clinical site and overall. The CT and GC prevalence were calculated using the APTIMA Combo 2 Assay results of PreservCyt liquid Pap specimen

Table 5: Prevalence of C. trachomatis and/or N. gonorrhoeae Disease as Determined by the APTIMA Combo 2 Assay Results by Clinical Site

| Site | PreservCyt liquid Pap Specimens<br>% Prevalence (# positive/# tested) | | |
|------|-----------------------------------------------------------------------|------------------|------------------|
| | CT+/GC+ | CT+/GC- | CT-/GC+ |
| 1 | 3.0<br>(3/100) | 13.0<br>(13/100) | 2.0<br>(2/100) |
| 2 | 0<br>(0/124) | 3.2<br>(4/124) | 0.8<br>(1/124) |
| 3 | 0.4<br>(2/475) | 6.1<br>(29/475) | 0.4<br>(2/475) |
| 4 | 0.4<br>(1/287) | 4.2<br>(12/287) | 0<br>(0/287) |
| 5 | 0<br>(0/297) | 5.1<br>(15/297) | 1.0<br>(3/297) |
| 6 | 0<br>(0/364) | 5.5<br>(20/364) | 0.6<br>(2/364) |
| All | 0.4<br>(6/1647) | 5.6<br>(93/1647) | 0.6<br>(10/1647) |

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

## Conclusions from the Clinical Data

The non clinical and clinical study results support the use of PreservCyt liquid Pap specimens collected and processed by the Cytyc ThinPrep 2000 Processor in the GEN-PROBE APTIMA Combo 2 Assay for the detection of C. trachomatis and/or N. gonorrhoeae. The GEN-PROBE APTIMA Specimen Transfer Kit provides the necessary materials and instructions to allow for the testing of PreservCyt liquid Pap specimens in the AC2 Assay. Use of this ancillary kit broadens the application of the AC2 Assay as a diagnostic tool to provide information that measurably contributes to a diagnosis of C. trachomatis and/or N. gonorrhoeae infection.

The results of the clinical study demonstrate reasonable evidence that when the AC2 Assay and the APTIMA Specimen Transfer Kit are labeled as proposed, the AC2 Assay continues to be safe and effective for its stated intended use.

## Contraindications and Cautions

There are no contraindications or cautions.

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

Image /page/16/Picture/1 description: The image shows a logo with a circular border containing the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA". Inside the circle is a stylized graphic of an eagle or bird-like figure, represented by three curved lines that suggest wings or feathers. The logo is simple and monochromatic, with a focus on the text and the abstract bird symbol.

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

AUG 9 - 2005

Alan Maderazo, Ph.D. RAC Sr. Regulatory Affairs Specialist Gen-Probe Incorporated 10210 Genetic Center Drive San Diego, CA 92121-1589

Re: K043224

Trade/Device Name: GEN-PROBE® APTIMA® Combo 2 Assay Regulation Number: 21 CFR 866.3390 Regulation Name: Neisseria spp. direct serological test reagents Regulatory Class: Class II Product Code: LSL, MKZ Dated: August 2, 2005 Received: August 4, 2005

Dear Dr. Maderazo:

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), it may be subject to such additional controls. Existing major regulations affecting your device can be found in Title 21, Code of Federal Regulations (CFR), Parts 800 to 895. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); and good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820).

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

Page 2 -

This letter will allow you to begin marketing your device as described in your Section 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.

If you desire specific information about the application of labeling requirements to your device, or questions on the promotion and advertising of your device, please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at [phone]. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). 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,

Sales, a Form

Sally A. Hojvat, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

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

## CDRH Special Sheet - Device Indications for Use

510(k) Number (if known): K043224

Device Name: GEN-PROBE® APTIMA® Combo 2 Assay

Indications For Use:

#### AC2 Assay package insert:

The APTIMA Combo 2 Assay is a target amplification nucleic acid probe test that utilizes target capture for the in vitro qualitative detection and differentiation of ribosomal RNA (rRNA) from Chlamydia trachomatis (CT) and/or Neisseria gonorrhoeae (GC) in clinician-collected endocervical, and male urethral swab specimens, patient-collected vaginal swab specimens , and female and male urine specimens. The assay is also intended for use with testing of gynecological specimens collected in the PreservCyt Solution and processed with the Cytyc ThinPrep 2000 System. The assay may be used to test specimens from symptomatic and asymptomatic individuals to aid in the diagnosis of gonococcal and/or chlamydial urogenital disease.

*Patient-collected vaginal swab specimens are an option for screening women when a pelvic exam is not otherwise indicated. The vaginal swab specimen collection kit is not for home use.

#### Ancillary Kit package insert:

The GEN-PROBE® APTIMA® Specimen Transfer Kit is only for use with GEN-PROBE APTIMA assays for the detection of Chlamydia trachomatis and/or Neisseria gonorrhoeae. The GEN-PROBE APTIMA Specimen Transfer Kit allows for APTIMA Assay testing of gynecological specimens collected and processed by the Cytyc ThinPrep 2000 Processor according to the instructions provided.

Prescription Use _____________________________________________________________________________________________________________________________________________________________ (Part 21 CFR 801 Subpart D)

AND/OR

Over-The-Counter Use _ (21 CFR 807 Subpart C)

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

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

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Office of In Vitro Diagnostic Device Evaluation and Safety

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| 510(k) | k043224 |
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| Confidential | |