← Product Code [LSL](/submissions/MI/subpart-d%E2%80%94serological-reagents/LSL) · K111409

# APTIMA COMBO 2 ASSAY (K111409)

_Gen-Probe, Inc. · LSL · May 3, 2012 · Microbiology · SESE_

**Canonical URL:** https://fda.innolitics.com/submissions/MI/subpart-d%E2%80%94serological-reagents/LSL/K111409

## Device Facts

- **Applicant:** Gen-Probe, Inc.
- **Product Code:** [LSL](/submissions/MI/subpart-d%E2%80%94serological-reagents/LSL.md)
- **Decision Date:** May 3, 2012
- **Decision:** SESE
- **Submission Type:** Traditional
- **Regulation:** 21 CFR 866.3390
- **Device Class:** Class 2
- **Review Panel:** Microbiology
- **Attributes:** Pediatric

## 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.

## Device Story

The APTIMA Combo 2 Assay is an in vitro diagnostic test for Chlamydia trachomatis and Neisseria gonorrhoeae. It uses target capture, transcription-mediated amplification (TMA), and hybridization protection assay (HPA) technologies. Specimens are collected in transport tubes that stabilize rRNA. In the laboratory, the PANTHER System automates the process: target capture using magnetic microparticles isolates rRNA; TMA amplifies specific 23S rRNA (CT) and 16S rRNA (GC) regions; HPA uses chemiluminescent DNA probes to detect amplicons. The system measures light emission as Relative Light Units (RLU). Kinetic profiles (rapid 'flasher' for CT, slower 'glower' for GC) allow differentiation. Results are determined by RLU cut-offs and kinetic curve types. The device aids clinicians in diagnosing urogenital infections. It benefits patients by providing accurate, automated detection of these pathogens from various specimen types.

## Clinical Evidence

Prospective, multicenter clinical study (n=580 males, n=1332 females) across 7 US sites. Performance compared to an infected status algorithm (two reference NAATs). For CT detection, sensitivity ranged from 97.2% to 100% and specificity from 98.5% to 100% across specimen types. For GC detection, sensitivity ranged from 97.7% to 100% and specificity from 99.6% to 100%. Analytical studies included sensitivity (LOD), precision, and carryover (0% carryover rate).

## Technological Characteristics

Uses target capture, transcription-mediated amplification (TMA), and hybridization protection assay (HPA). Detection via chemiluminescent acridinium ester-labeled DNA probes. Automated platform (PANTHER System). Qualitative assay. No changes to assay reagents from predicate.

## Regulatory 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

- TIGRIS® DTS® GEN-PROBE® APTIMA COMBO 2® Assay ([K060652](/device/K060652.md))

## Submission Summary (Full Text)

> This content was OCRed from public FDA records by [Innolitics](https://innolitics.com). If you use, quote, summarize, crawl, or train on this content, cite Innolitics at https://innolitics.com.
>
> Innolitics is a medical-device software consultancy. We help companies design, build, and clear FDA-regulated software and AI/ML devices, including [a 510(k)](https://innolitics.com/services/510ks/), [a De Novo](https://innolitics.com/services/regulatory/), [a SaMD](https://innolitics.com/services/end-to-end-samd/), [an AI/ML medical device](https://innolitics.com/services/medical-imaging-ai-development/), or [an FDA regulatory strategy](https://innolitics.com/services/regulatory/).

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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY

A. 510(k) Number:
K111409

B. Purpose for Submission:
New device clearance

C. Measurand:
Chlamydia trachomatis and Neisseria gonorrhoeae ribosomal RNA

D. Type of Test:
Qualitative in vitro diagnostic assay that utilizes Transcription-Mediated Amplification (TMA), and Hybridization Protection Assay (HPA) technologies.

E. Applicant:
Gen-Probe, Inc.

F. Proprietary and Established Names:
APTIMA COMBO 2® Assay (PANTHER® System)

G. Regulatory Information:
1. Regulation section:
866.3120
866.3390
2. Classification:
Class II
3. Product code:
MKZ
LSL
NSU

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4. Panel:

Microbiology

H. Intended Use:

1. Intended use(s):

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.¹

¹ 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.

2. Indication(s) for use:

Same as the Intended Use

3. Special conditions for use statement(s):

For Prescription Use Only

4. Special instrument requirements:

Gen-Probe PANTHER System

I. 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 target 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

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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 reaction 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.

J. Substantial Equivalence Information:

1. Predicate device name(s):
TIGRIS® DTS® GEN-PROBE® APTIMA COMBO 2® Assay

2. Predicate 510(k) number(s):
K060652

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3. Comparison with predicate:

|  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 |  | The same except using the Panther instead of Tigris system and no urine samples are cleared for use with this system.  |
|  Specimen Types | Female specimens: Urine Vaginal swab Endocervical swab ThinPrep in PreservCyt solution Male Specimens: Urine 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  |
|  Specimen Transport/Storage | 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. 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 30days. If longer storage is desired, freeze at -20°C to -70°C for up to 365 days. | Same  |
|  Type of Assay | Nucleic Acid Amplification Test | Same  |
|  Technology* | Target Capture (TC), Transcription-Mediated Amplification (TMA), Hybridization Protection Assay (HPA) | Same  |

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|  Detection Format | HPA which provides relative light units (RLUs) that are assessed against an established assay cutoff | Same  |
| --- | --- | --- |

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

NA

L. Test Principle:

See I. Device Description

M. Performance Characteristics (if/when applicable):

Analytical performance of the APTIMA COMBO 2 assay on the PANTHER System

1. 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).

2. 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.

The 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 the table below.

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Table 1. PANTHER System Within Laboratory Precision Data

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

Agrmt = agreement, CV = coefficient of variation, N = number of samples, PCyt = PreservCyt Solution liquid Pap, RLU = relative light unit, SD = standard deviation, STM = swab transport medium.
Note: Variability from some factors may be numerically negative, which can occur if the variability due to those factors is very small. When this occurs, the variability as measured with standard deviation and %CV is set to 0.

# 3. 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  $2.5 \times 10^{5} \mathrm{CFU/mL}$ . Testing was carried out using 5 runs on each 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  $0 - 0.1\%$ .

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## 4. Analytical Specificity

The APTIMA COMBO 2 Assay formulation for the PANTHER System has not changed from that used with DTS Systems or the TIGRIS DTS System. The analytical specificity study was conducted on DTS Systems. A total of 154 culture isolates were evaluated using the APTIMA COMBO 2 Assay. These isolates included 86 organisms that may be isolated from the urogenital tract and 68 additional organisms that represent a phylogenetic cross-section of organisms. The tested organisms included bacteria, fungi, yeast, parasites, and viruses. All organisms except C. psittaci, C. pneumoniae, and the viruses were tested at 1.0 x 106 cells/assay in STM. The Chlamydia and Neisseria organisms were tested in PreservCyt solution medium. C. psittaci and C. pneumoniae were tested at 1.0 x 105 IFU/assay. The viruses were tested as follows: (a) herpes simplex viruses I and II: 2.5 x 104 TCID50/assay, (b) human papilloma virus 16: 2.9 x 106 DNA copies/assay and (c) cytomegalovirus: 4.8 x 105 infected cell culture cells/assay. Only CT and GC samples produced positive results in the APTIMA COMBO 2 Assay.

## 5. Interfering Substances

The APTIMA COMBO 2 Assay formulation for the PANTHER System has not changed from that used with DTS Systems or the TIGRIS DTS System. Blood interference was evaluated on the PANTHER System and the results of this testing indicated that blood does not interfere with APTIMA COMBO 2 Assay performance.

APTIMA COMBO 2 Assay performance in the presence of potentially interfering substances was tested on DTS Systems. The following interfering substances were individually spiked into Swab and PreservCyt Solution liquid Pap specimens: 10% blood, contraceptive jelly, spermicide, moisturizer, hemorrhoidal anesthetic, body oil, powder, anti-fungal cream, vaginal lubricants, feminine spray and leukocytes (1.0 x 106 cells/mL). All were tested for potential assay interference in the absence and presence of CT and GC at the estimated rRNA equivalent of 1.0 CT IFU/assay (5 fg/assay) and 50 GC cells/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. No inhibitors of amplification were observed in the APTIMA COMBO 2 Assay.

## Clinical performance of the APTIMA COMBO 2 assay on the PANTHER System

### 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.

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

|  Site | CT+/GC- Prevalence % (# positive/# tested with valid results)  |   |   |   |
| --- | --- | --- | --- | --- |
|   |  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

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

|  Site | CT-/GC+ Prevalence % (# positive/# tested with valid results)  |   |   |   |
| --- | --- | --- | --- | --- |
|   |  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)  |

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 | CT+/GC+ Prevalence % (# positive/# tested with valid results)  |   |   |   |
| --- | --- | --- | --- | --- |
|   |  Male Urethral Swab | Clinician-Collected/Patient-collected Vaginal Swab | PreservCyt | Endocervical Swab  |

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# 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).

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

|  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  |

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|  15 | 96.1 | 99.5 | 99.1 | 100  |
| --- | --- | --- | --- | --- |
|  20 | 97.2 | 99.3 | 99.3 | 100  |
|  25 | 97.9 | 99.1 | 99.5 | 100  |

# 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  $< 18$  years of age, 72 were 18 to 20 years of age, 201 were 21 to 25 years of age, and 307 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.

{10}

# 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.

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

|  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 /PV S | 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)  |
|  PCy t | 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)  |

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 = true negative, TP = true positive.
Score CI
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.

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, as shown in Table 7.

Table 7. Performance Characteristics of the APTIMA COMBO 2 Assay for CT 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 | 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)  |

{11}

Asym = asymptomatic, 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, Sx = symptom, Sym = symptomatic, TN = true negative, TP = true positive.

Score CI
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 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.

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) |
| 1 | 225 | 27 | 0 | 198 | 0 | 12.0 | 100 (87.5-100) | 100 (98.1-100) | 100 (88.1-100) | 100 (98.3-100) |

{12}

|  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)  |

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 = true negative, TP = true positive.
Score CI
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 8. Performance Characteristics of the APTIMA COMBO 2 Assay for CT Detection by Clinical Site (continued)

|  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)  |

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 = true negative, TP = true positive.
Score CI
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.

{13}

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.

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

|  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)  |

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 = true negative, TP = true positive.
Score CI
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 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)  |
|   |  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)  |
|   |  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)  |
|   |  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)  |
|   |  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)  |

Asym = asymptomatic, 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

{14}

swab, Spec = specimen, Sx = symptom, Sym = symptomatic, TN = true negative, TP = true positive.

Score CI
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.

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

|  Spec Type MS | Site | n | TP | FP | TN | FN | Pre v % | Sensitivity % (95% CI)1 | Specificity % (95% CI)1 | PPV % (95% CI)2 | NPV% (95% CI)2  |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|  CS/PVS | 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)  |
|  PCyt | 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)  |

{15}

|   | 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)  |
|  FS | 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 | 231 | 13 | 0 | 218 | 0 | 5.6 | 100(77.2-100) | 100(98.3-100) | 100(78.0-100) | 100(98.5-100)  |
|   |  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 | 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 | 22 | 0 | 0 | 22 | 0 | 0.0 | NC | 100(85.1-100) | NC | 100(NC)  |
|   |  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 | 152 | 5 | 0 | 147 | 0 | 3.3 | 100(56.6-100) | 100(97.5-100) | 100(57.9-100) | 100(98.3-100)  |

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 = true negative, TP = true positive.
Score CI
${}^{2}$  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.

The frequency of test outcomes from reference NAAT and investigational PANTHER System testing is summarized in Table 12 and Table 13 for CT and in Tables 16 and Table 15 for GC.

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

|  CT Infected Status | Assay Results |   |   |   |   |   | Symptom Status  |   |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
|   |  AC2 DTS |   | ACT TIGRIS |   | AC2 PANTHER  |   |   |   |
|   |  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 |   |

{16}

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

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

|  CT Infected Status | Assay Results |   |   |   |   |   |   |   |   |   |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|   |  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 | + | + | 0 | 1  |   |
|  Infected | + | NA | + | NA | + | + | + | 0 | 1  |   |
|  Infected | + | NA | + | NA | - | + | - | 0 | 1  |   |
|  Infected | - | + | - | + | + | - | + | 1 | 0  |   |
|  Infected | - | + | - | + | + | - | - | 2 | 0  |   |
|  Infected | - | + | - | + | - | - | - | 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 | - | - | - | - | - | - | 0 | 1  |   |
|  Not Infected | NA | - | - | - | - | - | NA | 1 | 0  |   |
|  Not Infected | NA | - | - | - | NA | - | + | 1 | 0  |   |

{17}

AC2 = APTIMA COMBO 2, ACT = APTIMA CT Assay, Asym = asymptomatic, CVS = clinician-collected vaginal swab, FS = female endocervical swab, FU = female urine, NA = result not available, PANTHER = PANTHER System, PCyt = PreservCyt Solution liquid Pap, PVS = patient-collected vaginal swab, Sym = symptomatic, TIGRIS = TIGRIS DTS System

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

|  GC Infected Status | Assay Results |   |   |   |   | Symptom Status  |   |
| --- | --- | --- | --- | --- | --- | --- | --- |
|   |  AC2 DTS |   | AGC DTS |   | AC2 PANTHER | Sym | Asym  |
|   |  MS | MU | MS | MU | MS  |   |   |
|  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  |

AC2 = APTIMA COMBO 2, AGC = APTIMA GC Assay, Asym = asymptomatic, DTS = DTS Systems, MS = male urethral 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

|  GC Infected Status | Assay Results |   |   |   |   |   |   |   | Symptom Status  |   |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|   |  AC2 TIGRIS |   | AGC TIGRIS |   | AC2 PANTHER  |   |   |   |   |   |
|   |  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 |   |

{18}

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

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

## 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.

Table 16. RLU Distribution of APTIMA COMBO 2 Controls

|  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  |

## 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 days, between runs, within runs, and overall. Percent agreement with expected results is also shown. Samples with valid results were included in the analyses.

19

{19}

Table 17. Reproducibility Data

|  Target Concentration |   | Agreed/N | Agrmt (%) | Mean RLU (x1000) | Between Sites |   | Between Operators |   | Between Days |   | Between Runs |   | Within Runs |   | Total  |   |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|  CT (IFU/mL) | GC (CFU/mL) |   |   |   | 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  |

Agrmt = agreement, CV = coefficient of variation, N = number of samples, RLU = relative light unit, SD = standard deviation.
Note: Variability from some factors may be numerically negative, which can occur if the variability due to those factors is very small.
When this occurs, the variability as measured with standard deviation and  $\% \mathrm{CV}$  is set to 0.

# N. Instrument Name:

PANTHER System

# O. System Descriptions:

1. Modes of Operation:

Batch, Random access

{20}

2. Software:

FDA has reviewed applicant’s Hazard Analysis and software development processes for this line of product types:

Yes ☐ X ☐ or No ☐

3. Specimen Identification:

By hand-held barcode reader and positional checks

4. Specimen Sampling and Handling:

Fully automated

5. Calibration:

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

6. Quality Control:

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

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

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

Q. Proposed Labeling:

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

R. Conclusion:

The submitted information in this premarket notification is complete and supports a substantial equivalence decision.

---

**Source:** [https://fda.innolitics.com/submissions/MI/subpart-d%E2%80%94serological-reagents/LSL/K111409](https://fda.innolitics.com/submissions/MI/subpart-d%E2%80%94serological-reagents/LSL/K111409)

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