Predicate Devices

Reference Devices

AI/ML (Artificial Intelligence / Machine Learning)

No
The device description focuses on standard molecular diagnostic techniques (target capture, TMA, DKA, chemiluminescent detection) and does not mention any AI or ML components for data analysis or interpretation. The results are determined by a cut-off based on RLU and kinetic curve type, which is a rule-based approach, not ML.

Therapeutic

No.
Explanation: This device is for the in vitro qualitative detection and differentiation of ribosomal RNA to aid in the diagnosis of chlamydial and/or gonococcal urogenital disease. It is a diagnostic device, not a therapeutic one.

Diagnostic

Yes

Explanation: The "Intended Use / Indications for Use" section explicitly states that the device is used "to aid in the diagnosis of chlamydial and/or gonococcal urogenital disease." This directly defines its purpose as diagnostic.

SaMD (Software as a Medical Device)

No

The device description clearly outlines a laboratory-based assay involving physical specimens, reagents, magnetic microparticles, and a PANTHER System (hardware) for target capture, amplification, and detection using chemiluminescence. This is not a software-only device.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

Here's why:

Intended Use: The "Intended Use / Indications for Use" section explicitly states that the APTIMA COMBO 2 Assay is for "in vitro qualitative detection and differentiation of ribosomal RNA (rRNA) from Chlamydia trachomatis (CT) and/or Neisseria gonorrhoeae (GC) to aid in the diagnosis of chlamydial and/or gonococcal urogenital disease". The phrase "in vitro" is a key indicator of an IVD, meaning it is used to test samples outside of the body.
Device Description: The description details how the assay processes biological specimens (swabs, PreservCyt solution) in a laboratory setting using various reagents and technologies (target capture, TMA, DKA) to detect specific nucleic acids. This process is characteristic of an in vitro diagnostic test.
Specimen Types: The assay is designed to test various biological specimens collected from individuals (endocervical, vaginal, male urethral swabs, PreservCyt solution).
Purpose: The purpose is to "aid in the diagnosis" of specific diseases (chlamydial and/or gonococcal urogenital disease), which is a diagnostic function performed on samples outside the body.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for 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) to aid in the diagnosis of chlamydial and/or gonococcal urogenital disease using the PANTHER System as specified.

On the PANTHER System, the assay may be used to test the following specimens from symptomatic and asymptomatic individuals: clinician-collected endocervical, vaginal and male urethral swab specimens, clinician-collected gynecological specimens collected in the PreservCyt Solution, and patient-collected vaginal swab specimens.

'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.

Product codes (comma separated list FDA assigned to the subject device)

LSL, MKZ, NSU

Device Description

The APTIMA Combo 2 Assay combines the technologies of target capture, TMA, and DKA.

Specimens are collected and transferred into their respective specimen transport tubes. The transport solutions in these tubes release the rRNA targets and protect them from degradation during storage. When the APTIMA Combo 2 Assay is performed in the laboratory, the farget rRNA molecules are isolated from specimens by use of capture oligomers via target capture that utilizes magnetic microparticles. The capture oligomers contain sequences complementary to specific regions of the target molecules as well as a string of deoxyadenosine residues. A separate capture oligomer is used for each target. During the hybridization step, the sequence specific regions of the capture oligomers bind to specific regions of the target molecules. The capture oligomer:target complex is then captured out of solution by decreasing the temperature of the reaction to room temperature. This temperature reduction allows hybridization to occur between the deoxyadenosine region on the capture oligomer and the poly-deoxythymidine molecules that are covalently attached to the magnetic particles. The microparticles, including the captured target molecules bound to them, are pulled to the side of the reaction vessel using magnets and the supernatant is aspirated. The particles are washed to remove residual specimen matrix that may contain amplification reaction inhibitors. After the target capture steps are completed, the specimens are ready for amplification.

Target amplification assays are based on the ability of complementary oligonucleotide primers to specifically anneal and allow enzymatic amplification of the target nucleic acid strands. The APTIMA Combo 2 Assay replicates a specific region of the 23S rRNA from CT and a specific region of the 16S rRNA from GC via DNA intermediates. A unique set of primers is used for each target molecule. Detection of the rRNA amplification product sequences (amplicon) is achieved using nucleic acid hybridization. Single-stranded chemiluminescent DNA probes, which are complementary to a region of each target amplicon, are labeled with different acridinium ester molecules. The labeled DNA probes combine with amplicon to form stable RNA:DNA hybrids. The Selection Reagent differentiates hybridized from unhybridized probe, eliminating the generation of signal from unhybridized probe. 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). In DKA, differences in the kinetic profiles of the CT and GC labeled probes allow for the differentiation of signal; kinetic profiles are derived from measurements of photon output during the detection read time. The chemiluminescent detection for CT signal has very rapid kinetics and has the "flasher" kinetic type. The chemiluminescent detection reaction for GC signal is relatively slower and has the "glower" kinetic type. Assay results are determined by a cut-off based on the total RLU and the kinetic curve type.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Urogenital

Indicated Patient Age Range

Not Found

Intended User / Care Setting

Not Found

Description of the training set, sample size, data source, and annotation protocol

Not Found

Description of the test set, sample size, data source, and annotation protocol

A prospective, multicenter clinical study was conducted to establish the performance characteristics of the APTIMA COMBO 2 Assay on the PANTHER System. Specimens were collected from symptomatic and asymptomatic men (n=580) and women (n=1332) enrolled from 7 geographically and ethnically diverse US clinical sites, including obstetrics and gynecology, family planning, public health, and STD clinics. Subjects were classified as symptomatic if symptoms were reported by the subject. Subjects were classified as asymptomatic if the subject did not report symptoms.

Of the 580 male subjects, none were 25 years of age. Of the 1332 female subjects, 11 were 14 to 15 years of age, 59 were 16 to 17 years of age, 319 were 18 to 20 years of age, 401 were 21 to 25 years of age, and 542 were >25 years of age.

Up to 2 specimens were collected from each male subject (1 urethral swab and 1 first-catch urine, in that order) and up to 4 specimens were collected from each female subject (1 first-catch urine, 1 vaginal swab, 1 PreservCyt Solution liquid Pap specimen, and 1 endocervical swab, in that order). All specimens were clinician-collected except urine specimens and approximately half of the vaginal swab specimens, which were collected by the subject at the clinic. Approximately half of the PreservCyt Solution liquid Pap specimens were collected with a broom-type device and half were collected with a spatula and cytobrush. Samples were prepared for APTIMA testing in accordance with the appropriate APTIMA specimen collection kit package insert instructions.

All evaluable samples (567 male urethral swab, 1319 vaginal swab, 1330 PreservCyt, and 1310 endocervical swab samples) were tested with the APTIMA COMBO 2 Assay on the PANTHER System in accordance with package insert instructions. The samples were split amongst three laboratories (two external laboratories and GEN-PROBE). Samples with initial invalid, equivocal, or error results were retested. Eighteen (18) male urethral swab, 25 vaginal swab, 1 PreservCyt, and 37 endocervical swab specimens had final invalid results and were excluded from the analyses. Most of the invalid results were due to insufficient sample volume. One vaginal swab and 1 endocervical swab had final CT equivocal results and 1 PreservCyt and 1 endocervical swab had final GC equivocal results and were excluded from the analyses.

Male urethral swab, male and female urine, and PreservCyt samples were tested with cleared nucleic acid amplification tests (NAATs) to establish the infected status. The infected status algorithm used results from two specimen types and two reference NAATs. Subjects were categorized as infected if a positive result occurred in each of the 2 reference NAATs. For female subjects, if the positive NAAT results occurred only in the urine specimens and not in the PreservCyt specimens, the subject was categorized as infected; however, for the evaluation of the non-urine specimen types, the specimens were considered non-infected. Subjects that could not be categorized as infected or not infected were excluded from the performance analyses.

Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)

Analytical Sensitivity Study

Chlamydia trachomatis analytical sensitivity (limits of detection) was determined by directly comparing dilutions of CT organisms in the APTIMA COMBO 2 assay. The analytical sensitivity claim for the assay is 1 IFU/assay (7.25 IFU/swab, 9.75 IFU/mL, PreservCyt Solution liquid Pap). However, dilutions of less than 1 IFU/assay tested positive in the APTIMA COMBO 2 Assay (for this study, 100% positivity was observed in samples containing CT concentrations of 0.03 IFU/mL).
Neisseria gonorrhoeae analytical sensitivity (limits of detection) was determined by directly comparing dilutions of GC organisms in the APTIMA COMBO 2 Assay. The analytical sensitivity claim for the assay is 50 cells/assay (362 cells/swab, 488 cells/mL PreservCyt Solution liquid Pap). However, dilutions of less than 50 cells/assay tested positive in the APTIMA COMBO 2 Assay (for this study, 100% positivity was observed in samples containing GC concentrations of 0.04 CFU/mL).

Within Laboratory Precision Study

Sample size: Not explicitly stated for each concentration, but testing was performed over 24 days with three PANTHER Systems and three lots of assay reagents. Reproducibility panel members were created using negative PreservCyt Solution liquid Pap specimens and STM, spiked with CT and/or GC organisms.
Results for CT: Mean RLU values range from 6 to 2496 (x1000). Total CV% ranges from 3.4% to 20.1%. Percent agreement with expected results was 100% across all CT concentrations tested.
Results for GC: Mean RLU values range from 7 to 2471 (x1000). Total CV% ranges from 3.3% to 24.7%. Percent agreement with expected results was 100% across all GC concentrations tested.

Carryover Studies for the PANTHER System

A multi-run analytical study was conducted using spiked panels on three PANTHER Systems.
Sample size: Total of 2938 valid negative results.
Results: The overall carryover rate was 0% with a 95% confidence interval of 0-0.1%.

Clinical Study Results

Study type: Prospective, multicenter clinical study.
Sample size: 549 Male Urethral Swab, 1274 Clinician-Collected Vaginal Swab/Patient-Collected Vaginal Swab, 1311 PreservCyt Solution liquid Pap, 1254 Female Endocervical Swab for CT detection. Similar sample sizes for GC detection. Individual sites and symptom statuses also reported, with varying sample numbers.
Key results for CT detection:
- Male Urethral Swab: Sensitivity 100%, Specificity 99.1%, PPV 96.2%, NPV 100%.
- CVS/PVS: Sensitivity 97.2%, Specificity 98.5%, PPV 85.2%, NPV 99.7%.
- PreservCyt: Sensitivity 98.2%, Specificity 100%, PPV 100%, NPV 99.8%.
- FS: Sensitivity 97.2%, Specificity 99.3%, PPV 92.9%, NPV 99.7%.
Key results for GC detection:
- Male Urethral Swab: Sensitivity 100%, Specificity 100%, PPV 100%, NPV 100%.
- VS: Sensitivity 97.7%, Specificity 99.6%, PPV 89.4%, NPV 99.9%.
- PreservCyt: Sensitivity 100%, Specificity 100%, PPV 100%, NPV 100%.
- FS: Sensitivity 100%, Specificity 99.8%, PPV 95.5%, NPV 100%.

Reproducibility Study

Study type: Multi-site reproducibility study (two external US laboratories and Gen-Probe).
Sample size: Testing was performed over at least 10 days at each site using one lot of assay reagents and a total of six operators. 180 samples in most categories for each target concentration.
Results for CT: Mean RLU values range from 6.2 to 1278.5 (x1000). Total CV% ranges from 9.4% to 133.0%. Percent agreement with expected results was 98.9% to 100%.
Results for GC: Mean RLU values range from 421.9 to 2503.8 (x1000). Total CV% ranges from 3.2% to 23.0%. Percent agreement with expected results was 98.9% to 100%.

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

CT Detection:

Male Urethral Swab:
- Sensitivity: 100% (95% CI: 96.3-100)
- Specificity: 99.1% (95% CI: 97.7-99.7)
- PPV: 96.2% (95% CI: 90.8-98.9)
- NPV: 100% (95% CI: 99.2-100)
Clinician-Collected/Patient-Collected Vaginal Swab (CVS/PVS):
- Sensitivity: 97.2% (95% CI: 92.1-99.0)
- Specificity: 98.5% (95% CI: 97.6-99.0)
- PPV: 85.2% (95% CI: 78.8-90.5)
- NPV: 99.7% (95% CI: 99.3-99.9)
PreservCyt Solution Liquid Pap (PCyt):
- Sensitivity: 98.2% (95% CI: 93.8-99.5)
- Specificity: 100% (95% CI: 99.7-100)
- PPV: 100% (95% CI: 96.9-100)
- NPV: 99.8% (95% CI: 99.4-100)
Female Endocervical Swab (FS):
- Sensitivity: 97.2% (95% CI: 92.1-99.0)
- Specificity: 99.3% (95% CI: 98.6-99.6)
- PPV: 92.9% (95% CI: 87.1-96.7)
- NPV: 99.7% (95% CI: 99.3-99.9)

GC Detection:

Male Urethral Swab (MS):
- Sensitivity: 100% (95% CI: 89.8-100)
- Specificity: 100% (95% CI: 99.3-100)
- PPV: 100% (95% CI: 90.2-100)
- NPV: 100% (95% CI: 99.3-100)
Vaginal Swab (VS) (Likely clinician-collected/patient-collected as in CT section):
- Sensitivity: 97.7% (95% CI: 87.9-99.6)
- Specificity: 99.6% (95% CI: 99.0-99.8)
- PPV: 89.4% (95% CI: 78.6-96.1)
- NPV: 99.9% (95% CI: 99.6-100)
PreservCyt Solution Liquid Pap (PCyt):
- Sensitivity: 100% (95% CI: 91.8-100)
- Specificity: 100% (95% CI: 99.7-100)
- PPV: 100% (95% CI: 92.1-100)
- NPV: 100% (95% CI: 99.7-100)
Female Endocervical Swab (FS):
- Sensitivity: 100% (95% CI: 91.6-100)
- Specificity: 99.8% (95% CI: 99.4-100)
- PPV: 95.5% (95% CI: 85.4-99.4)
- NPV: 100% (95% CI: 99.7-100)

Reproducibility (Overall %CV and % Agreement with Expected Results):

CT:
- Negative (0 IFU/mL): Overall CV 133.0%, Agreement 98.9%
- 0.25 IFU/mL: Overall CV 10.2%, Agreement 100%
- 2.5 IFU/mL: Overall CV 9.4%, Agreement 100%
- 25 IFU/mL: Overall CV 9.7%, Agreement 100%
- 1000 IFU/mL: Overall CV 9.8%, Agreement 100%
GC:
- 0.25 CFU/mL: Overall CV 23.0%, Agreement 98.9%
- 12.5 CFU/mL: Overall CV 8.4%, Agreement 99.4%
- 125 CFU/mL: Overall CV 4.4%, Agreement 100%
- 1250 CFU/mL: Overall CV 3.5%, Agreement 100%
- 2500 CFU/mL: Overall CV 3.2%, Agreement 100%
CT and GC Combined:
- CT 2.5/GC 125: Overall CV 4.6%, Agreement 100%
- CT 2.5/GC 2500: Overall CV 4.7%, Agreement 100%
- CT 1000/GC 125: Overall CV 4.4%, Agreement 100%
- CT 1000/GC 2500: Overall CV 5.2%, Agreement 100%

Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.

APTIMA Combo 2 Assay; K032194

Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.

K060652, K061413, K061509

Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).

Not Found

Regulation Number and Section

§ 866.3390

Neisseria spp. direct serological test reagents.(a)
Identification. Neisseria spp. direct serological test reagents are devices that consist of antigens and antisera used in serological tests to identifyNeisseria spp. from cultured isolates. Additionally, some of these reagents consist ofNeisseria spp. antisera conjugated with a fluorescent dye (immunofluorescent reagents) which may be used to detect the presence ofNeisseria spp. directly from clinical specimens. The identification aids in the diagnosis of disease caused by bacteria belonging to the genusNeisseria, 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.(b)
Classification. Class II (performance standards).

Summary

0

K111409

Gen-Probe Incorporated

510(k) SUMMARY

MAY - 3 2012

APTIMA Combo 2® Assay on the PANTHER System

Sponsor/Contact Information

Submitted By:

Name: Address: Gen-Probe Incorporated 10210 Genetic Center Drive San Diego, CA 92121 (858) 410-8000

Company Contact:

Jody J. Fleming Contact: Regulatory Affairs Manager Phone: 858-410-8634 Fax: 858-410-7876 Email: jody.fleming@gen-probe.com

Date Prepared:

May 1, 2012

1

General Information

Trade Name:	APTIMA Combo 2® Assay
Common or Usual Name:	Ribosomal RNA (rRNA) target-amplified nucleic acid probe
test for the in vitro diagnostic detection of Chlamydia
trachomatis and/or Neisseria gonorrhoeae
Classification Names:	DNA Probe, Nucleic Acid Amplification, Chlamydia
DNA Reagents, Neisseria

APTIMA Combo 2 Assay

Device Description	DNA Probe, Nucleic Acid Amplification, Chlamydia
Medical Specialty	Microbiology
Product Code	MKZ
Device Class	1
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; K032194

This pre-market application is to clear the APTIMA Combo 2 Assay for use on the PANTHER System. The APTIMA Combo 2 Assay was cleared on the predicate TIGRIS System on December 24, 2003 (K032194). Clearance of this 510(k) will add the PANTHER System as an optional platform. No changes were introduced to the APTIMA Combo 2 Assay reagents.

2

Device Description

The APTIMA Combo 2 Assay combines the technologies of target capture, TMA, and DKA.

Specimens are collected and transferred into their respective specimen transport tubes. The transport solutions in these tubes release the rRNA targets and protect them from degradation during storage. When the APTIMA Combo 2 Assay is performed in the laboratory, the farget rRNA molecules are isolated from specimens by use of capture oligomers via target capture that utilizes magnetic microparticles. The capture oligomers contain sequences complementary to specific regions of the target molecules as well as a string of deoxyadenosine residues. A separate capture oligomer is used for each target. During the hybridization step, the sequence specific regions of the capture oligomers bind to specific regions of the target molecules. The capture oligomer:target complex is then captured out of solution by decreasing the temperature of the reaction to room temperature. This temperature reduction allows hybridization to occur between the deoxyadenosine region on the capture oligomer and the poly-deoxythymidine molecules that are covalently attached to the magnetic particles. The microparticles, including the captured target molecules bound to them, are pulled to the side of the reaction vessel using magnets and the supernatant is aspirated. The particles are washed to remove residual specimen matrix that may contain amplification reaction inhibitors. After the target capture steps are completed, the specimens are ready for amplification.

Target amplification assays are based on the ability of complementary oligonucleotide primers to specifically anneal and allow enzymatic amplification of the target nucleic acid strands. The APTIMA Combo 2 Assay replicates a specific region of the 23S rRNA from CT and a specific region of the 16S rRNA from GC via DNA intermediates. A unique set of primers is used for each target molecule. Detection of the rRNA amplification product sequences (amplicon) is achieved using nucleic acid hybridization. Single-stranded chemiluminescent DNA probes, which are complementary to a region of each target amplicon, are labeled with different acridinium ester molecules. The labeled DNA probes combine with amplicon to form stable RNA:DNA hybrids. The Selection Reagent differentiates hybridized from unhybridized probe, eliminating the generation of signal from unhybridized probe. During the detection step, light emitted from the labeled RNA:DNA hybrids is measured as photon signals in a luminometer,

3

and are reported as Relative Light Units (RLU). In DKA, differences in the kinetic profiles of the CT and GC labeled probes allow for the differentiation of signal; kinetic profiles are derived from measurements of photon output during the detection read time. The chemiluminescent detection for CT signal has very rapid kinetics and has the "flasher" kinetic type. The chemiluminescent detection reaction for GC signal is relatively slower and has the "glower" kinetic type. Assay results are determined by a cut-off based on the total RLU and the kinetic curve type.

Intended Use

The AC2 Assay intended use is stated below.

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) to aid in the diagnosis of chlamydial and/or gonococcal urogenital disease using the PANTHER System as specified.

On the PANTHER System, the assay may be used to test the following specimens from symptomatic and asymptomatic individuals: clinician-collected endocervical, vaginal and male urethral swab specimens, clinician-collected gynecological specimens collected in the PreservCyt Solution, and patient-collected vaginal swab specimens'.

'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.

4

Comparison to Predicate

: ·

· A comparison of the AC2 Assay on the PANTHER System with the predicate TIGRIS System is summarized below.

| Item | AC2 on TIGRIS System
(Predicate Device) | AC2 on PANTHER System |
|---------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Device Class | II | II |
| Regulation Specialty | Microbiology | Microbiology |
| Qualitative /Quantitative
Assay | Qualitative | Qualitative |
| Function | Detection and differentiation of
rRNA from Chlamydia
trachomatis and Neisseria
gonorrhoeae | Detection and differentiation of
rRNA from Chlamydia
trachomatis and Neisseria
gonorrhoeae |
| Indications For Use /
Intended Use | Detection and differentiation of
rRNA from Chlamydia
trachomatis and Neisseria
gonorrhoeae on the DTS and
TIGRIS Systems. | Detection and differentiation of
rRNA from Chlamydia
trachomatis and Neisseria
gonorrhoeae on the PANTHER
System. |
| Specimen Types | Female specimens:
• Vaginal swab
• Endocervical swab
• ThinPrep in PreservCyt
solution
• Urine
Male Specimens:
• Urethral Swab | Female specimens:
• Vaginal swab
• Endocervical swab
• ThinPrep in PreservCyt
solution
Male Specimens:
• Urethral Swab |
| Item | AC2 on TIGRIS System
(Predicate Device) | AC2 on PANTHER System |
| | Swabs:
After collection, transport and
store swab in transport tube at 2-
30°C and test within 60 days. If
longer storage is desired, freeze at
-20°C to -70°C for up to 365
days. | Swabs:
After collection, transport and
store swab in transport tube at 2-
30°C and test within 60 days. If
longer storage is desired, freeze at
-20°C to -70°C for up to 365
days. |
| Specimen
Transport/Storage | ThinPrep Liquid Pap in
PreservCyt:
Transport and store in PreservCyt
solution at 2-30°C for up to 30
days. After transfer to APTIMA
specimen transfer tube, store at
15-30°C for 14 days or store at
2-8°C for 30 days. If longer
storage is desired, freeze at -20°C
to -70°C for up to 365 days.
Urine:
After collection, transfer urine to
transport tube within 24 hours
and store at 2-30°C; Must be
assayed within 30 days after
transfer. If held longer, freeze -at
-20°C to -70°C for up to 365 days. | ThinPrep Liquid Pap in
PreservCyt:
Transport and store in PreservCyt
solution at 2-30°C for up to 30
days. After transfer to APTIMA
specimen transfer tube, store at
15-30°C for 14 days or store at
2-8°C for 30 days. If longer
storage is desired, freeze at -20°C
to -70°C for up to 365 days. |
| Type of Assay | Nucleic Acid Amplification Test | Nucleic Acid Amplification Test |
| Technology* | Target Capture (TC),
Transcription-Mediated
Amplification (TMA),
Hybridization Protection Assay .
(HPA) | Target Capture (TC),
Transcription-Mediated
Amplification (TMA),
Hybridization Protection Assay
(HPA) |
| Detection Format | HPA which provides relative
light units (RLUs) that are
assessed against an established
assay cutoff | HPA which provides relative light
units (RLUs) that are assessed
against an established assay
cutoff |

Comparison to Predicate Device

5

*A number of Gen-Probe APTIMA assays (that are based on TC, TMA, and HPA technologics) have been cleared by FDA including: the AC2 Assay (K060652), the APTIMA CT Assay (K061413) and the APTIMA GC Assay (K061509) for use on the automated TIGRIS System and the semiautomated DTS Systems.

6

Performance Data

Brief Description of Non-Clinical Data

The following studies were conducted to support analytical performance of the PANTHER System.

Analytical Sensitivity Study

Chlamydia trachomatis analytical sensitivity (limits of detection) was determined by directly comparing dilutions of CT organisms in the APTIMA COMBO 2 assay. The analytical sensitivity claim for the assay is 1 IFU/assay (7.25 IFU/swab, 9.75 IFU/mL, PreservCyt Solution liquid Pap). However, dilutions of less than 1 IFU/assay tested positive in the APTIMA COMBO 2 Assay (for this study, 100% positivity was observed in samples containing CT concentrations of 0.03 IFU/mL).

Neisseria gonorrhoeae analytical sensitivity (limits of detection) was determined by directly comparing dilutions of GC organismsin the APTIMA COMBO 2 Assay. The analytical sensitivity claim for the assay is 50 cells/assay (362 cells/swab, 488 cells/mL PreservCyt Solution liquid Pap). However, dilutions of less than 50 cells/assay tested positive in the APTIMA COMBO 2 Assay (for this study, 100% positivity was observed in samples containing GC concentrations of 0.04 CFU/mL).

Within Laboratory Precision Study

APTIMA COMBO 2 Assay precision was evaluated at Gen-Probe using the PANTHER System. Testing was performed using three PANTHER Systems and three lots of assay reagents. Testing was performed over 24 days.

Reproducibility panel members were created using negative PreservCyt Solution liquid Pap specimens and STM. The positive panel members were created by spiking CT and/or GC organisms to the targeted concentrations shown in Table 1.

7

.

For each panel member, Table 1 presents mean RLU, between-instrument, between-lot, between-run, within-run, and overall variation as SD and percent CV. Percent agreement with expected results is also shown.

Page 8 of 26

8

Table 1. PANTHER System Within Laboratory Precision Data

| | Target Concentration | Instruments | | Between
Lots | | Between
Runs | | Within
Runs | | Total | |--------|----------------------|---- | Matrix | CT
(IFU/mL) | GC
(CFU/mL) | Agreed/
N | Agrmt
(%) | RLU
(x1000) | SD
(x1000) | CV
(%) | SD
(x1000) | CV
(%) | SD
(x1000) | CV
(%) | SD
(x1000) | CV
(%) | SD
(x1000) | CV
(%) |
| STM | 0 | 0 | | 0.25 | 0 | | 2.5 | 0 | | 25 | 0 | | 0 | 12.5 | | 0 | 125 | | 0 | 1250 | | 2.5 | 125 | | 2.5 | 2500 | | 1000 | 2500 | | 1000 | 125 | PCyt | 0 | 0 | | 0.25 | 0 | | 2.5 | 0 | | 25 | 0 | | 0 | 12.5 | | 0 | 125 | | 0 | 1250 | | 2.5 | 125 | | | Mean | Between
|
------------|--------------|--------------|----------------|------------------------|-----------|-----------------|-----------|-----------------|-----------|----------------|-----------|---------------|-----------|
| 96/96 | 100 | 6 | 0.1 | 1.0 | 0.9 | 13.5 | 0.0 | 0.0 | 1.0 | 15.7 | 1.3 | 20.1 |
| 95/95 | 100 | 1226 | 70.0 | 5.7 | 20.0 | 1.6 | 8.4 | 0.7 | 47.1 | 3.8 | 87.1 | 7.1 |
| 96/96 | 100 | 1249 | 78.0 | 6.2 | 6.1 | 0.5 | 0.0 | 0.0 | 32.9 | 2.6 | 84.8 | 6.8 |
| 95/95 | 100 | 1268 | 72.9 | 5.7 | 15.3 | 1.2 | 0.0 | 0.0 | 39.6 | 3.1 | 84.3 | 6.6 |
| 96/96 | 100 | 1081 | 18.4 | 1.7 | 28.6 | 2.6 | 0.0 | 0.0 | 26.7 | 2.5 | 43.2 | 4.0 |
| 96/96 | 100 | 1266 | 29.8 | 2.4 | 0.0 | 0.0 | 8.9 | 0.7 | 27.6 | 2.2 | 41.6 | 3.3 |
| 96/96 | 100 | 1309 | 29.4 | 2.2 | 0.0 | 0.0 | 9.8 | 0.8 | 31.8 | 2.4 | 44.4 | 3.4 |
| 96/96 | 100 | 2456 | 86.6 | 3.5 | 0.0 | 0.0 | 0.0 | 0.0 | 53.0 | 2.2 | 101.5 | 4.1 |
| 96/96 | 100 | 2509 | 73.1 | 2.9 | 0.0 | 0.0 | 19.8 | 0.8 | 46.8 | 1.9 | 89.0 | 3.5 |
| 96/96 | 100 | 2496 | 31.7 | 1.3 | 6.1 | 0.2 | 0.0 | 0.0 | 193.7 | 7.8 | 196.3 | 7.9 |
| 96/96 | 100 | 2471 | 83.6 | 3.4 | 9.4 | 0.4 | 0.0 | 0.0 | 52.4 | 2.1 | 99.1 | 4.0 |
| 96/96 | 100 | 7 | 0.0 | 0.0 | 0.8 | 11.7 | 0.0 | 0.0 | 1.5 | 22.4 | 1.7 | 24.7 |
| 96/96 | 100 | 1113 | 92.3 | 8.3 | 30.1 | 2.7 | 0.0 | 0.0 | 63.6 | 5.7 | 116.0 | 10.4 |
| 96/96 | 100 | 1194 | 62.5 | 5.2 | 24.8 | 2.1 | 0.0 | 0.0 | 47.0 | 3.9 | 82.1 | 6.9 |
| 95/95 | 100 | 1222 | 65.1 | 5.3 | 26.4 | 2.2 | 14.7 | 1.2 | 35.0 | 2.9 | 79.8 | 6.5 |
| 93/93 | 100 | 994 | 33.3 | 3.3 | 36.9 | 3.7 | 16.0 | 1.6 | 26.2 | 2.6 | 58.4 | 5.9 |
| 95/95 | 100 | 1189 | 40.1 | 3.4 | 4.5 | 0.4 | 10.9 | 0.9 | 21.4 | 1.8 | 47.0 | 4.0 |
| 95/95 | 100 | 1239 | 37.7 | 3.0 | 7.5 | 0.6 | 13.6 | 1.1 | 18.0 | 1.5 | 44.6 | 3.6 |
| 95/95 | 100 | 2333 | 99.7 | 4.3 | 35.3 | 1.5 | 12.6 | 0.5 | 48.9 | 2.1 | 117.2 | 5.0 |

Agent = ageemen, CV = cellicient of variation, N = multion liggio = Pesserv, V.V = reblive light uni. 3D = standad deviation, STM = swab casport masjium.

Nee; Variability form some to reasonal the variability de to hose tures is very stall. Were this econs, the minimity so messeed with standed levision and 54.7 kset o ().

9

Carryover Studies for the PANTHER System

A multi-run analytical study was conducted using spiked panels on three PANTHER Systems. Carryover was assessed using approximately 20% high titer GC samples dispersed between negative samples. The runs included clusters of high positive samples with clusters of negative samples as well as single high positives dispersed in a specific pattern within the run. High titer samples were made using GC rRNA spiked into STM to give a final concentration equivalent to 2.5 x 108 CFU/mL. Testing was carried out using 5 runs on cach of three PANTHER Systems. Carryover was calculated from a total of 2938 valid negative results. The overall carryover rate was 0% with a 95% confidence interval of 0-0.1%.

10

Brief Description of Clinical Data

Prevalence

The prevalence of CT and GC in patient populations depends on risk factors such as age, gender, the presence or absence of symptoms, the type of clinic, and the sensitivity of the test used to detect infections. A summary of the prevalence of three CT and GC disease outcomes, as determined by the APTIMA COMBO 2 Assay on the PANTHER System in the clinical trial, is shown in Tables 2, 3, and 4 by specimen type and clinical site.

Table 2. Prevalence of CT+/GC- Infections as Determined by the APTIMA COMBO 2 Assay by Specimen Type and Clinical Site

	CT+/GC- Prevalence % (# positive/# tested with valid results)
Site	Male Urethral
Swab	Clinician-
Collected/Patient
-collected
Vaginal Swab	PreservCyt	Endocervical
Swab
1	0(-)	9.9 (21/212)	8.9 (20/225)	10.4 (20/193)
2	13.9 (28/202)	8.3 (19/230)	8.8 (21/239)	8.2 (19/231)
3	1.3 (1/76)	2.7 (6/222)	3.1 (7/226)	2.7 (6/223)
4	24.4 (33/135)	11.7 (40/342)	10.2 (35/342)	11.3 (38/337)
5	0(-)	4.5 (1/22)	4.8 (1/21)	4.3 (1/23)
6	21.5 (28/130)	11.9 (13/109)	8.7 (10/115)	8.8 (10/114)
7	16.7 (1/6)	3.2 (5/157)	2.5 (4/161)	2.6 (4/152)
All	16.6 (91/549)	8.1 (105/1294)	7.4 (98/1329)	7.7 (98/1273)

PreservCyt = PreservCyt Solution Liquid Pap

11

:

	CT-/GC+ Prevalence % (# positive/# tested with valid results)
Site	Male Urethral
Swab	Clinician-
Collected/Patient-
collected Vaginal
Swab	PreservCyt	Endocervical
Swab
1	0 ( - )	3.3 (7/212)	2.7 (6/225)	3.1 (6/193)
2	5.9 (12/202)	3.9 (9/230)	4.6 (11/239)	4.8 (11/231)
3	1.3 (1/76)	0.5 (1/222)	0.4 (1/226)	0.4 (1/223)
4	1.5 (2/135)	1.5 (5/342)	1.5 (5/342)	1.8 (6/337)
5	0 ( - )	0.0 (0/22)	0.0 (0/21)	0.0 (0/23)
6	5.4 (7/130)	3.7 (4/109)	1.7 (2/115)	1.8 (2/114)
7	0.0 (0/6)	2.5 (4/157)	2.5 (4/161)	2.6 (4/152)
All	4.0 (22/549)	2.3 (30/1294)	2.2 (29/1329)	2.4 (30/1273)

Table 3. Prevalence of CT-/GC+ Infections as Determined by the APTIMA COMBO 2 Assay by Specimen Type and Clinical Site

PreservCyt = PreservCyt Solution Liquid Pap

Table 4. Prevalence of CT+/GC+ Infections as Determined by the APTIMA COMBO 2 Assay by Specimen Type and Clinical Site

| Site | Male Urethral
Swab | CT+/GC+ Prevalence % (# positive/# tested with valid results)
Clinician-
Collected/Patient-
collected Vaginal
Swab | PreservCyt | Endocervical
Swab |
|------|-----------------------|--------------------------------------------------------------------------------------------------------------------------------|---------------|----------------------|
| | | | | |
| 2 | 3.0 (6/202) | 1.3 (3/230) | 0.8 (2/239) | 0.9 (2/231) |
| 3 | 0.0 (0/76) | 0.0 (0/222) | 0.0 (0/226) | 0.0 (0/223) |
| 4 | 4.4 (6/135) | 1.2 (4/342) | 0.9 (3/342) | 0.9 (3/337) |
| 5 | 0 (-) | 0.0 (0/22) | 0.0 (0/21) | 0.0 (0/23) |
| 6 | 0.8 (1/130) | 0.9 (1/109) | 0.9 (1/115) | 0.9 (1/114) |
| 7 | 0.0 (0/6) | 0.6 (1/157) | 0.6 (1/161) | 0.7 (1/152) |
| All | 2.4 (13/549) | 1.3 (17/1294) | 1.1 (14/1329) | 1.1 (14/1273) |

PreservCyt = PreservCyt Solution Liquid Pap

12

Positive and Negative Predictive Values for Hypothetical Prevalence Rates

The estimated positive and negative predictive values (PPV and NPV) of the APTIMA COMBO 2 Assay for different hypothetical prevalence rates are shown for each specimen type in Table 5. The PPV and NPV are derived for different hypothetical prevalence rates using the sensitivity and specificity estimates for each specimen type from the clinical performance study (see Table 6 and Table 9).

| Specimen Type | Hypothetical
Prevalence (%) | CT Detection | | GC Detection | |
|-------------------------------------------------------------------------------|--------------------------------|--------------|---------|--------------|---------|
| | | PPV (%) | NPV (%) | PPV (%) | NPV (%) |
| Male Urethral
Swab | 1 | 53.1 | 100 | 100 | 100 |
| | 2 | 69.6 | 100 | 100 | 100 |
| | 5 | 85.5 | 100 | 100 | 100 |
| | 10 | 92.6 | 100 | 100 | 100 |
| | 15 | 95.2 | 100 | 100 | 100 |
| | 20 | 96.6 | 100 | 100 | 100 |
| | 25 | 97.4 | 100 | 100 | 100 |
| Clinician-
Collected Vaginal
Swab/
Patient-Collected
Vaginal Swab | 1 | 38.9 | 100 | 70.6 | 100 |
| | 2 | 56.3 | 99.9 | 82.9 | 100 |
| | 5 | 76.8 | 99.9 | 92.6 | 99.9 |
| | 10 | 87.5 | 99.7 | 96.3 | 99.7 |
| | 15 | 91.7 | 99.5 | 97.7 | 99.6 |
| | 20 | 94.0 | 99.3 | 98.3 | 99.4 |
| | 25 | 95.5 | 99.1 | 98.8 | 99.2 |
| PreservCyt
Solution Liquid
Pap | 1 | 100 | 100 | 100 | 100 |
| | 2 | 100 | 100 | 100 | 100 |
| | 5 | 100 | 99.9 | 100 | 100 |
| | 10 | 100 | 99.8 | 100 | 100 |
| | 15 | 100 | 99.7 | 100 | 100 |
| | 20 | 100 | 99.6 | 100 | 100 |
| | 25 | 100 | 99.4 | 100 | 100 |
| Endocervical
Swab | 1 | 58.5 | 100 | 85.8 | 100 |
| | 2 | 74.0 | 99.9 | 92.4 | 100 |
| | 5 | 88.0 | 99.9 | 96.9 | 100 |
| | 10 | 93.9 | 99.7 | 98.5 | 100 |
| | 15 | 96.1 | 99.5 | 99.1 | 100 |
| | 20 | 97.2 | 99.3 | 99.3 | 100 |
| | 25 | 97.9 | 99.1 | 99.5 | 100 |

Table 5. Positive and Negative Predictive Values for Hypothetical Prevalence Rates by Specimen Type

13

Clinical Study Results

A prospective, multicenter clinical study was conducted to establish the performance characteristics of the APTIMA COMBO 2 Assay on the PANTHER System. Specimens were collected from symptomatic and asymptomatic men (n=580) and women (n=1332) enrolled from 7 geographically and ethnically diverse US clinical sites, including obstetrics and gynecology, family planning, public health, and STD clinics. Subjects were classified as symptomatic if symptoms were reported by the subject. Subjects were classified as asymptomatic if the subject did not report symptoms. Of the 580 male subjects, none were 25 years of age. Of the 1332 female subjects, 11 were 14 to 15 years of age, 59 were 16 to 17 years of age, 319 were 18 to 20 years of age, 401 were 21 to 25 years of age, and 542 were >25 years of age.

Up to 2 specimens were collected from each male subject (1 urethral swab and 1 first-catch urine, in that order) and up to 4 specimens were collected from each female subject (1 first-catch urine, 1 vaginal swab, 1 PreservCyt Solution liquid Pap specimen, and 1 endocervical swab, in that order). All specimens were clinician-collected except urine specimens and approximately half of the vaginal swab specimens, which were collected by the subject at the clinic. Approximately half of the PreservCyt Solution liquid Pap specimens were collected with a broom-type device and half were collected with a spatula and cytobrush. Samples were prepared for APTIMA testing in accordance with the appropriate APTIMA specimen collection kit package insert instructions.

All evaluable samples (567 male urethral swab, 1319 vaginal swab, 1330 PreservCyt, and 1310 endocervical swab samples) were tested with the APTIMA COMBO 2 Assay on the PANTHER System in accordance with package insert instructions. The samples were split amongst three laboratories (two external laboratories and GEN-PROBE). Samples with initial invalid, equivocal, or error results were retested. Eighteen (18) male urethral swab, 25 vaginal swab, 1 PreservCyt, and 37 endocervical swab specimens had final invalid results and were excluded from the analyses. Most of the invalid results were due to insufficient sample volume. One vaginal swab and 1 endocervical swab had final CT equivocal results and 1 PreservCyt and 1 endocervical swab had final GC equivocal results and were excluded from the analyses.

14

Male urethral swab, male and female urine, and PreservCyt samples were tested with cleared nucleic acid amplification tests (NAATs) to establish the infected status. The infected status algorithm used results from two specimen types and two reference NAATs. Subjects were categorized as infected if a positive result occurred in each of the 2 reference NAATs. For female subjects, if the positive NAAT results occurred only in the urine specimens and not in the PreservCyt specimens, the subject was categorized as infected; however, for the evaluation of the non-urine specimen types, the specimens were considered non-infected. Subjects that could not be categorized as infected or not infected were excluded from the performance analyses.

Chlamydia trachomatis Performance Results

Performance characteristics of the APTIMA COMBO 2 Assay were estimated by comparing PANTHER System results to the infected status algorithm. Table 6 shows the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the APTIMA COMBO 2 Assay for CT detection and the prevalence of CT (based on the infected status) in each specimen type.

| Spec
Type | n | TP | FP | TN | FN | Prev % | Sensitivity %
(95% CI)¹ | Specificity %
(95% CI)¹ | PPV %
(95% CI)² | NPV %
(95% CI)² |
|--------------|------|-----|----|------|----|--------|----------------------------|----------------------------|---------------------|---------------------|
| MS | 549 | 100 | 4 | 445 | 0 | 18.2 | 100
(96.3-100) | 99.1
(97.7-99.7) | 96.2
(90.8-98.9) | 100
(99.2-100) |
| CVS/
PVS | 1274 | 104 | 18 | 1149 | 3 | 8.4 | 97.2
(92.1-99.0) | 98.5
(97.6-99.0) | 85.2
(78.8-90.5) | 99.7
(99.3-99.9) |
| PCyt | 1311 | 112 | 0 | 1197 | 2 | 8.7 | 98.2
(93.8-99.5) | 100
(99.7-100) | 100
(96.9-100) | 99.8
(99.4-100) |
| FS | 1254 | 104 | 8 | 1139 | 3 | 8.5 | 97.2
(92.1-99.0) | 99.3
(98.6-99.6) | 92.9
(87.1-96.7) | 99.7
(99.3-99.9) |

Table 6. Performance Characteristics of the APTIMA COMBO 2 Assay for CT Detection

CI = confidence interval, CVS = clinician-collected vaginal swab, FN = false negative, FP = false positive, FS = female endocervical swab, MS = male urethral swab, NPV = negative predictive value. PCyt = PreservCyt Solution liquid Pap, PV = positive predictive value, Prev = prevalent-collected vaginal swab, Spec = specimen, TN = true negative. TY = true positive.

' Score Cl

2 PPV 95% CI computed from the exact 95% CI for the positive likelihood ratio, NPV 95% CI computed from the exact 95% CI from the negative likelihood ratio.

Table 7 shows the sensitivity, specificity, PPV, and NPV of the APTIMA COMBO 2 Assay for CT detection and the prevalence of CT (based on the infected status) in each specimen type by symptom status. CT prevalence was higher in symptomatic men and women.

15

| Spec
Type | Sx
Status | n | TP | FP | TN | FN | Prev
% | Sensitivity
%
(95% CI)¹ | Specificity
%
(95% CI)¹ | PPV %
(95% CI)² | NPV %
(95% CI)² |
|--------------|--------------|-----|----|----|-----|----|-----------|-------------------------------|-------------------------------|---------------------|---------------------|
| MS | Sym | 238 | 59 | 1 | 178 | 0 | 24.8 | 100
(93.9-100) | 99.4
(96.9-99.9) | 98.3
(91.5-100) | 100
(98.0-100) |
| | Asym | 311 | 41 | 3 | 267 | 0 | 13.2 | 100
(91.4-100) | 98.9
(96.8-99.6) | 93.2
(82.5-98.5) | 100
(98.7-100) |
| CVS/
PVS | Sym | 810 | 73 | 8 | 729 | 0 | 9.0 | 100
(95.0-100) | 98.9
(97.9-99.4) | 90.1
(82.3-95.5) | 100
(99.5-100) |
| | Asym | 464 | 31 | 10 | 420 | 3 | 7.3 | 91.2
(77.0-97.0) | 97.7
(95.8-98.7) | 75.6
(63.1-86.2) | 99.3
(98.1-99.8) |
| PCyt | Sym | 838 | 76 | 0 | 762 | 0 | 9.1 | 100
(95.2-100) | 100
(99.5-100) | 100
(95.4-100) | 100
(99.5-100) |
| | Asym | 473 | 36 | 0 | 435 | 2 | 8.0 | 94.7
(82.7-98.5) | 100
(99.1-100) | 100
(91.1-100) | 99.5
(98.5-99.9) |
| FS | Sym | 794 | 71 | 5 | 718 | 0 | 8.9 | 100
(94.9-100) | 99.3
(98.4-99.7) | 93.4
(85.9-97.8) | 100
(99.5-100) |
| | Asym | 460 | 33 | 3 | 421 | 3 | 7.8 | 91.7
(78.2-97.1) | 99.3
(97.9-99.8) | 91.7
(79.9-98.0) | 99.3
(98.1-99.8) |

Table 7. Performance Characteristics of the APTIMA COMBO 2 Assay for CT Detection by Symptom Status

Asym = asymplomatic, CI = confidence interval, CVS = clinician-collected vaginal swab, FN = false positive, FS = female endocervical swab, MS = male urettral swab, NPV = negative predictive value, PCyt = PreservCyt Solution liquid Pap, PV = positive predictive value, Prev = prevalent-collected vaginal swab, Spec = specimen, Sx = symplom, Sym = symplomatic, TN = true negative, TP = true positive.

l Score Cl

2 PPV 95% Cl computed from the exact 95% Cl for the positive likelihood ratio, NPV 95% Cl computed from the negative likelihood ratio.

Table 8 shows the sensitivity, specificity, PPV, and NPV of the APTIMA COMBO 2 Assay for CT detection and the prevalence of CT (based on the infected status) in each specimen type by clinical site. CT prevalence varied across clinical sites, as expected.

16

Table 8. Performance Characteristics of the APTIMA COMBO 2 Assay for CT Detection by
Clinical Site

| Spec
Type | Site | n | TP | FP | TN | FN | Prev
% | Sensitivity %
(95% CI)¹ | Specificity %
(95% CI)¹ | PPV %
(95% CI)² | NPV %
(95% CI)² |
|--------------|------|-----|----|----|-----|----|-----------|----------------------------|----------------------------|---------------------|---------------------|
| MS | 1 | 0 | 0 | 0 | 0 | 0 | NC | NC | NC | NC | NC |
| | 2 | 202 | 34 | 0 | 168 | 0 | 16.8 | 100
(89.8-100) | 100
(97.8-100) | 100
(90.3-100) | 100
(98.0-100) |
| | 3 | 76 | 0 | 1 | 75 | 0 | 0.0 | NC | 98.7
(92.9-99.8) | 0.0
(NC) | 100
(NC) |
| | 4 | 135 | 36 | 3 | 96 | 0 | 26.7 | 100
(90.4-100) | 97.0
(91.5-99.0) | 92.3
(80.9-98.3) | 100
(96.6-100) |
| | 5 | 0 | 0 | 0 | 0 | 0 | NC | NC | NC | NC | NC |
| | 6 | 130 | 29 | 0 | 101 | 0 | 22.3 | 100
(88.3-100) | 100
(96.3-100) | 100
(88.9-100) | 100
(96.7-100) |
| | 7 | 6 | 1 | 0 | 5 | 0 | 16.7 | 100
(20.7-100) | 100
(56.6-100) | 100
(6.9-100) | 100
(80.4-100) |
| CVS/
PVS | 1 | 211 | 23 | 6 | 182 | 0 | 10.9 | 100
(85.7-100) | 96.8
(93.2-98.5) | 79.3
(64.2-91.2) | 100
(98.2-100) |
| | 2 | 230 | 18 | 4 | 206 | 2 | 8.7 | 90.0
(69.9-97.2) | 98.1
(95.2-99.3) | 81.8
(64.7-93.9) | 99.0
(97.1-99.9) |
| | 3 | 213 | 6 | 0 | 206 | 1 | 3.3 | 85.7
(48.7-97.4) | 100
(98.2-100) | 100
(64.1-100) | 99.5
(98.1-100) |
| | 4 | 335 | 40 | 4 | 291 | 0 | 11.9 | 100
(91.2-100) | 98.6
(96.6-99.5) | 90.9
(79.8-97.3) | 100
(98.8-100) |
| | 5 | 21 | 1 | 0 | 20 | 0 | 4.8 | 100
(20.7-100) | 100
(83.9-100) | 100
(6.5-100) | 100
(95.3-100) |
| | 6 | 107 | 11 | 3 | 93 | 0 | 10.3 | 100
(74.1-100) | 96.9
(91.2-98.9) | 78.6
(56.4-94.6) | 100
(96.8-100) |
| | 7 | 157 | 5 | 1 | 151 | 0 | 3.2 | 100
(56.6-100) | 99.3
(96.4-99.9) | 83.3
(47.4-99.5) | 100
(98.3-100) |
| PCyt | 1 | 225 | 27 | 0 | 198 | 0 | 12.0 | 100
(87.5-100) | 100
(98.1-100) | 100
(88.1-100) | 100
(98.3-100) |
| | 2 | 239 | 23 | 0 | 216 | 0 | 9.6 | 100
(85.7-100) | 100
(98.3-100) | 100
(86.3-100) | 100
(98.4-100) |
| | 3 | 217 | 7 | 0 | 210 | 0 | 3.2 | 100
(64.6-100) | 100
(98.2-100) | 100
(65.7-100) | 100
(98.7-100) |
| | 4 | 335 | 38 | 0 | 295 | 2 | 11.9 | 95.0
(83.5-98.6) | 100
(98.7-100) | 100
(91.5-100) | 99.3
(97.8-99.9) |
| | 5 | 21 | 1 | 0 | 20 | 0 | 4.8 | 100
(20.7-100) | 100
(83.9-100) | 100
(6.5-100) | 100
(95.3-100) |
| | 6 | 113 | 11 | 0 | 102 | 0 | 9.7 | 100
(74.1-100) | 100
(96.4-100) | 100
(75.2-100) | 100
(97.0-100) |
| | 7 | 161 | 5 | 0 | 156 | 0 | 3.1 | 100
(56.6-100) | 100
(97.6-100) | 100
(57.8-100) | 100
(98.4-100) |

Cl = confidence interval, CVS = clinician-collected vaginal swab, FN = false positive, FS = female endocervical swab, MS = male urethral swah, NPV = negative value, PCy1 = PreservCyt Solution liquid Pap, PPV = postive predictive value, Prev = prevalence, PVS = patient-collected vaginal swab, Spec = specimen, TN = true negative, TP = true positive. ' Score Cl

2 PPV 95% Cl computed from the exact 95% Cl for the positive likelihood ratio, NPV 95% Cl computed from the cract 95% Cl from the negative likelihood ratio.

17

| Spec
Type | Site | n | TP | FP | TN | FN | Prev % | Sensitivity %
(95% CI)¹ | Specificity %
(95% CI)¹ | PPV %
(95% CI)² | NPV %
(95% CI)² |
|--------------|------|-----|----|----|-----|----|--------|----------------------------|----------------------------|---------------------|--------------------|
| FS | 1 | 193 | 24 | 3 | 166 | 0 | 12.4 | 100
(86.2-100) | 98.2
(94.9-99.4) | 88.9
(73.6-97.5) | 100
(98.0-100) |
| | 2 | 231 | 19 | 2 | 209 | 1 | 8.7 | 95.0
(76.4-99.1) | 99.1
(96.6-99.7) | 90.5
(73.4-98.6) | 99.5
(97.7-100) |
| | 3 | 214 | 6 | 0 | 207 | 1 | 3.3 | 85.7
(48.7-97.4) | 100
(98.2-100) | 100
(64.1-100) | 99.5
(98.1-100) |
| | 4 | 330 | 39 | 2 | 288 | 1 | 12.1 | 97.5
(87.1-99.6) | 99.3
(97.5-99.8) | 95.1
(84.8-99.3) | 99.7
(98.2-100) |
| | 5 | 22 | 1 | 0 | 21 | 0 | 4.5 | 100
(20.7-100) | 100
(84.5-100) | 100
(6.5-100) | 100
(95.5-100) |
| | 6 | 112 | 11 | 0 | 101 | 0 | 9.8 | 100
(74.1-100) | 100
(96.3-100) | 100
(75.2-100) | 100
(97.0-100) |
| | 7 | 152 | 4 | 1 | 147 | 0 | 2.6 | 100
(51.0-100) | 99.3
(96.3-99.9) | 80.0
(40.0-99.4) | 100
(98.4-100) |

Table 8. Performance Characteristics of the APTIMA COMBO 2 Assay for CT Detection by Clinical Site (continued)

Cl = confidence interval, CVS = clinician-collected vaginal swab, FN = false positive, FS = female endocervical swab, MS = male urethral swab, NPV = negative predictive value, PCyt = PreservCyt Solution liquid Pap, PPV = positive predictive value, Prev = prevalence, PVS = patient-collected vaginal swab, Spec = specimen, TN = true negative, TP = true positive. 1 Score Cl

2 YPV 95% Cl computed from the exact 95% Cl for the positive likelihood ratio, NPV 95% Cl computed from the exact 95% Cl from the negative likelihood ratio.

Table 9 shows the sensitivity, specificity, PPV, and NPV of the APTIMA COMBO 2 Assay for GC detection and the prevalence of GC (based on the infected status) in each specimen type.

| Spec
Type | n | TP | FP | TN | FN | Prev
% | Sensitivity %
(95% CI)1 | Specificity %
(95% CI)1 | PPV %
(95% CI)2 | NPV %
(95% CI)2 |
|--------------|------|----|----|------|----|-----------|----------------------------|----------------------------|---------------------|--------------------|
| MS | 546 | 34 | 0 | 512 | 0 | 6.2 | 100
(89.8-100) | 100
(99.3-100) | 100
(90.2-100) | 100
(99.3-100) |
| VS | 1258 | 42 | 5 | 1210 | 1 | 3.4 | 97.7
(87.9-99.6) | 99.6
(99.0-99.8) | 89.4
(78.6-96.1) | 99.9
(99.6-100) |
| PCyt | 1293 | 43 | 0 | 1250 | 0 | 3.3 | 100
(91.8-100) | 100
(99.7-100) | 100
(92.1-100) | 100
(99.7-100) |
| FS | 1238 | 42 | 2 | 1194 | 0 | 3.4 | 100
(91.6-100) | 99.8
(99.4-100) | 95.5
(85.4-99.4) | 100
(99.7-100) |

Table 9. Performance Characteristics of the APTIMA COMBO 2 Assay for GC Detection

Cl = confidence interval, CVS = clinician-collected vaginal swab, FN = false positive, FS = female endocervical swab, MS = male urethral swab, NPV = negative predictive value, PCyt = PreservCyt Solution liquid Pap, PV = positive predictive value, Prev = prevalence, PVS = patient-collected vaginal swab, Spec = specimen, TN = true negative, TV = true positive.

Score CI

2 PPV 95% Cl computed from the exact 95% Cl for the positive likelihood ratio, NPV 95% Cl computed from the exact 95% Cl from the negative likelihood ratio.

18

Table 10 shows the sensitivity, specificity, PPV, and NPV of the APTIMA COMBO 2 Assay for GC detection and the prevalence of GC (based on the infected status) in each specimen type by symptom status. GC prevalence was higher in symptomatic men but similar in symptomatic and asymptomatic women.

Table 10. Performance Characteristics of the APTIMA COMBO 2 Assay for GC Detection by Symptom Status

| Spec
Type | Sx
Status | n | TP | FP | TN | FN | Prev
% | Sensitivity %
(95% CI)¹ | Specificity %
(95% CI)¹ | PPV %
(95% CI)² | NPV %
(95% CI)² |
|--------------------|--------------|-----|----|----|-----|----|-----------|----------------------------|----------------------------|---------------------|--------------------|
| MS | Sym | 236 | 31 | 0 | 205 | 0 | 13.1 | 100
(89.0-100) | 100
(98.2-100) | 100
(89.5-100) | 100
(98.3-100) |
| MS | Asym | 310 | 3 | 0 | 307 | 0 | 1.0 | 100
(43.9-100) | 100
(98.8-100) | 100
(44.4-100) | 100
(99.3-100) |
| CVS/
PVS | Sym | 802 | 27 | 4 | 771 | 0 | 3.4 | 100
(87.5-100) | 99.5 (98.7-
99.8) | 87.1
(72.6-96.1) | 100
(99.6-100) |
| CVS/
PVS | Asym | 456 | 15 | 1 | 439 | 1 | 3.5 | 93.8
(71.7-98.9) | 99.8 (98.7-
100) | 93.8
(74.0-99.8) | 99.8
(98.9-100) |
| PCyt | Sym | 829 | 27 | 0 | 802 | 0 | 3.3 | 100
(87.5-100) | 100
(99.5-100) | 100
(88.0-100) | 100
(99.6-100) |
| PCyt | Asym | 464 | 16 | 0 | 448 | 0 | 3.4 | 100
(80.6-100) | 100
(99.1-100) | 100
(81.3-100) | 100
(99.3-100) |
| FS | Sym | 785 | 26 | 1 | 758 | 0 | 3.3 | 100
(87.1-100) | 99.9
(99.3-100) | 96.3
(82.4-99.9) | 100
(99.5-100) |
| FS | Asym | 453 | 16 | 1 | 436 | 0 | 3.5 | 100
(80.6-100) | 99.8
(98.7-100) | 94.1
(74.3-99.8) | 100
(99.3-100) |
| Spec
Type | Site | n | TP | FP | TN | FN | Prev
% | Sensitivity %
(95% CI)¹ | Specificity %
(95% CI)¹ | PPV %
(95% CI)² | NPV %
(95% CI)² |
| MS | 1 | 0 | 0 | 0 | 0 | 0 | NC | NC | NC | NC | NC |
| | 2 | 201 | 18 | 0 | 183 | 0 | 9.0 | 100
(82.4-100) | 100
(97.9-100) | 100
(83.1-100) | 100
(98.2-100) |
| | 3 | 76 | 1 | 0 | 75 | 0 | 1.3 | 100
(20.7-100) | 100
(95.1-100) | 100
(6.4-100) | 100
(98.7-100) |
| | 4 | 135 | 8 | 0 | 127 | 0 | 5.9 | 100
(67.6-100) | 100
(97.1-100) | 100
(68.8-100) | 100
(97.7-100) |
| | 5 | 0 | 0 | 0 | 0 | 0 | NC | NC | NC | NC | NC |
| | 6 | 128 | 7 | 0 | 121 | 0 | 5.5 | 100
(64.6-100) | 100
(96.9-100) | 100
(65.8-100) | 100
(97.7-100) |
| | 7 | 6 | 0 | 0 | 6 | 0 | 0.0 | NC | 100
(61.0-100) | NC | 100
( NC ) |
| CVS/
PVS | 1 | 207 | 13 | 2 | 192 | 0 | 6.3 | 100
(77.2-100) | 99.0
(96.3-99.7) | 86.7
(64.6-98.3) | 100
(98.4-100) |
| | 2 | 230 | 12 | 0 | 217 | 1 | 5.7 | 92.3
(66.7-98.6) | 100
(98.3-100) | 100
(77.8-100) | 99.5
(97.9-100) |
| | 3 | 206 | 1 | 0 | 205 | 0 | 0.5 | 100
(20.7-100) | 100
(98.2-100) | 100
(6.4-100) | 100
(99.5-100) |
| | 4 | 331 | 8 | 1 | 322 | 0 | 2.4 | 100
(67.6-100) | 99.7
(98.3-99.9) | 88.9
(59.1-99.7) | 100
(99.1-100) |
| | 5 | 21 | 0 | 0 | 21 | 0 | 0.0 | NC | 100
(84.5-100) | NC | 100
( NC ) |
| | 6 | 106 | 3 | 2 | 101 | 0 | 2.8 | 100
(43.9-100) | 98.1
(93.2-99.5) | 60.0
(24.6-92.8) | 100
(98.0-100) |
| | 7 | 157 | 5 | 0 | 152 | 0 | 3.2 | 100
(56.6-100) | 100
(97.5-100) | 100
(57.8-100) | 100
(98.3-100) |
| PCyt | 1 | 220 | 13 | 0 | 207 | 0 | 5.9 | 100
(77.2-100) | 100
(98.2-100) | 100
(78.0-100) | 100
(98.5-100) |
| | 2 | 239 | 13 | 0 | 226 | 0 | 5.4 | 100
(77.2-100) | 100
(98.3-100) | 100
(78.0-100) | 100
(98.6-100) |
| | 3 | 210 | 1 | 0 | 209 | 0 | 0.5 | 100
(20.7-100) | 100
(98.2-100) | 100
(6.4-100) | 100
(99.5-100) |
| | 4 | 331 | 8 | 0 | 323 | 0 | 2.4 | 100
(67.6-100) | 100
(98.8-100) | 100
(68.6-100) | 100
(99.1-100) |
| | 5 | 21 | 0 | 0 | 21 | 0 | 0.0 | NC | 100
(84.5-100) | NC | 100
( NC ) |
| | 6 | 111 | 3 | 0 | 108 | 0 | 2.7 | 100
(43.9-100) | 100
(96.6-100) | 100
(44.6-100) | 100
(98.1-100) |
| | 7 | 161 | 5 | 0 | 156 | 0 | 3.1 | 100
(56.6-100) | 100
(97.6-100) | 100
(57.8-100) | 100
(98.4-100) |
| Spec
Type
FS | Site | n | TP | FP | TN | FN | Prev
% | Sensitivity %
(95% CI)¹ | Specificity %
(95% CI)¹ | PPV %
(95% CI)² | NPV %
(95% CI)² |
| 1 | 1 | 189 | 12 | 1 | 176 | 0 | 6.3 | 100
(75.8-100) | 99.4
(96.9-99.9) | 92.3
(68.6-99.8) | 100
(98.2-100) |
| 2 | 2 | 231 | 13 | 0 | 218 | 0 | 5.6 | 100
(77.2-100) | 100
(98.3-100) | 100
(78.0-100) | 100
(98.5-100) |
| 3 | 3 | 207 | 1 | 0 | 206 | 0 | 0.5 | 100
(20.7-100) | 100
(98.2-100) | 100
(6.4-100) | 100
(99.5-100) |
| 4 | 4 | 327 | 8 | 1 | 318 | 0 | 2.4 | 100
(67.6-100) | 99.7
(98.2-99.9) | 88.9
(59.1-99.7) | 100
(99.1-100) |
| 5 | 5 | 22 | 0 | 0 | 22 | 0 | 0.0 | NC | 100
(85.1-100) | NC | 100
(NC) |
| 6 | 6 | 110 | 3 | 0 | 107 | 0 | 2.7 | 100
(43.9-100) | 100
(96.5-100) | 100
(44.6-100) | 100
(98.1-100) |
| 7 | 7 | 152 | 5 | 0 | 147 | 0 | 3.3 | 100
(56.6-100) | 100
(97.5-100) | 100
(57.9-100) | 100
(98.3-100) |

Asym = asymptomatic, CI = confidence interval, CVS = clinician-collected vaginal swab, FN = false negative, FP = false positive, FS = female endocervical swab, NPV = negative predictive value, PCy = PreservCyt Solution liquid Pap, PV = positive value, Prev = prevalence, PVS = patient-collected vaginal swab, Spec = specimen, Sx = symptom, Sym = symptomatic, TN = true negative, TP = true positive.

Score Cl

2 PPV 95% CI computed from the exact 95% CI for the positive likelihood ratio, NPV 95% CI computed from the exact 95% CI from the negative likelihood ratio.

Table 11 shows the sensitivity, specificity, PPV, and NPV of the APTIMA COMBO 2 Assay for GC detection and the prevalence of GC (based on the infected status) in each specimen type by clinical site for the clinical performance study. GC prevalence varied across clinical sites, as expected.

19

Table 11. Performance Characteristics of the APTIMA COMBO 2 Assay for GC Detection by Clinical Site

Cl = confidence interval, CVS = clinician-collected vaginal swab, FN = false positive, FS = female endocervical swab, MS = male urethral swab, NPV = negative predictive value, PCyt = PreservCyt Solution liquid Pap, PPV = positive predictive value, Prev = prevalent-collected vaginal swab, Spcc = specimen, TN = true negative, TV = true positive.

' Score CI

2 PPV 95% Cl computed from the exact 95% Cl for the positive likelihood ratio, NPV 95% Cl computed from the exact 95% Cl from the negative likelihood ratio.

20

Table 11. Performance Characteristics of the APTIMA COMBO 2 Assay for GC Detection by Clinical Site (continued)

CI = confidence interval, CVS = clinician-collected vaginal swab, FN = false negative, FP = false positive, FS = female endocervical swab, MS = male urethral swab, NPV = negative predictive value, PCyt = PreservCyt Solution liquid Pap, PPV = positive predictive value, Prev = prevalence, PVS = patient-collected vaginal swab, Spec = specimen, TN = 1rue negative, TV = true positive.

Score CI

2 PPV 95% Cl computed from the exact 95% CI for the positive likelihood ratio, NPV 95% CI computed from the exact 95% Cl from the negative likelihood ratio.

The frequency of test outcomes from reference NAAT and investigational PANTHER System testing is summarized in Table 13 for CT and in Tables 16 and Tables 16 and Table 15 for GC.

21

.

		Assay Results
		AC2 DTS	ACT TIGRIS	AC2
PANTHER	Symptom Status
CT Infected Status	MS	MU	MS	MU	MS	Sym	Asym
Infected	+	+	+	+	+	50	37
Infected	+	+	+	+	NA	4	1
Infected	+	+	+	-	+	2	0
Infected	+	+	+	+	+	4	2
Infected	+	-	+	-	+	3	2
Not Infected	+	+	-	-	-	0	1
Not Infected	+	-	-	-	+	0	1
Not Infected	+	-	-	-	-	1	1
Not Infected	-	-	+	-	-	3	2
Not Infected	-	-	-	+	-	1	1
Not Infected	-	-	-	-	+	1	2
Not Infected	-	-	-	-	-	173	262
Not Infected	-	-	-	-	NA	10	9
Not Infected	NA	-	-	-	NA	1	2

Table 12. CT Infected Status for Performance Evaluation in Male Urethral Swab Specimens

AC2 = APTIMA COMBO 2, ACT = APTIMA CT Assuy, Asym = asymptomatic, DTS = DTS Systems, MS = male urethral swab, MU = male urine, NA = result not available, PANTHER System, Sym = symptomatic, TIGRIS = TIGRIS DTS System

. .

:

.

:

.

22

				Assay Results
CT Infected
Status	AC2 TIGRIS		ACT
TIGRIS			AC2
PANTHER		Symptom Status
	PCyt	FU	PCyt	FU	CVS/PVS	PCyt	FS	Sym	Asym
Infected	+	+	+	+	+	+	+	62	26
Infected	+	+	+	+	+	+	+	0	1
Infected	+	+	+	+	+	+	NA	3	0
Infected	+	+	+	+	+	+	+	0	2
Infected	+	+	+	+	+	+	+	0	1
Infected	+	+	+	+	NA	+	+	1	1
Infected	+	+	+	+	NA	+	NA	2	1
Infected	+	+	+	+	+	+	+	4	1
Infected	+	-	+	+	NA	+	NA	0	1
Infected	+	-	+	+	+	+	+	4	0
Infected	+	-	+	+	+	+	+	0	1
Infected	+	NA	+	NA	+	+	+	0	1
Infected	+	NA	+	NA	+	+	+	0	1
Infected	+	NA	+	NA	+	+	+	0	1
Infected¹	-	+	-	+	+	-	+	1	1
Infected¹	-	+	-	+	+	-	-	2	0
Infected¹	-	+	-	+	+	-	+	1	1
Not Infected	+	-	-	-	-	-	+	0	2
Not Infected	-	+	-	-	-	-	+	1	0
Not Infected	-	-	+	-	+	-	+	0	1
Not Infected	-	-	+	-	+	-	-	5	0
Not Infected	-	-	-	+	+	-	+	0	1
Not Infected	-	-	-	+	+	-	NA	0	1
Not Infected	-	-	-	+	-	-	+	1	3
Not Infected	-	-	-	-	+	-	+	1	0
Not Infected	-	-	-	-	+	-	-	2	7
Not Infected	-	-	-	-	+	-	NA	2	0
Not Infected	-	-	-	-	+	-	+	2	2
Not Infected	-	-	-	-	+	-	-	680	396
Not Infected	-	-	-	-	-	-	NA	29	8
Not Infected	-	-	-	-	+	NA	-	1	0
Not Infected	-	-	-	-	NA	+	-	17	4
Not Infected	-	-	-	-	NA	-	NA	8	1
Not Infected	-	NA	-	-	-	-	-	8	6
Not Infected	-	NA	-	-	-	-	NA	0	1
Not Infected	NA	-	-	-	+	1	-	0	1
Not Infected	NA	-	-	-	-	-	NA	1	0
Not Infected	NA	-	-	-	NA	-	+	1	0

Table 13. CT Infected Status for Performance Evaluation in Female Vaginal Swab, PreservCyt Solution Liquid Pap, and Endocervical Swab Specimens

AC2 = APTIMA COMBO 2, ACT = APTIMA CT Assoy, Asym = asymplomatic, CVS = clinician-collected vaginal swab, FS = female endocervical swab, FU = female urine, NA = result not available, I'ANTHER System, PCy = PreservCyt Solution liquid Pap. I'VS

= patient-collected vaginal swab, Sym = symptomatic, TIGRIS = TIGRIS DTS System

For the evaluation of the non-urine specimen types, the specimens were considered non-infected.

23

AC2 Assay on PANTHER

Assay Results						Symptom Status
GC Infected Status	AC2 DTS		AGC DTS		AC2
PANTHER
MS	Sym	Asym
	MS	MU	MS	MU
Infected	+	+	+	+	+	30	2
Infected	+	+	+	+	NA	0	1
Infected	+	-	+	-	+	1	1
Infected	NA	+	NA	+	NA	1	0
Not Infected	-	-	NA	NA	-	205	307
Not Infected	-	-	NA	NA	NA	14	9

Table 14. GC Infected Status for Performance Evaluation in Male Urethral Swab Specimens

AC2 = APTIMA COMBO 2, AGC = APTIMA GC Assay, Asym = asymplomatic, DTS = DTS Systems, MS = male urechral swab, MU = male urine, NA = result not available, PANTHER = PANTHER System, Sym = symptomatic, TIGRIS = TIGRIS DTS System

Table 15. GC Infected Status for Performance Evaluation in Female Vaginal Swab, PreservCyt Solution Liquid Pap, and Endocervical Swab Specimens

| | AC2 TIGRIS | | AGC
TIGRIS | | | AC2
PANTHER | | | Symptom Status |
|-----------------------|------------|----|---------------|----|-------------|----------------|----|-----|----------------|
| GC Infected
Status | PCyt | FU | PCyt | FU | CVS/
PVS | PCyt | FS | Sym | Asym |
| Infected | + | + | + | + | + | + | + | 22 | 10 |
| Infected | + | + | + | + | + | + | NA | 1 | 0 |
| Infected | + | + | + | - | + | + | + | 1 | 0 |
| Infected | + | + | + | = | + | + | + | 0 | 1 |
| Infected | + | - | + | - | + | + | + | 3 | 3 |
| Infected | + | - | + | - | + | + | + | 0 | 1 |
| Infected | + | NA | + | NA | + | + | + | 0 | 1 |
| Not Infected | + | NA | - | - | + | = | - | 0 | 1 |
| Not Infected | - | - | NA | NA | + | - | + | 0 | 1 |
| Not Infected | - | - | NA | NA | + | - | - | 3 | 0 |
| Not Infected | - | - | NA | NA | + | - | NA | 1 | 0 |
| Not Infected | - | - | NA | NA | - | - | + | 1 | 0 |
| Not Infected | - | - | NA | NA | - | - | - | 736 | 429 |
| Not Infected | - | - | NA | NA | - | - | = | 1 | 0 |
| Not Infected | - | - | NA | NA | - | - | NA | 32 | 9 |
| Not Infected | - | - | NA | NA | - | NA | - | 1 | 0 |
| Not Infected | - | - | NA | NA | NA | - | - | 18 | 6 |
| Not Infected | - | - | NA | NA | NA | - | NA | 10 | 3 |

AC2 = APTIMA COMBO 2, ACT = APTIMA CT Assay, Asym = asympiomatic, CVS = clinician-collected vaginal swab, FS = female cndocervical swab, FU = female urine, NA = result not available, PANTHER System, PCy = PreservCy Solution liquid Pap, PVS = patient-collected vaginal swab, Sym = symptomatic, TIGRIS = TIGRIS DTS System.

The equal symbol (=) represents an equivocal result on repeat testing.

24

RLU Distribution of APTIMA COMBO 2 Controls

The distribution of the RLU values for the APTIMA COMBO 2 controls is presented in Table 16 from all valid PANTHER System runs performed during the clinical performance study.

| Control | Statistic | Total RLU
(×1000) |
|-----------------------------------------------|-----------|----------------------|
| Positive Control, CT/
Negative Control, GC | N | 66 |
| | Maximum | 1335 |
| | Median | 1081.5 |
| | Minimum | 624 |
| | CV% | 11.2 |
| Positive Control, GC/
Negative Control, CT | N | 66 |
| | Maximum | 1241 |
| | Median | 1172.0 |
| | Minimum | 1063 |
| | CV% | 3.2 |

Table 16. RLU Distribution of APTIMA COMBO 2 Controls

Reproducibility Study

APTIMA Combo 2 Assay reproducibility was evaluated at two external US laboratories and at Gen-Probe using the PANTHER System. Testing was performed using one lot of assay reagents and a total of six operators (two at each site). At each site, testing was performed over at least 10 days.

Reproducibility panel members were created using negative specimens processed with the APTIMA Specimen Collection Kit. The positive panel members were created by spiking with CT and/or GC organisms to result in panel members with expected targeted concentrations.

Table 17 shows the CT and GC concentrations for each panel member. Table 17 presents, for each panel member, RLU data in terms of mean, standard deviation (SD), and coefficient of variation (CV) between sites, between operators, between runs, within runs, and overall. Percent agreement with expected results is also shown. Samples with valid results were included in the analyses.

25

. AC2 Assay on PANTHER

Table 17. Reproducibility Data

.

·

. .

:

.

·

.

| Target
Concentration | | | | | Between
Sites | | Between
Operators | | Between
Days | | Between
Runs | | Within
Runs | | Total | |
|-------------------------|----------------|----------|--------------|------------------------|------------------|-----------|----------------------|-----------|-----------------|-----------|-----------------|-----------|----------------|-----------|---------------|-----------|
| CT
(IFU/mL) | GC
(CFU/mL) | Agreed/N | Agrmt
(%) | Mean
RLU
(x1000) | SD
(x1000) | CV
(%) | SD
(x1000) | CV
(%) | SD
(x1000) | CV
(%) | SD
(x1000) | CV
(%) | SD
(x1000) | CV
(%) | SD
(x1000) | CV
(%) |
| 0 | 0 | 178/180 | 98.9 | 6.2 | 1.2 | 19.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 8.2 | 131.7 | 8.3 | 133.0 |
| 0.25 | 0 | 180/180 | 100 | 1202.1 | 92.4 | 7.7 | 0.0 | 0.0 | 0.0 | 0.0 | 62.9 | 5.2 | 50.3 | 4.2 | 122.6 | 10.2 |
| 2.5 | 0 | 178/178 | 100 | 1184.5 | 90.9 | 7.7 | 0.0 | 0.0 | 0.0 | 0.0 | 53.8 | 4.5 | 34.6 | 2.9 | 111.1 | 9.4 |
| 25 | 0 | 180/180 | 100 | 1265.4 | 97.4 | 7.7 | 18.9 | 1.5 | 0.0 | 0.0 | 62.4 | 4.9 | 35.1 | 2.8 | 122.4 | 9.7 |
| 1000 | 0 | 180/180 | 100 | 1278.5 | 101.9 | 8.0 | 15.7 | 1.2 | 20.6 | 1.6 | 61.4 | 4.8 | 31.8 | 2.5 | 125.9 | 9.8 |
| 0 | 0.25 | 177/179 | 98.9 | 421.9 | 40.3 | 9.5 | 21.9 | 5.2 | 27.6 | 6.5 | 35.3 | 8.4 | 72.7 | 17.2 | 96.9 | 23.0 |
| 0 | 12.5 | 179/180 | 99.4 | 1141.8 | 11.9 | 1.0 | 0.0 | 0.0 | 44.4 | 3.9 | 37.3 | 3.3 | 75.8 | 6.6 | 96.2 | 8.4 |
| 0 | 125 | 180/180 | 100 | 1223.7 | 31.4 | 2.6 | 13.0 | 1.1 | 11.1 | 0.9 | 19.8 | 1.6 | 34.3 | 2.8 | 53.4 | 4.4 |
| 0 | 1250 | 180/180 | 100 | 1262.9 | 16.7 | 1.3 | 9.4 | 0.7 | 21.0 | 1.7 | 14.0 | 1.1 | 30.6 | 2.4 | 44.1 | 3.5 |
| 0 | 2500 | 180/180 | 100 | 1308.7 | 20.7 | 1.6 | 13.4 | 1.0 | 0.0 | 0.0 | 21.7 | 1.7 | 25.3 | 1.9 | 41.4 | 3.2 |
| 2.5 | 125 | 180/180 | 100 | 2467.6 | 71.9 | 2.9 | 31.5 | 1.3 | 21.7 | 0.9 | 64.8 | 2.6 | 44.4 | 1.8 | 113.1 | 4.6 |
| 2.5 | 2500 | 180/180 | 100 | 2453.3 | 76.2 | 3.1 | 30.9 | 1.3 | 0.0 | 0.0 | 62.5 | 2.5 | 51.6 | 2.1 | 115.4 | 4.7 |
| 1000 | 125 | 179/179 | 100 | 2503.8 | 74.0 | 3.0 | 38.5 | 1.5 | 0.0 | 0.0 | 59.1 | 2.4 | 39.1 | 1.6 | 109.4 | 4.4 |
| 1000 | 2500 | 180/180 | 100 | 2357.1 | 79.1 | 3.4 | 0.0 | 0.0 | 0.0 | 0.0 | 74.2 | 3.1 | 55.2 | 2.3 | 121.7 | 5.2 |

Agrm = agreement, CV = coefficient of variation, N = number of samples, RLU = relative light unit, SD = standard deviation.

: :

t

.

. .

None: Variability from some factors may be numerically negative, which can occur if the variability doe to those factors is very small. When this occuss, the variability as measured with standard deviation and %CV is set to 0. ·

.

26

DEPARTMENT OF HEALTH & HUMAN SERVICES

Image /page/26/Picture/1 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" around the perimeter. Inside the circle is a stylized image of what appears to be an abstract caduceus, a symbol often associated with healthcare.

Food and Drug Administration

10903 New Hampshire Avenue Silver Spring, MD 20993

MAY - 3 2012

0

Gen-Probe, Inc. Jody Fleming Regulatory Affairs Specialist 10210 Genetic Center Drive San Diego, CA 92121

Re: K111409

13 Trade/Device Name: APTIMA COMBO 2 Assay on the (PANTHER System) Regulation Number: 21 CFR 866.3120 Regulation Name: DNA Reagents, Neisseria Regulatory Class: II Product Code: LSL, MKZ, NSU Dated: April 20, 2012 Received: April 23, 2012

Dear Ms. Fleming:

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 class II (Special Controls), 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); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); and good manufacturing pactice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820). This letter

27

Page 2 - Jody Fleming

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 advice for your device on our labeling regulation (21 CFR Parts 801 and 809), please contact the Office of In Vitro Diagnostic Device Evaluation and Safety 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,

Jary A. Harper

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

Enclosure

28

Gen-Probe Incorporated

Indications for Use Form

510(k) Number (if known): K111409

Device Name: APTIMA COMBO 2® Assay

Indications For Use:

The PANTHER System is intended to fully automate amplified nucleic acid test (NAT) diagnostic assays currently developed by Gen-Probe Incorporated (San Diego, CA).

. .

29

Intended Use -- APTIMA COMBO 2 Assay

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) to aid in the diagnosis of chlamydial and/or gonococcal urogenital disease using the PANTHER System as specified.

On the PANTHER System, the assay may be used to test the following specimens from symplomatic and asymptomatic individuals: clinician-collected endocervical, vaginal and male urethral swab specimens, clinician-collected gynecological specimens collected in the PreservCyt Solution, and patient-collected vaginal swab specimens.

'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.

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)

Tauain Teldble

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

510(k) K111409

Page 2 of 2

1.2 510K Indication For Use Form_Final Final with K Confidential Original 510(k) Submission