Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The description focuses on standard PCR technology and automated sample processing, with no mention of AI or ML algorithms for data analysis or interpretation.

Therapeutic

No
This device is an in vitro diagnostic test intended as an aid in the diagnosis of chlamydial and gonococcal disease, not for therapeutic purposes.

Diagnostic

Yes

The first sentence of the "Intended Use / Indications for Use" section explicitly states: "The cobas® CT/NG on the cobas® 6800/8800 system is an automated, qualitative in vitro nucleic acid diagnostic test..." and further clarifies it is "intended as an aid in the diagnosis of chlamydial and gonococcal disease".

SaMD (Software as a Medical Device)

No

The device description explicitly states that the cobas® CT/NG is a test performed on the cobas® 6800 System and cobas® 8800 System, which are described as consisting of hardware components (sample supply module, transfer module, processing module, analytic module). While the system includes software for data management, the device itself is a combination of hardware and software for performing a diagnostic test.

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 cobas® CT/NG on the cobas® 6800/8800 system is an "automated, qualitative in vitro nucleic acid diagnostic test".
Device Description: The description details how the test works by detecting DNA from Chlamydia trachomatis and Neisseria gonorrhoeae in various human specimens (urine, vaginal swabs, endocervical swabs, cervical specimens). This process of analyzing biological samples outside of the body to diagnose a condition is the core definition of an in vitro diagnostic.
Clinical Performance: The document provides extensive data on the clinical performance of the test, including sensitivity, specificity, PPV, and NPV, which are standard metrics for evaluating the accuracy of diagnostic tests.
Predicate Device: The mention of a "Predicate Device(s)" with a K number (K163184) indicates that this device is being compared to a previously cleared medical device, which is a common process for IVDs seeking regulatory approval.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The cobas® CT/NG on the cobas® 6800/8800 system is an automated, qualitative in vitro nucleic acid diagnostic test, that utilizes real-time polymerase chain reaction (PCR), for the direct detection of Chlamydia trachomatis (CT) and/or Neisseria gonorthoeae (NG) DNA in male and female urine, clinician-instructed self-collected vaginal swab specimens (collected in a clinical setting), clinician-collected vaginal swab specimens, and endocervical swab specimens, all collected in cobas® PCR Media (Roche Molecular Systems, Inc.), and cervical specimens collected in PreservCyt® solution. This test is intended as an aid in the diagnosis of chlamydial and gonococcal disease in both symptomatic and asymptomatic individuals.

Ancillary Collection Kits:
The cobas® PCR Media Dual Swab Sample Kit is used to collect and transport endocervical and vaginal swab specimens. The cobas® PCR Media serves as a nucleic acid stabilizing transport and storage medium for gynecological specimens.
Note: This kit has been validated for use with the following tests:
• cobas® CT/NG v2.0 Test
• cobas® CT/NG for use on cobas® 6800/8800 Systems
The cobas® PCR Media Uni Swab Sample Kit is used to collect and transport human specimens. The cobas® PCR Media serves as a nucleic acid stabilizing transport and storage medium for human specimens.
Note: This kit has been validated for use with the following tests:
• cobas® CT/NG v2.0 Test
• cobas® CT/NG for use on cobas® 6800/8800 Systems
• cobas® Cdiff Test for use on the cobas® 4800 System
The cobas® PCR Urine Sample Kit is used to collect and transport urine specimens. The cobas® PCR Media serves as a nucleic acid stabilizing transport and storage medium for urine specimens. Use this cobas® CT/NG on cobas® 6800/8800 Systems or the cobas® CT/NG v2.0 Test.

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

LSL, MKZ, OOI

Device Description

cobas® CT/NG is a new qualitative test performed on the cobas® 6800 System and cobas® 8800 System. cobas® CT/NG enables the detection of CT/NG DNA in endocervical, vaginal, urine and cervical specimens of infected female patients and urine specimens in infected male patients. Target-specific primers and two probes are used to detect but not discriminate between the CT cryptic plasmid and the ompA gene. Additionally, target-specific primers and two probes are used to detect but not discriminate between two conserved sequences in the NG DR-9 region. The DNA Internal Control, used to monitor the entire sample preparation and PCR amplification process, is introduced into each specimen during sample processing. In addition, the test utilizes a low titer positive and a negative control.

Principles of the procedure
cobas® CT/NG is based on fully automated sample preparation (nucleic acid extraction and purification) followed by PCR amplification and detection. The cobas® 6800/8800 Systems consist of the sample supply module, the transfer module, the processing module, and the analytic module. Automated data management is performed by the cobas® 6800/8800 software which assigns test results for all tests as positive, negative or invalid. Results can be reviewed directly on the system screen, exported, or printed as a report.

Nucleic acid from patient samples, external controls and added internal control DNA (DNA-IC) molecules is simultaneously extracted. In summary, bacterial nucleic acid is released by addition of proteinase and lysis reagent to the sample. The released nucleic acid binds to the silica surface of the added magnetic glass particles. Unbound substances and impurities, such as denatured protein, cellular debris and potential PCR inhibitors are removed with subsequent wash steps and purified nucleic acid is eluted from the magnetic glass particles with elution buffer at elevated temperature.

Selective amplification of target nucleic acid from the sample is achieved by the use of targetspecific forward and reverse primers which are selected from highly conserved plasmid and genomic regions of CT and NG. A region on the CT cryptic plasmid and the ompA gene (dual target) and two conserved sequences of the NG DR-9 region are amplified by cobas® CT/NG.

Selective amplification of DNA-IC is achieved by the use of sequence-specific forward and reverse primers which are selected to have no homology with either the CT or NG target regions. A thermostable DNA polymerase enzyme is used for PCR amplification. The target and DNA-IC sequences are amplified simultaneously utilizing a universal PCR amplification profile with predefined temperature steps and number of cycles. The master mix includes deoxyuridine triphosphate (dUTP), instead of deoxythimidine triphosphate (dTTP), which is incorporated into the newly synthesized DNA (amplicon). Any contaminating amplicon from previous PCR runs are eliminated by the AmpErase enzyme, which is included in the PCR master mix, during the first thermal cycling step. However, newly formed amplicons are not eliminated since the AmpErase enzyme is inactivated once exposed to temperatures above 55℃.

The cobas® CT/NG master mix contains two detection probes specific for the CT target sequences, two detection probes specific for the NG target sequences and one for the DNA-IC. The probes are labeled with target specific fluorescent reporter dyes allowing simultaneous detection of CT targets, NG targets and DNA-IC in three different target channels. When not bound to the target sequence, the fluorescent signal of the intact probes is suppressed by a quencher dye. During the PCR amplification step, hybridization of the probes to the specific single-stranded DNA template results in cleavage of the 5' to 3' exonuclease activity of the DNA polymerase resulting in separation of the reporter and quencher dyes and the generation of a fluorescent signal. With each PCR cycle, increasing amounts of cleaved probes are generated and the cumulative signal of the reporter dye increases concomitantly. Real-time detection and discrimination of PCR products is accomplished by measuring the fluorescence of the released reporter dyes for the CT and NG targets and DNA-IC, respectively.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

male and female urine, clinician-instructed self-collected vaginal swab specimens (collected in a clinical setting), clinician-collected vaginal swab specimens, and endocervical swab specimens, all collected in cobas® PCR Media (Roche Molecular Systems, Inc.), and cervical specimens collected in PreservCyt® solution.

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 total of 5.197 subjects were prospectively enrolled, of which 5.105 were eligible for inclusion. Of the 5,105 eligible subjects contributing prospective specimens, 5,053 (99.0%) (3,860 females and 1,193 males) were evaluable and were included in the data analyses. A total of 52 subjects (1.0%) were classified as non-evaluable and excluded from all statistical analyses. There were a total of 371 archived prospectively collected female urogenital samples (urine, cervical specimens in PreservCyt, and endocervical swabs) tested in this clinical study from 295 female subjects. Among the 17.169 samples tested in this study, 19 samples exhibited invalid results on the first run (invalid rate of 0.11% (95%CI: 0.07% ; 0.17%)). Upon repeat testing, 3 samples exhibited valid results.

Female Study Design:
Prospectively enrolled female subjects provided the following urogenital specimens: first-void urine, 3 vaginal swabs, 1 endocervical swab in cobas® PCR Media, and 1 cervical sample in PreservCyt® Solution. If the female was in the clinician-collected vaginal swab arm of the study, 2 of the vaginal swabs were placed in the respective manufacturers' collection device and 1 in cobas® PCR Media. If the female subject was in the self-collected vaginal swab self-collection arm of the study, then 1 vaginal swab was self-collected first and placed into cobas® PCR Media and then followed by the 2 clinician-collected vaginal swabs and placed in the 2 respective manufacturers' collection devices.

Male Study Design:
Prospectively enrolled male subjects provided a urine specimen that was aliquoted into the respective manufacturers' collection device and cobas® PCR Media.

Symptom Classification:
Subjects were classified as symptomatic if they self-reported symptoms indicative of a CT or NG infection as listed below:

Dysuria (pain during urination) .
Coital pain, difficulty or bleeding .
Pelvic pain .
Abnormal vaginal discharge .
Pelvic, uterine or ovarian pain .
Urethral discharge .
Testicular pain .
Scrotal pain or swelling .
Prospectively enrolled subjects were classified as asymptomatic if they did not report any of the above symptoms.

PIS Determination:
The clinical performance of cobas® CT/NG was evaluated by comparing the results from collected specimen types to a pre-specified PIS (Patient Infected Status) algorithm as determined by the combined results from 2 commercially available NAATs for females and 3 commercially available NAATs for males. All tests were run according to the respective manufacturers' Instructions For Use.
For NG, archived prospectively collected female urine, cervical specimens in PreservCyt®, and endocervical swabs were obtained from the clinical study for cobas® CT/NG v2 test on the cobas® 4800 System. The PIS of these specimens were already determined from the clinical study for cobas® CT/NG v2 test on the cobas® 4800 System.

PIS Algorithm for Female Urogenital Specimens (Table 11):

NAAT1 Urine/Vaginal	NAAT2 Urine/Vaginal	Patient Infected Status (PIS)
+/+	+/+	Infected
+/+	+/- or -/+	Infected
+/- or -/+	+/+	Infected
+/-	-/+	Infected
-/+	+/- or -/+	Infected
+/-	+/-	Infected (Urine) / Non-Infected (Vaginal)
+/- or -/+	-/-	Not Infected
+/+	-/-	Not Infected
-/-	+/+	Not Infected
-/-	+/- or -/+	Not Infected
-/-	-/-	Not Infected
Note: One or more positives in each NAAT (NAAT1 and NAAT2) designates the PIS as Infected. Any other combination of results defines the PIS as Not Infected.
If one or more of the sample types are invalid, the remaining sample types with valid results from NAAT1 and NAAT2 must have concordant positive or concordant negative results to determine the PIS as Infected or Not Infected, respectively. For all other cases where one or more of the sample types are invalid, the PIS is indeterminate.

PIS Algorithm for Male Urine Specimens (Table 12):
| NAAT1 Urine | NAAT2 Urine | NAAT3 Urine | Patient Infected Status (PIS) |
|---|---|---|
| + | + | + | Infected |
| + | + | - | Infected |
| + | - | + | Infected |
| - | + | + | Infected |
| - | - | + | Not Infected |
| - | + | - | Not Infected |
| + | - | - | Not Infected |
| - | - | - | Not Infected |
Note: If at least 2 out of the 3 test results are concordant positive then the PIS can be considered as Infected or Not Infected, respectively. If one test result is invalid/missing and the other two test results are discordant then the PIS is indeterminate. If 2 or 3 test results are invalid/missing, then the PIS is indeterminate.

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

Non-Clinical Performance Evaluation

Limit of Detection (LoD): Determined by analyzing a dilution series of quantified cultures of Chlamydia trachomatis (serovars D and I) and Neisseria gonorrhoeae isolates 2948 (ATCC 19424) and 891. CT and NG cultures were diluted into a matrix of pooled negative specimens of each sample type. 70-78 replicates were tested for each level in each specimen type across 3 unique lots of reagents.
- Results (Table 2):
  - Endocervical Swab in cobas® PCR: CT Serovar D LoD = 0.3 IFU/mL; CT Serovar I LoD = 1.4 IFU/mL; NG Strain 2948 LoD = 0.4 CFU/mL; NG Strain 891 LoD = 0.08 CFU/mL.
  - Vaginal Swab in cobas® PCR: CT Serovar D LoD = 0.3 IFU/mL; CT Serovar I LoD = 1.4 IFU/mL; NG Strain 2948 LoD = 0.4 CFU/mL; NG Strain 891 LoD = 0.08 CFU/mL.
  - Urine: CT Serovar D LoD = 0.2 IFU/mL; CT Serovar I LoD = 1.3 IFU/mL; NG Strain 2948 LoD = 0.2 CFU/mL; NG Strain 891 LoD = 0.04 CFU/mL.
  - Cervical Samples in PreservCyt®: CT Serovar D LoD = 0.6 IFU/mL; CT Serovar I LoD = 2.9 IFU/mL; NG Strain 2948 LoD = 0.2 CFU/mL; NG Strain 891 LoD = 0.08 CFU/mL.
Inclusivity: Tested 13 additional CT serovars and 43 independently isolated strains of NG using one lot of reagents. Testing performed using CT and NG cultures diluted into pools of negative specimens. Twenty replicates per dilution level were tested for each strain in each specimen type.
- Results (Table 3 for CT, Table 4 for NG): All CT serovars and NG strains tested showed 100% positivity or ≥95% positivity at specified IFU/mL or CFU/mL concentrations.
Precision: In-house precision examined using a panel composed of CT and NG cultures diluted into a pool of negative endocervical swab specimen matrix (in cobas® PCR Media), a pool of negative urine matrix (plus cobas® PCR Media), and a pool of negative cervical specimen matrix (in PreservCyt® Solution). Four levels tested (high negative, low, and moderate concentrations of CT and NG), corresponding to ~0.3x, ~1x, and ~3x LoD. Tested with three lots of cobas® CT/NG reagents and two instruments for a total of 24 runs.
- Results (Table 5, 6, 7): All negative panel members tested negative. Overall CV (%) ranges from 1.62% to 4.05% for CT and from 1.17% to 3.55% for NG for positive panel members.
Analytical specificity/Cross-reactivity: Panel of 149 bacteria, fungi, and viruses tested. Organisms spiked at approximately 1 x 10^6 units/mL for bacteria and 1 x 10^5 units/mL for viruses into pools of negative vaginal swab specimens (in cobas® PCR Media), urine (stabilized in cobas® PCR Media), and cervical specimens (in PreservCyt® Solution). Tested with each potential interfering organism alone and mixed with CT and NG cultures at ~3x LoD.
- Results: None of the tested organisms interfered with the detection of CT and NG or produced false positive results.
Interference: Effects of over-the-counter or prescription products and endogenous substances present in urogenital specimens evaluated. Tested using pooled clinical specimens (vaginal swab, urine, and PreservCyt®) with spiking of potential interferents at levels expected from normal patient usage. Interferents tested in CT/NG negative specimen pools and in specimen pools with CT/NG at ~3x LoD. CT serovars D and I and NG strains 2948 and 891 used.
- Results (Table 9 for OTC/prescription, Table 10 for endogenous): Metronidazole Vaginal Gel, Replens™ Long-Lasting Vaginal Moisturizer, RepHresh™ Odor Eliminating Vaginal Gel, and RepHresh™ Clean Balance produced false negative or invalid results in at least one replicate. Interference noted with whole blood at 10% for urine and PreservCyt® specimens, and with cervical mucus at 1% in endocervical specimens.
Competitive Inhibition: Evaluated by testing samples with low and moderate concentrations of one target mixed with very high concentrations of the opposite target in vaginal swab, urine, and PreservCyt® specimens.
- Results: When NG was at high concentration, CT was detected in all specimen types at low (~1x LoD) and moderate (~3x LoD) levels. When CT was at high concentration, NG was detected in all specimen types at moderate (~3x LoD) levels, but not consistently at low levels.
Cross contamination/Carryover: Evaluated potential cross-contamination on the cobas® 6800/8800 Systems using cobas® CT/NG by alternating very high positive and negative samples.
- Results: Sample-to-sample cross-contamination rate of 0.5% (2/432), (95% CI: 0.1%-1.7%). Run-to-run cross-contamination not observed (0/282).

Clinical Performance Evaluation
Study Type: Multi-site, prospective collection study comparing cobas® CT/NG results to a Patient Infected Status (PIS) algorithm using a combination of FDA-cleared NAATs.
Sites: 9 geographically diverse sites in the US for specimen collection, 4 laboratory testing sites (3 external, 1 internal).
Sample Size: 5,053 evaluable subjects (3,860 females and 1,193 males) from prospective enrollment. 371 archived prospectively collected female urogenital samples from 295 female subjects.
Key Results (CT, Table 15):

Female Urine:
- Overall Sensitivity: 95.6% (259/271) [95% CI: 92.4%, 97.4%]
- Overall Specificity: 99.7% (3577/3588) [95% CI: 99.5%, 99.8%]
Female Clinician-collected Vaginal Swab (VS-C):
- Overall Sensitivity: 98.6% (146/148) [95% CI: 95.2%, 99.6%]
- Overall Specificity: 99.1% (1772/1788) [95% CI: 98.6%, 99.4%]
Female Self-collected Vaginal Swab (VS-S):
- Overall Sensitivity: 99.2% (119/120) [95% CI: 95.4%, 99.9%]
- Overall Specificity: 99.0% (1769/1786) [95% CI: 98.5%, 99.4%]
Female PreservCyt®:
- Overall Sensitivity: 92.5% (247/267) [95% CI: 88.7%, 95.1%]
- Overall Specificity: 99.6% (3570/3584) [95% CI: 99.3%, 99.8%]
Female Endocervical Swab (ES):
- Overall Sensitivity: 93.3% (249/267) [95% CI: 89.6%, 95.7%]
- Overall Specificity: 99.4% (3553/3576) [95% CI: 99.0%, 99.6%]
Male Urine:
- Overall Sensitivity: 100.0% (118/118) [95% CI: 96.8%, 100.0%]
- Overall Specificity: 99.7% (1071/1074) [95% CI: 99.2%, 99.9%]

Key Results (NG, Table 18 - Prospective and Archived Specimens):

Female Urine (Prospective and Archived):
- Overall Sensitivity: 94.8% (127/134) [95% CI: 89.6%, 97.4%]
- Overall Specificity: 99.9% (3917/3920) [95% CI: 99.8%, 100.0%]
Female Clinician-collected Vaginal Swab (VS-C):
- Overall Sensitivity: 100.0% (28/28) [95% CI: 87.9%, 100.0%]
- Overall Specificity: 99.7% (1903/1908) [95% CI: 99.4%, 99.9%]
Female Self-collected Vaginal Swab (VS-S):
- Overall Sensitivity: 100.0% (28/28) [95% CI: 87.9%, 100.0%]
- Overall Specificity: 99.7% (1873/1879) [95% CI: 99.3%, 99.9%]
Female PreservCyt® (Prospective and Archived):
- Overall Sensitivity: 96.6% (86/89) [95% CI: 90.6%, 98.8%]
- Overall Specificity: 99.9% (3831/3833) [95% CI: 99.8%, 100.0%]
Female Endocervical Swab (ES) (Prospective and Archived):
- Overall Sensitivity: 97.0% (97/100) [95% CI: 91.5%, 99.0%]
- Overall Specificity: 99.9% (3846/3848) [95% CI: 99.8%, 100.0%]
Male Urine Reported as Overall (Table 18 shows Male UR, but not distinguishing between prospective/archived nor explicit overall for male):
- Overall Sensitivity: 100.0% (87/87) [95% CI: 95.8%, 100.0%]
- Overall Specificity: 99.5% (1099/1105) [95% CI: 98.8%, 99.8%]

Clinical Reproducibility Study
Study Design: Performed across different sites (two external, one internal), lots, operators/batches days. Used three panels prepared from swabs and urine in cobas® PCR Media and cervical specimens in PreservCyt® Solution, each with six concentrations and three replicates per concentration (54 samples per panel).
Sample Size: Total of 3,888 tests (1,296 tests for each panel type).
Key Results:

Negative panel results: 100% negative percent agreement for both CT and NG (no false positives).
Chlamydia trachomatis results (Table 19): Total coefficient of variation among positive panel members ranged from 0.9% to 3.2%. Most variability explained by random error (within-batch).
Neisseria gonorrhoeae results (Table 23): Total coefficient of variation among positive panel members ranged from 1.0% to 3.1%. Most variability explained by random error (within-batch).
Percentage agreement (Table 27):
- For panel members at or near 1x LoD, the lower limit of the 2-sided 95% exact CI was ≥ 97.4% for CT and ≥ 95.3% for NG.
- For panel members at 3x LoD, the lower limit of the 2-sided 95% exact CI was 98.3% for both CT and NG.

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

CT Clinical Performance (Table 15):

Female Urine:
- Sensitivity: Symptomatic 96.0%, Asymptomatic 95.2%, Overall 95.6%
- Specificity: Symptomatic 99.8%, Asymptomatic 99.6%, Overall 99.7%
- PPV: Symptomatic 98.3, Asymptomatic 94.0, Overall 95.9
- NPV: Symptomatic 99.6, Asymptomatic 99.7, Overall 99.7
Female Clinician-collected Vaginal Swab (VS-C):
- Sensitivity: Symptomatic 100.0%, Asymptomatic 97.6%, Overall 98.6%
- Specificity: Symptomatic 99.2%, Asymptomatic 99.0%, Overall 99.1%
- PPV: Symptomatic 92.6, Asymptomatic 88.3, Overall 90.1
- NPV: Symptomatic 100.0, Asymptomatic 99.8, Overall 99.9
Female Self-collected Vaginal Swab (VS-S):
- Sensitivity: Symptomatic 100.0%, Asymptomatic 98.4%, Overall 99.2%
- Specificity: Symptomatic 98.8%, Asymptomatic 99.2%, Overall 99.0%
- PPV: Symptomatic 88.1, Asymptomatic 87.0, Overall 87.5
- NPV: Symptomatic 100.0, Asymptomatic 99.9, Overall 99.9
Female PreservCyt®:
- Sensitivity: Symptomatic 95.1%, Asymptomatic 90.3%, Overall 92.5%
- Specificity: Symptomatic 99.5%, Asymptomatic 99.7%, Overall 99.6%
- PPV: Symptomatic 94.3, Asymptomatic 94.9, Overall 94.6
- NPV: Symptomatic 99.5, Asymptomatic 99.4, Overall 99.4
Female Endocervical Swab (ES):
- Sensitivity: Symptomatic 95.9%, Asymptomatic 91.1%, Overall 93.3%
- Specificity: Symptomatic 99.1%, Asymptomatic 99.5%, Overall 99.4%
- PPV: Symptomatic 90.6, Asymptomatic 92.4, Overall 91.5
- NPV: Symptomatic 99.6, Asymptomatic 99.4, Overall 99.5
Male Urine:
- Sensitivity: Symptomatic 100.0%, Asymptomatic 100.0%, Overall 100.0%
- Specificity: Symptomatic 99.6%, Asymptomatic 99.8%, Overall 99.7%
- PPV: Symptomatic 98.4, Asymptomatic 96.5, Overall 97.5
- NPV: Symptomatic 100.0, Asymptomatic 100.0, Overall 100.0

NG Clinical Performance (Table 18):

Female Urine (Prospective and Archived):
- Sensitivity: Symptomatic 96.7%, Asymptomatic 93.2%, Overall 94.8%
- Specificity: Symptomatic 99.8%, Asymptomatic 100.0%, Overall 99.9%
- PPV: Symptomatic 95.2, Asymptomatic 100.0, Overall 97.7
- NPV: Symptomatic 99.9, Asymptomatic 99.8, Overall 99.8
Female Clinician-collected Vaginal Swab (VS-C):
- Sensitivity: Symptomatic 100.0%, Asymptomatic 100.0%, Overall 100.0%
- Specificity: Symptomatic 99.7%, Asymptomatic 99.8%, Overall 99.7%
- PPV: Symptomatic 84.6, Asymptomatic 85.0, Overall 84.8
- NPV: Symptomatic 100.0, Asymptomatic 100.0, Overall 100.0
Female Self-collected Vaginal Swab (VS-S):
- Sensitivity: Symptomatic 100.0%, Asymptomatic 100.0%, Overall 100.0%
- Specificity: Symptomatic 99.7%, Asymptomatic 99.7%, Overall 99.7%
- PPV: Symptomatic 87.5, Asymptomatic 77.8, Overall 82.4
- NPV: Symptomatic 100.0, Asymptomatic 100.0, Overall 100.0
Female PreservCyt® (Prospective and Archived):
- Sensitivity: Symptomatic 97.9%, Asymptomatic 95.1%, Overall 96.6%
- Specificity: Symptomatic 99.9%, Asymptomatic 100.0%, Overall 99.9%
- PPV: Symptomatic 97.9, Asymptomatic 97.5, Overall 97.7
- NPV: Symptomatic 99.9, Asymptomatic 99.9, Overall 99.9
Female Endocervical Swab (ES) (Prospective and Archived):
- Sensitivity: Symptomatic 100.0%, Asymptomatic 94.5%, Overall 97.0%
- Specificity: Symptomatic 99.9%, Asymptomatic 100.0%, Overall 99.9%
- PPV: Symptomatic 97.8, Asymptomatic 98.1, Overall 98.0
- NPV: Symptomatic 100.0, Asymptomatic 99.9, Overall 99.9
Male Urine:
- Sensitivity: Symptomatic 100.0%, Asymptomatic 100.0%, Overall 100.0%
- Specificity: Symptomatic 98.7%, Asymptomatic 99.7%, Overall 99.5%
- PPV: Symptomatic 96.5, Asymptomatic 62.5, Overall 93.5
- NPV: Symptomatic 100.0, Asymptomatic 100.0, Overall 100.0

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.

cobas® CT/NG v2.0 Test (K163184)

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.

Not Found

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

Image /page/0/Picture/0 description: The image shows the logos of the Department of Health & Human Services and the U.S. Food & Drug Administration (FDA). The Department of Health & Human Services logo is on the left, and the FDA logo is on the right. The FDA logo is a blue square with the letters "FDA" in white, followed by the words "U.S. Food & Drug Administration" in blue.

March 21, 2018

Roche Molecular Systems, Inc. Nobuko Nakajima Manager, Regulatory Affairs 4300 Hacienda Drive Pleasanton, California 94588-2722

Re: K173887

Trade/Device Name: cobas CT/NG for use on cobas 6800/8800 systems Regulation Number: 21 CFR 866.3390 Regulation Name: Neisseria spp. direct serological test reagents Regulatory Class: Class II Product Code: LSL, MKZ, OOI Dated: December 20, 2017 Received: December 21, 2017

Dear Nobuko Nakajima:

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. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR

1

Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

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.

For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely,

Tamara V. Feldblyum -S for

Uwe Scherf, Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

2

Indications for Use

510(k) Number (if known) K173887

Device Name

cobas® CT/NG for use on the cobas® 6800/8800 Systems

Indications for Use (Describe)

The cobas® CT/NG on the cobas® 6800/8800 system is an automated, qualitative in vitro nucleic acid diagnostic test, that utilizes real-time polymerase chain reaction (PCR), for the direct detection of Chlamydia trachomatis (CT) and/or Neisseria gonorthoeae (NG) DNA in male and female urine, clinician-instructed self-collected vaginal swab specimens (collected in a clinical setting), clinician-collected vaginal swab specimens, and endocervical swab specimens, all collected in cobas® PCR Media (Roche Molecular Systems, Inc.), and cervical specimens collected in PreservCyt® solution. This test is intended as an aid in the diagnosis of chlamydial and gonococcal disease in both symptomatic and asymptomatic individuals.

Ancillary Collection Kits:

The cobas® PCR Media Dual Swab Sample Kit is used to collect and transport endocervical and vaginal swab specimens. The cobas® PCR Media serves as a nucleic acid stabilizing transport and storage medium for gynecological specimens.

Note: This kit has been validated for use with the following tests:

· cobas® CT/NG v2.0 Test

· cobas® CT/NG for use on cobas® 6800/8800 Systems
The cobas® PCR Media Uni Swab Sample Kit is used to collect and transport human specimens. The cobas® PCR Media serves as a nucleic acid stabilizing transport and storage medium for human specimens.

Note: This kit has been validated for use with the following tests:

· cobas® CT/NG v2.0 Test
• cobas® CT/NG for use on cobas® 6800/8800 Systems

· cobas® Cdiff Test for use on the cobas® 4800 System

The cobas® PCR Urine Sample Kit is used to collect and transport urine specimens. The cobas® PCR Media serves as a nucleic acid stabilizing transport and storage medium for urine specimens. Use this cobas® CT/NG on cobas® 6800/8800 Systems or the cobas® CT/NG v2.0 Test.

Type of Use (Select one or both, as applicable)

X Prescription Use (Part 21 CFR 801 Subpart D)

Over-The-Counter Use (21 CFR 801 Subpart C)

CONTINUE ON A SEPARATE PAGE IF NEEDED.

3

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4

cobas® CT/NG for use on the cobas® 6800/8800 Systems 510(k) Summary

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

Submitter Name	Roche Molecular Systems, Inc.
Address	4300 Hacienda Drive
Pleasanton, CA, 94588-2722
Contact	Nobuko Nakajima
Phone: (925)730-8215
FAX: (925)225-0207
Email: nobuko.nakajima@roche.com
Date Prepared	December 4, 2017
Proprietary Name	cobas® CT/NG
for use on cobas® 6800/8800 systems
Common Name	Real-time PCR assay, in vitro nucleic acid amplification test for the quantitative
detection of Chlamydia trachomatis (CT) and/or Neisseria gonorrhoeae (NG)
Classification Name	Sec. 866.3390 Neisseria spp. direct serological test reagents
Sec. 866.3120 Chlamydia serological reagents,
Sec. 862.2570 Real Time Nucleic Acid Amplification System
Product Codes	LSL
MKZ
OOI
Predicate Devices	cobas® CT/NG v2.0 Test (K163184)
Establishment Registration	Roche Molecular Systems, Inc. (2243471)

5

1. DEVICE DESCRIPTION

cobas® CT/NG is a new qualitative test performed on the cobas® 6800 System and cobas® 8800 System. cobas® CT/NG enables the detection of CT/NG DNA in endocervical, vaginal, urine and cervical specimens of infected female patients and urine specimens in infected male patients. Target-specific primers and two probes are used to detect but not discriminate between the CT cryptic plasmid and the ompA gene. Additionally, target-specific primers and two probes are used to detect but not discriminate between two conserved sequences in the NG DR-9 region. The DNA Internal Control, used to monitor the entire sample preparation and PCR amplification process, is introduced into each specimen during sample processing. In addition, the test utilizes a low titer positive and a negative control.

Principles of the procedure 1.1.

cobas® CT/NG is based on fully automated sample preparation (nucleic acid extraction and purification) followed by PCR amplification and detection. The cobas® 6800/8800 Systems consist of the sample supply module, the transfer module, the processing module, and the analytic module. Automated data management is performed by the cobas® 6800/8800 software which assigns test results for all tests as positive, negative or invalid. Results can be reviewed directly on the system screen, exported, or printed as a report.

Nucleic acid from patient samples, external controls and added internal control DNA (DNA-IC) molecules is simultaneously extracted. In summary, bacterial nucleic acid is released by addition of proteinase and lysis reagent to the sample. The released nucleic acid binds to the silica surface of the added magnetic glass particles. Unbound substances and impurities, such as denatured protein, cellular debris and potential PCR inhibitors are removed with subsequent wash steps and purified nucleic acid is eluted from the magnetic glass particles with elution buffer at elevated temperature.

Selective amplification of target nucleic acid from the sample is achieved by the use of targetspecific forward and reverse primers which are selected from highly conserved plasmid and genomic regions of CT and NG. A region on the CT cryptic plasmid and the ompA gene (dual target) and two conserved sequences of the NG DR-9 region are amplified by cobas® CT/NG.

6

Selective amplification of DNA-IC is achieved by the use of sequence-specific forward and reverse primers which are selected to have no homology with either the CT or NG target regions. A thermostable DNA polymerase enzyme is used for PCR amplification. The target and DNA-IC sequences are amplified simultaneously utilizing a universal PCR amplification profile with predefined temperature steps and number of cycles. The master mix includes deoxyuridine triphosphate (dUTP), instead of deoxythimidine triphosphate (dTTP), which is incorporated into the newly synthesized DNA (amplicon). Any contaminating amplicon from previous PCR runs are eliminated by the AmpErase enzyme, which is included in the PCR master mix, during the first thermal cycling step. However, newly formed amplicons are not eliminated since the AmpErase enzyme is inactivated once exposed to temperatures above 55℃.

The cobas® CT/NG master mix contains two detection probes specific for the CT target sequences, two detection probes specific for the NG target sequences and one for the DNA-IC. The probes are labeled with target specific fluorescent reporter dyes allowing simultaneous detection of CT targets, NG targets and DNA-IC in three different target channels. When not bound to the target sequence, the fluorescent signal of the intact probes is suppressed by a quencher dye. During the PCR amplification step, hybridization of the probes to the specific single-stranded DNA template results in cleavage of the 5' to 3' exonuclease activity of the DNA polymerase resulting in separation of the reporter and quencher dyes and the generation of a fluorescent signal. With each PCR cycle, increasing amounts of cleaved probes are generated and the cumulative signal of the reporter dye increases concomitantly. Real-time detection and discrimination of PCR products is accomplished by measuring the fluorescence of the released reporter dyes for the CT and NG targets and DNA-IC, respectively.

Figure 1: cobas® CT/NG on the cobas® 6800/8800 system

[IVD]
cobas CT/NG Positive Control Kit	cobas CT/NG	cobas Buffer Negative Control Kit
Positive control kit for use on the cobas 6800/8800 Systems	Qualitative nucleic acid test for use on the cobas 6800/8800 Systems	Buffer negative control kit for use on the cobas 6800/8800 Systems

cobas	cobas	cobas

7

2. INTENDED USE

The cobas® CT/NG on the cobas® 6800/8800 system is an automated, qualitative in vitro nucleic acid diagnostic test, that utilizes real-time polymerase chain reaction (PCR), for the direct detection of Chlamydia trachomatis (CT) and/or Neisseria gonorrhoeae (NG) DNA in male and female urine, clinician-instructed self-collected vaginal swab specimens (collected in a clinical setting), clinician-collected vaginal swab specimens, and endocervical swab specimens, all collected in cobas® PCR Media (Roche Molecular Systems, Inc.), and cervical specimens collected in PreservCyt® solution. This test is intended as an aid in the diagnosis of chlamydial and gonococcal disease in both symptomatic and asymptomatic individuals.

Ancillary Collection Kits 2.1.

The cobas® PCR Media Dual Swab Sample Kit is used to collect and transport endocervical and vaginal swab specimens. The cobas® PCR Media serves as a nucleic acid stabilizing transport and storage medium for gynecological specimens.

Note: This kit has been validated for use with the following tests:

cobas® CT/NG v2.0 Test .
cobas® CT/NG for use on cobas® 6800/8800 Systems .

The cobas® PCR Media Uni Swab Sample Kit is used to collect and transport human specimens. The cobas® PCR Media serves as a nucleic acid stabilizing transport and storage medium for human specimens.

Note: This kit has been validated for use with the following tests:

cobas® CT/NG v2.0 Test .
cobas® CT/NG for use on cobas® 6800/8800 Systems .
cobas® Cdiff Test for use on the cobas® 4800 System .

8

The cobas® PCR Urine Sample Kit is used to collect and transport urine specimens. The cobas® PCR Media serves as a nucleic acid stabilizing transport and storage medium for urine specimens. Use this collection kit only with either cobas® CT/NG on cobas® 6800/8800 Systems or the cobas® CT/NG v2.0 Test.

TECHNOLOGICAL CHARACTERISTICS 3.

The primary technological characteristics and intended use of the RMS cobas® CT/NG for use on the cobas® 6800/8800 systems are substantially equivalent to other legally marketed nucleic acid amplification tests intended for the qualitative detection of Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG).

As indicated in Table 1, the cobas® CT/NG for use on the cobas® 6800/8800 systems is substantially equivalent to significant characteristics of the identified predicate device, the currently cleared cobas® CT/NG v2.0 Test (K163184).

| | Predicate Device:
cobas® CT/NG v2.0 Test
(K163184) | Submitted Device
cobas® CT/NG for use on the
cobas® 6800/8800 systems |
|------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended Use | The cobas® CT/NG v2.0 Test is an
automated, in vitro nucleic acid amplification
test for the qualitative detection of
Chlamydia trachomatis (CT) and/or
Neisseria gonorrhoeae (NG) DNA in
urogenital specimens. The Test utilizes the
Polymerase Chain Reaction (PCR) for the
detection of Chlamydia trachomatis and
Neisseria gonorrhoeae DNA in male and
female urine, self-collected vaginal swab
specimens (collected in a clinical setting),
clinician-collected vaginal swab specimens,
and endocervical swab specimens, all
collected in cobas® PCR Media (Roche
Molecular Systems, Inc.), and cervical
specimens collected in PreservCyt® solution.
This test is intended as an aid in the
diagnosis of chlamydial and gonococcal
disease in both symptomatic and
asymptomatic individuals. | The cobas® CT/NG on the cobas® 6800/8800
system is an automated, qualitative in vitro
nucleic acid diagnostic test, that utilizes real-time
polymerase chain reaction (PCR), for the direct
detection of Chlamydia trachomatis (CT) and/or
Neisseria gonorrhoeae (NG) DNA in male and
female urine, clinician instructed self-collected
vaginal swab specimens (collected in a clinical
setting), clinician-collected vaginal swab
specimens, and endocervical swab specimens,
all collected in cobas® PCR Media (Roche
Molecular Systems, Inc.), and cervical
specimens collected in PreservCyt® solution.
This test is intended as an aid in the diagnosis of
chlamydial and gonococcal disease in both
symptomatic and asymptomatic individuals. |
| | Predicate Device:
cobas® CT/NG v2.0 Test
(K163184) | Submitted Device
cobas® CT/NG for use on the
cobas® 6800/8800 systems |
| | Ancillary Collection Kits: | Ancillary Collection Kits: |
| | The cobas® PCR Media Dual Swab Sample
Kit is used to collect and transport
endocervical and vaginal swab specimens.
The cobas® PCR Media serves as a nucleic
acid stabilizing transport and storage
medium for gynecological specimens. Use
this collection kit only with the cobas®
CT/NG v2.0 Test. | The cobas® PCR Media Dual Swab Sample Kit
is used to collect and transport endocervical and
vaginal swab specimens. The cobas® PCR
Media serves as a nucleic acid stabilizing
transport and storage medium for gynecological
specimens. Note: This kit has been validated for
use with the following tests: cobas® CT/NG v2.0
Test cobas® CT/NG for use on
cobas® 6800/8800 Systems. |
| Intended Use,cont. | The cobas® PCR Media Uni Swab Sample
Kit is used to collect and transport human
specimens. The cobas® PCR Media serves
as a nucleic acid stabilizing transport and
storage medium for human specimens. Use
this collection kit with cobas® CT/NG v2.0
Test and cobas® Cdiff Tests. | The cobas® PCR Media Uni Swab Sample Kit is
used to collect and transport human specimens.
The cobas® PCR Media serves as a nucleic acid
stabilizing transport and storage medium for
human specimens Note: This kit has been
validated for use with the following tests:
cobas® CT/NG v2.0 Test cobas® CT/NG for use
on cobas® 6800/8800 Systems cobas® Cdiff
Test for use on the cobas® 4800 System |
| | The cobas® PCR Urine Sample Kit is used
to collect and transport urine specimens.
The cobas® PCR Media serves as a nucleic
acid stabilizing transport and storage
medium for urine specimens. Use this
collection kit only with the cobas® CT/NG
v2.0 Test. | The cobas® PCR Urine Sample Kit is used to
collect and transport urine specimens. The
cobas® PCR Media serves as a nucleic acid
stabilizing transport and storage medium for
urine specimens Use this collection kit only with
either cobas® CT/NG on cobas® 6800/8800
Systems or the cobas® CT/NG v2.0 Test. |
| Sample Types | • Male and female urine,
• Self-collected/clinician-collected
vaginal swab specimens in
cobas® PCR Media,
• Endocervical swab specimens in
cobas® PCR Media,
• Cervical specimens in PreservCyt®
solution | Same |
| Subject Status | Asymptomatic and symptomatic | Same |
| Sample Collection
Devices | cobas® PCR Media Dual Swab Sample Kit
cobas® PCR Media Uni Swab Sample Kit
cobas® PCR Urine Sample Kit | Same |
| CT Analyte targets | CT cryptic plasmid DNA
CT ompA gene | Same |
| NG Analyte targets | NG genomic DNA | Same |
| | Predicate Device:
cobas® CT/NG v2.0 Test
(K163184) | Submitted Device
cobas® CT/NG for use on the
cobas® 6800/8800 systems |
| Sample
Preparation
Procedure | Semi-automated | Automated |
| Amplification
Technology | Real-time PCR | Same |
| Detection
Chemistry | Paired reporter and quencher fluorescence
labeled probes (TaqMan Technology) using
fluorescence resonance energy transfer
(FRET) | Same |
| Result Analysis | Based on PCR cycle threshold analysis | Same |
| Analyzer | cobas® 4800 System | cobas® 6800/8800 systems |

Comparison of the cobas® CT/NG for use on the cobas® 6800/8800 systems with Table 1: the Predicate Device

9

10

4. NON-CLINICAL PERFORMANCE EVALUATION

4.1. Limit of Detection (LoD)

Analytical sensitivity (Limit of Detection or LoD) was determined by analyzing a dilution series of quantified cultures of Chlamydia trachomatis (serovars D and I) and Neisseria gonorrhoeae isolates 2948 (ATCC 19424) and 891. CT and NG cultures were diluted into a matrix of pooled negative specimens of each sample type and 70-78 replicates were tested for each level in each specimen type. All levels were analyzed across 3 unique lots of reagents. LoD for each specimen type is shown in Table 2 as the target concentration which can be detected in ≥ 95% of the replicates for all lots.

Specimen Types	C. trachomatis		N. gonorrhoeae
	Serovar D		Serovar I		Strain 2948		Strain 891
	LOD
(IFU/mL)	Mean
Ct
Value	LOD
(IFU/mL)	Mean
Ct
Value	LOD
(CFU/mL)	Mean
Ct
Value	LOD
(CFU/mL)	Mean
Ct
Value
Endocervical Swab
in cobas® PCR	0.3	36.6	1.4	37.1	0.4	36.3	0.08	37.5
Vaginal Swab in	0.3	37.3	1.4	37.0	0.4	36.3	0.08	37.0
cobas® PCR	0.2	37.8	1.3	37.1	0.2	36.3	0.04	38.3

Table 2: Analytical sensitivity (Limit of Detection)

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| Cervical Samples
collected into
PreservCyt®

Solution	0.6	37.4	2.9	37.4	0.2	36.7	0.08	37
---------------------------------------------------------------	-----	------	-----	------	-----	------	------	----

IFU = Inclusion Forming Unit; quantification of the same C. trachomatis culture using DFA method equates 1 IFU to 6.6 signal generating units (SGU) for Serovar D, and 13.9 SGU for serovar I, where SGU includes Elementary Bodies as well as Reticulate Bodies of C. trachomatis

CFU = Colony Forming Units

4.2. Inclusivity

Inclusivity testing was performed for 13 additional CT serovars, the Swedish new variant strain (nvCT) and an additional 43 independently isolated strains of NG using one lot of reagents. Testing was performed using CT and NG cultures diluted into pools of negative specimens. Results are shown in Table 3 and Table 4 for CT serovars and NG strains, respectively. Twenty replicates per dilution level were tested for each strain in each specimen type.

	Swab* Specimens		Urine Specimens		PreservCyt Specimens
Serovar Type or Variant	IFU/mL	% Pos	IFU/mL	% Pos	IFU/mL	% Pos
A	1.4	100	0.7	100	1.4	100
B	5.9	100	2.9	100	5.9	100
Ba	18.3	100	9.1	100	18.3	100
C	0.6	100	0.3	100	0.6	100
E	6.4	100	3.2	100	6.4	100
F	3.2	100	1.6	100	3.2	100
G	2.9	100	1.5	100	2.9	100
H	9.7	100	4.8	100	9.7	100
J	1.4	100	0.7	100	1.4	100
K	2.0	100	1.0	100	2.0	100
LGV Type 1	5.9	100	3.0	100	5.9	100
LGV Type 2	12.8	100	6.4	100	12.8	100
LGV Type 3	0.7	100	0.4	100	0.7	100
nyCT	0.7	100	0.3	100	0.7	100

Inclusivity Testing for CT Serovars Table 3:

Vaginal swab samples were used as a representative swab sample type for vaginal and endocervical swab specimens.

12

Numbers of NG Strains	Swab* Specimens
	CFU/mL	% Pos
39	0.4	≥ 95
4	1.0	≥ 95
Total = 43
Numbers of NG Strains	Urine Specimens
	CFU/mL	% Pos
41	0.2	≥ 95
2	0.5	100
Total = 43
Numbers of NG Strains	PreservCyt Specimens
	CFU/mL	% Pos
42	0.4	≥ 95
1	1.0	100
Total = 43

Table 4: Inclusivity Testing for NG strains

Vaginal swab samples were used as a representative swab sample type for vaginal and endocervical swab specimens.

4.3. Precision

In-house precision was examined using a panel composed of CT and NG cultures diluted into a pool of negative endocervical swab specimen matrix collected in cobas® PCR Media, a pool of negative urine matrix plus cobas® PCR Media and a pool of negative cervical specimen matrix collected in PreservCyt® Solution. Endocervical swabs were intended to represent all swab samples collected in cobas® PCR Media (endocervical and vaginal). Four levels were tested using CT serovar D and NG strain 2948 (ATCC 19424) as the target organisms.

The precision panel was designed to include members with high negative, low and moderate concentrations of CT and NG for each panel matrix, corresponding to ~0.3x, ~1x and ~3x LoD. Testing was performed with three lots of cobas® CT/NG reagents and two instruments for a total of 24 runs. A description of the precision panels and the study performance hit rate is shown in Table 5. All negative panel members tested negative throughout the study. Analysis of standard deviation and percent coefficient of variation of the Ct values from valid tests performed on

13

positive panel members (see Table 6 and Table 7) yielded overall CV (%) ranges from 1.62% to 4.05% for CT and from 1.17% to 3.55% for NG. Testing occurred over 12 days, using 2 instruments, with 2 runs per day. Each run consisted of 3 replicates of each sample.

Level	N Tested	N positive CT	N positive NG	Hit Rate		95% CI CT		95% CI NG
				CT	NG	LL	UL	LL	UL
Endocervical Swab in cobas® PCR Media
Negative	72	0	0	0%	0%	0.0	5.0	0.0	5.0
High Negative	72	51	32	71%	44%	59	81	33	57
Low	72	69	68	96%	94%	88	99	86	98
Moderate	72	72	72	100%	100%	95	100	95	100
Cervical samples collected into PreservCyt® Solution
Negative	72	0	0	0%	0%	0.0	5.0	0.0	5.0
High Negative	72	38	47	53%	65%	41	65	53	76
Low	72	72	69	100%	96%	95	100	88	99
Moderate	72	72	72	100%	100%	95	100	95	100
cobas® PCR Media with Urine
Negative	72	0	0	0%	0%	0.0	5.0	0.0	5.0
High Negative	72	56	56	78%	78%	66	87	66	87
Low	72	71	72	99%	100%	92	100	95	100
Moderate	72	72	72	100%	100%	95	100	95	100

Table 5: Summary of within-laboratory precision

14

| Level
(Hit Rate) | Mean Ct | Between
instrument | | Between
lot | | Within run | | Between
run | | Between
day | | Total | |
|------------------------------------------------------|---------|-----------------------|---------|----------------|---------|------------|---------|----------------|---------|----------------|---------|-------|---------|
| | | SD | CV
% | SD | CV
% | SD | CV
% | SD | CV
% | SD | CV
% | SD | CV
% |
| Endocervical Swab in cobas® PCR Media | | | | | | | | | | | | | |
| High
Negative
(71%) | 39.7 | 0.0 | 0.0 | 0.0 | 0.0 | 1.2 | 3.21 | 0.0 | 0.00 | 0.3 | 0.8 | 1.3 | 3.3 |
| Low
(96%) | 38.5 | 0.0 | 0.0 | 0.0 | 0.1 | 1.1 | 2.96 | 0.0 | 0.00 | 0.4 | 1.2 | 1.2 | 3.2 |
| Moderate
(100%) | 36.9 | 0.0 | 0.0 | 0.2 | 0.6 | 0.5 | 1.45 | 0.0 | 0.18 | 0.0 | 0.0 | 0.6 | 1.6 |
| Cervical Samples collected into PreservCyt® Solution | | | | | | | | | | | | | |
| High
Negative
(53%) | 38.3 | 0.6 | 1.5 | 0.5 | 1.3 | 1.1 | 2.92 | 0.0 | 0.00 | 0.0 | 0.0 | 1.3 | 3.5 |
| Low
(100%) | 36.9 | 0.2 | 0.5 | 0.2 | 0.7 | 0.6 | 1.85 | 0.0 | 0.00 | 0.0 | 0.0 | 0.7 | 2.0 |
| Moderate
(100%) | 35.6 | 0.0 | 0.0 | 0.2 | 0.5 | 0.5 | 1.46 | 0.0 | 0.24 | 0.0 | 0.0 | 0.5 | 1.5 |
| cobas® PCR Media with Urine | | | | | | | | | | | | | |
| High
Negative
(78%) | 38.9 | 0.0 | 0.0 | 0.1 | 0.3 | 1.2 | 3.22 | 0.3 | 1.01 | 0.0 | 0.0 | 1.3 | 3.3 |
| Low
(99%) | 38.3 | 0.1 | 0.2 | 0.0 | 0.0 | 1.5 | 3.97 | 0.0 | 0.00 | 0.2 | 0.7 | 1.5 | 4.0 |
| Moderate
(100%) | 37.1 | 0.0 | 0.0 | 0.0 | 0.0 | 1.0 | 2.84 | 0.0 | 0.00 | 0.2 | 0.7 | 1.0 | 2.9 |

Table 6: Overall mean, standard deviations and coefficients of variation (%) for cycle threshold, CT positive panel members

15

| Level
(Hit Rate) | Mea
n Ct | Between
instrumen
t | | Between
lot | | Within run | | Between
run | | Between
day | | Total | |
|------------------------------------------------------|-------------|---------------------------|---------|----------------|---------|------------|---------|----------------|---------|----------------|---------|----------|---------|
| | | SD | CV
% | SD | CV
% | SD | CV
% | SD | CV
% | SD | CV
% | SD | CV
% |
| Endocervical Swab in cobas® PCR Media | | | | | | | | | | | | | |
| High
Negative
(44% ) | 39.1 | 0.0
0 | 0.00 | 0.3
1 | 0.79 | 0.8
4 | 2.14 | 0.7
2 | 1.85 | 0.5
7 | 1.46 | 1.2
8 | 3.28 |
| Low (94%) | 38.1 | 0.0
0 | 0.00 | 0.0
0 | 0.00 | 1.2
7 | 3.34 | 0.0
0 | 0.00 | 0.0
0 | 0.00 | 1.2
7 | 3.34 |
| Moderate
(100%) | 36.5 | 0.0
0 | 0.00 | 0.2
4 | 0.67 | 0.6
9 | 1.89 | 0.0
0 | 0.00 | 0.1
5 | 0.40 | 0.7
4 | 2.04 |
| Cervical Samples collected into PreservCyt® Solution | | | | | | | | | | | | | |
| High
Negative
(65%) | 39.0 | 0.3
4 | 0.87 | 0.0
0 | 0.00 | 1.1
1 | 2.85 | 0.0
8 | 0.20 | 0.4
5 | 1.16 | 1.2
5 | 3.21 |
| Low (96%) | 38.0 | 0.0
0 | 0.00 | 0.0
0 | 0.00 | 1.2
5 | 3.28 | 0.0
0 | 0.00 | 0.0
0 | 0.00 | 1.2
5 | 3.28 |
| Moderate
(100%) | 35.8 | 0.0
0 | 0.00 | 0.2
8 | 0.78 | 0.7
6 | 2.13 | 0.0
0 | 0.00 | 0.0
0 | 0.00 | 0.8
1 | 2.27 |
| cobas® PCR Media with Urine | | | | | | | | | | | | | |
| High
Negative
(78%) | 39.1 | 0.0
0 | 0.00 | 0.2
6 | 0.66 | 1.3
5 | 3.46 | 0.0
0 | 0.00 | 0.1
8 | 0.45 | 1.3
9 | 3.55 |
| Low
(100%) | 36.7 | 0.1
4 | 0.38 | 0.1
6 | 0.42 | 0.7
1 | 1.92 | 0.0
0 | 0.00 | 0.0
0 | 0.00 | 0.7
4 | 2.00 |
| Moderate
(100%) | 34.9 | 0.0
0 | 0.00 | 0.1
6 | 0.47 | 0.3
7 | 1.06 | 0.0
6 | 0.18 | 0.0
0 | 0.00 | 0.4
1 | 1.17 |

Table 7: Overall mean, standard deviations and coefficients of variation (%) for cycle threshold, NG positive panel members

Analytical specificity/Cross-reactivity 4.4.

A panel of 149 bacteria, fungi and viruses, including those commonly found in the male and female urogenital tract, 20 representatives of non-gonorrhoeae Neisseria strains and other phylogenetically unrelated organisms, were tested with cobas®CT/NG to assess analytical specificity. The organisms listed in Table 8 were spiked at concentrations of approximately 1 x 106 units*/mL for bacteria and approximately 1 x 105 units*/mL for viruses into pools of negative vaginal swab specimens in cobas® PCR Media, urine stabilized in cobas® PCR Media

16

and cervical specimens in PreservCyt® Solution. Testing was performed with each potential interfering organism alone as well as with each organism mixed with CT and NG cultures at ~3x LoD. Results indicated that none of these organisms interfered with the detection of CT and NG or produced false positive results in the CT/NG negative matrices. (N=3 across the tested specimen types).

*All bacteria were quantified as Colony Forming Units (CFU) except Chlamydophila pneumonia and Chlamydophila psittaci which were quantified as Elementary Bodies (EB). All viruses were quantified as units/mL as determined by TCID50 Endpoint Dilution Assay. Trichomonas vaginalis and HPV16 were quantified as cells/mL.

17

Achromobacter xerosis	Gemella haemolysans	Neisseria subflava
Acinetobacter calcoaceticus	Haemophilus ducreyi	Neisseria weaverii
Acinetobacter Iwoffi	Haemophilus influenzae	Pantoea agglomerans
Actinomyces israelii	Helicobacter pylori	Paracoccus denitrificans
Aerococcus viridans	Herpes simplex virus I	Peptostreptococcus anaerobius
Aeromonas hydrophila	Herpes simplex virus II**	Peptostreptococcus
asaccharolyticus
Alcaligenes faecalis	HPV16*	Peptostreptococcus magnus
Atopobium vaginae	Kingella dentrificans	Plesiomonas shigelloides
Bacillus subtilis	Kingella kingae	Propionibacterium acnes
Bacteriodes fragilis	Klebsiella oxytoca	Proteus mirabilis
Bacteroides caccae	Klebsiella pneumoniae	Proteus penneri
Bacteroides ureolyticus	Lactobacillus acidophillus	Proteus vulgaris
Bergeriella denitrificans	Lactobacillus brevis	Providencia rettgeri
Bifidobacterium adolescentis	Lactobacillus crispatus	Providencia stuartii
Bifidobacterium breve	Lactobacillus jensenii	Pseudomonas aeruginosa
Bifidobacterium longum	Lactobacillus lactis	Pseudomonas fluorescens
Blautia product	Lactobacillus leichmannii	Pseudomonas putida
Branhamella catarrhalis	Lactobacillus oris	Rahnella aquatilis
Brevibacterium linens	Lactobacillus parabuchnerri	Rhizobium radiobacter
Campylobacter coli	Lactobacillus reuteri	Rhodospirillum rubrum
Campylobacter jejuni	Lactobacillus vaginalis	Saccharomyces cerevisiae
Candida albicans	Lactococcus lactis cremoris	Salmonella choleraesuis
Candida glabrata	Legionella pneumophila	Salmonella minnesota
Candida parapsilosis	Leuconostoc paramesenteroides	Salmonella typhimurium
Candida tropicalis	Listeria monocytogenes	Serratia denitrificans
Chlamydophila pneumoniae	Micrococcus luteus	Serratia marcescens
Chlamydophila psittaci	Moraxella lacunata	Shigella dysenteriae
Chromobacter violaceum	Moraxella osloensis	Staphylococcus aureus
Citrobacter freundii	Morganella morganii	Staphylococcus epidermidis
Clostridium difficile	Mycobacterium smegmatis	Staphylococcus saprophyticus
Clostridium perfringens	Mycoplasma genitalium***	Streptococcus agalactiae
Corynebacterium genitalium	Mycoplasma hominis	Streptococcus anginosus
Corynebacterium xerosis	Neisseria cinerea	Streptococcus bovis
Cryptococcus neoformans	Neisseria elongata subsp. elongata	Streptococcus dysgalactiae
Cytomegalovirus**	Neisseria elongata subsp. nitroreducens	Streptococcus equinis
Deinococcus radiodurans	Neisseria flava	Streptococcus mitis

Derxia gummosa	Neisseria flavescens	Streptococcus mutans
Eikenella corrodens	Neisseria kochi	Streptococcus pneumoniae
Enterobacter aerogenes	Neisseria lactamica	Streptococcus pyogenes
Enterobacter cloacae	Neisseria macacae	Streptococcus salivarius
Enterococcus avium	Neisseria meningitidis Serogroup A	Streptococcus sanguis
Enterococcus casseliflavus	Neisseria meningitidis Serogroup B	Streptomyces griseinus
Enterococcus faecalis	Neisseria meningitidis Serogroup C	Trichomonas vaginalis
Enterococcus faecium	Neisseria meningitidis Serogroup D	Trueperella pyogenes
Erysipelothrix rhusiopathiae	Neisseria meningitidis Serogroup W135	Ureaplasma urealyticum
Escherichia coli	Neisseria meningitidis Serogroup Y	Veillonela parvula
Escherichia fergusonii	Neisseria mucosa	Vibrio cholerae
Flavobacterium meningosepticum	Neisseria perflava	Vibrio parahaemolyticus
Fusobacterium nucleatum	Neisseria polysaccharea	Yersinia enterocolitica
Gardnerella vaginalis	Neisseria sicca	-

Table 8: Microorganisms tested for analytical specificity/cross reactivity

18

HPV16 was tested as CaSki cells.

** Organism was tested at a concentration of 1 x 104 Units/mL.

***Organism was tested at a concentration of 1 x 105 CFU/mL.

4.5. Interference

The effects of over-the-counter or prescription products that may be present in urogenital specimens (Table 9), were evaluated. Testing was done using pooled clinical specimens (vaginal swab, urine and PreservCyt® specimens) with spiking of potential interferents at levels expected from normal patient usage. Interferents were tested in CT/NG negative specimen pools as well as in specimen pools with CT/NG at ~3x LoD. CT serovars D and I and NG strains 2948 (ATCC 19424) and 891 were used in this study. Five replicates each of CT/NG negative and CT/NG positive samples were tested with each product in each specimen type, except for RepHresh™ Odor Eliminating Vaginal Gel and RepHresh™ Clean Balance Gel, which were tested with 2 replicates each to verify interference that had been observed with Replens™ Long-Lasting Vaginal Moisturizer, a product with a similar formulation.

Of the over-the-counter (OTC) and prescription products tested, Metronidazole Vaginal Gel, Replens™ Long-Lasting Vaginal Moisturizer, RepHresh™ Odor Eliminating Vaginal Gel and RepHresh™ Clean Balance produced false negative or invalid results in at least one replicate of the samples tested.

19

| Product Name | Vaginal
Swabs | Urine | PreservCyt
Solution |
|------------------------------------------------------------------------------|------------------|-------|------------------------|
| | mg/mL | mg/mL | mg/mL |
| Clindamycin Phosphate Vaginal Cream | 7.1 | 3.4 | 1.6 |
| Equate tioconazole 1 | 3.7 | 1.7 | 0.8 |
| Equate Vagicaine Anti-Itch Cream | 4.1 | 2.0 | 0.9 |
| Estrace | 3.8 | 2.0 | 1.0 |
| K-YTM Ultra Gel | 5.7 | 2.7 | 1.2 |
| Metronidazole Vaginal Gel | 0.1* | 0.1* | 0.2* |
| Monistat 3 Vaginal Antifungal Combination Pack | 3.7 | 1.7 | 0.7 |
| Monistat® Complete Care Itch Relief Cream | 3.7 | 1.8 | 0.9 |
| 7 Day Vaginal Cream | 3.9 | 1.8 | 0.8 |
| Norforms Suppositories | 3.4 | 1.7 | 0.7 |
| Premarin | 6.1 | 3.1 | 1.4 |
| ReplensTM Long-Lasting Vaginal Moisturizer | 0.05* | 0.05* | 0.2* |
| Summer's Eve Feminine Deodorant Spray | 6.4 | 3.1 | 2.0 |
| VCF - Vaginal Contraceptive Foam | 2.1 | 1.0 | 0.4 |
| Yeast Gard Advanced | 3.7 | 1.7 | 1.0 |
| Azo Standard (urine only) | N/A | 0.1 | N/A |
| RepHreshTM Odor Eliminating Vaginal Gel | ‡ | ‡ | ‡ |
| RepHreshTM Clean Balance Gel | ‡ | ‡ | ‡ |
| * Concentrations above this level may cause interference in clinical samples | | | |

Table 9: List of substances with concentrations tested that do not interference with test performance in urogenital specimens

RepHresh™ products were tested using simulated swab specimen. Concentrations of product that did not interfere with test performance were not determined

Endogenous substances that may be present in urogenital specimens were tested for interference. Testing was done using pooled clinical specimens (endocervical swab, urine and PreservCyt® specimens) with spiking of potential endogenous interferents were tested in CT/NG negative specimen pools as well as in the presence of CT/NG at ~3x LoD. CT serovars D and I and NG strains 2948 (ATCC 19424) and 891 were used in this study. Five replicates each of CT/NG negative and CT/NG positive samples were tested with each substance in each specimen type.

20

Interference was noted with whole blood at 10% for urine and PreservCyt® specimens and with cervical mucus at 1% in endocervical specimens when at least one replicate of the samples tested produced false negative or invalid results . Levels of endogenous substances tolerated by the assay for all specimen types are shown in Table 10.

Interferent	Endocervical Swab	PreservCyt®	Urine
Albumin (% w/v)	N/A	N/A	5%
Bilirubin (% w/v)	N/A	N/A	0.5%
Mucus (% w/v)	0.5%	1.0%	0.5%
Glucose (% w/v)	N/A	N/A	1.0%
Peripheral Blood Mononuclear Cells
(PBMCs as cells/mL)	1.0E+06	1.0E+06	1.0E+06
pH (acidic and alkaline)	N/A	N/A	pH 4 and pH 9
Semen (% w/v)	1.5%	1.5%	N/A
Whole Blood (% v/v)	10%	5%	5%

Table 10: Summary of endogenous substance concentrations that do not show interference

4.6. Competitive Inhibition

To assess competitive inhibition between CT and NG, samples of vaginal swab, urine and PreservCyt® specimens were tested with low and moderate concentrations of one target mixed with very high concentrations of the opposite target. Low and moderate concentrations were defined as ~1x LoD and ~3x LoD, respectively, and high concentrations (≥103 IFU/mL for CT and ≥104 CFU/mL for NG) were defined as generating a signal greater than observed in 95% of target positive clinical specimens. Testing results indicated that when NG was present at a high concentration, CT was detected in all specimen types, at both low (~1x LoD) and moderate (~3x LoD) levels. Results also indicated that when CT was present at a high concentration, NG was detected in all specimen types at moderate (~3x LoD) levels, however, NG was not consistently detected at low levels (Expanded testing indicated detection in 35% (7/20) of the samples at 0.4 CFU/ml and 60% (12/20) of the samples at 0.65 CFU/ml).

21

Cross contamination/Carryover 4.7.

Studies were performed to evaluate potential cross-contamination on the cobas® 6800/8800 Systems using cobas® CT/NG. Cross-contamination can cause false positive results. In this performance study the sample-to-sample cross-contamination rate of cobas® CT/NG has been determined to be 0.5% (2/432), (95% CI: 0.1%-1.7%) when alternating very high positive and negative samples were tested over nine runs. Run-to-run cross-contamination has not been observed (0/282). Testing was done using samples prepared with cobas® PCR Media and with PreservCyt® Solution and with urine stabilized in cobas® PCR Media. High positive samples (≥103 IFU/mL for CT and ≥104 CFU/mL for NG) in the study were prepared to generate a Ct value that was lower than that obtained with 95% or more of the specimens of infected patients in the intended use population. Cross contamination rates in clinical settings depend on the proportion of high positive samples and prevalence of the disease. Routine clinical crosscontamination rates are expected to be lower than what was observed in this study and need to be assessed in user's settings.

CLINICAL PERFORMANCE EVALUATION 5.

Clinical Performance 5.1.

5.1.1. Study Design

The clinical utility and performance of cobas® CT/NG was established in a multi-site, prospective collection study by comparing the results to a Patient Infected Status (PIS) that used a combination of FDA-cleared NAATs for urogenital specimens. Female and male urogenital specimens were collected at 9 geographically diverse sites in the US with testing performed at 4 laboratory testing sites (3 external and 1 internal).

Prospectively enrolled female subjects provided the following urogenital specimens: first-void urine, 3 vaginal swabs, 1 endocervical swab in cobas® PCR Media, and 1 cervical sample in PreservCyt® Solution. If the female was in the clinician-collected vaginal swab arm of the study, 2 of the vaginal swabs were placed in the respective manufacturers' collection device and 1 in cobas® PCR Media. If the female subject was in the self-collected vaginal swab self-collection arm of the study, then 1 vaginal swab was self-collected first and placed into cobas® PCR Media

22

and then followed by the 2 clinician-collected vaginal swabs and placed in the 2 respective manufacturers' collection devices.

Prospectively enrolled male subjects provided a urine specimen that was aliquoted into the respective manufacturers' collection device and cobas® PCR Media.

Subjects were classified as symptomatic if they self-reported symptoms indicative of a CT or NG infection as listed below:

Dysuria (pain during urination) .
Coital pain, difficulty or bleeding .
Pelvic pain .
Abnormal vaginal discharge .
Pelvic, uterine or ovarian pain .
Urethral discharge .
Testicular pain .
Scrotal pain or swelling .

Prospectively enrolled subjects were classified as asymptomatic if they did not report any of the above symptoms.

Specimens were tested for CT and NG using cobas® CT/NG and commercially available FDAcleared NAATs. All tests were run according to the respective manufacturers' Instructions For Use.

The clinical performance of cobas® CT/NG was evaluated by comparing the results from collected specimen types to a pre-specified PIS (Patient Infected Status) algorithm as determined by the combined results from 2 commercially available NAATs for females and 3 commercially available NAATs for males. The PIS algorithms for Female and Male subjects are shown in Table 11 and Table 12, respectively.

23

For NG, archived prospectively collected female urine, cervical specimens in PreservCyt®, and endocervical swabs were obtained from the clinical study for cobas® CT/NG v2 test on the cobas® 4800 System. The PIS of these specimens were already determined from the clinical study for cobas® CT/NG v2 test on the cobas® 4800 System..

| NAAT1
Urine/Vaginal | NAAT2
Urine/Vaginal | Patient Infected Status (PIS)b |
|------------------------|------------------------|--------------------------------------------|
| +/+ | +/+ | Infected |
| +/+ | +/- or -/+ | Infected |
| +/- or -/+ | +/+ | Infected |
| +/- | -/+ | Infected |
| -/+ | +/- or -/+ | Infected |
| +/- | +/- | Infected (Urine)
Non-Infected (Vaginal) |
| +/- or -/+ | -/- | Not Infected |
| +/+ | -/- | Not Infected |
| -/- | +/+ | Not Infected |
| -/- | +/- or -/+ | Not Infected |
| -/- | -/- | Not Infected |

Table 11: Determination of female Patient Infected Status (PIS) for urogenital specimensª

ª One or more positives in each NAAT (NAAT1 and NAAT2) designates the PIS as Infected. Any other combination of results defines the PIS as Not Infected.

b In the scenario where one or more of the sample types are invalid, the remaining sample types with valid results from NAAT1 and NAAT2 must have concordant positive or concordant negative results to determine the PIS as Infected or Not Infected, respectively. For all other cases where one or more of the sample types are invalid, the PIS is indeterminate.

24

| NAAT1
Urine | NAAT2
Urine | NAAT3
Urine | Patient Infected Status
(PIS)a |
|----------------|----------------|----------------|-----------------------------------|
| + | + | + | Infected |
| + | + | - | Infected |
| + | - | + | Infected |
| - | + | + | Infected |
| - | - | + | Not Infected |
| - | + | - | Not Infected |
| + | - | - | Not Infected |
| - | - | - | Not Infected |

Table 12: Determination of male Patient Infected Status (PIS) for urine specimens

ª If at least 2 out of the 3 test results are concordant positive then the PIS can be considered as Infected or Not Infected, respectively. If one test result is invalid/missing and the other two test results are discordant then the PIS is indeterminate. If 2 or 3 test results are invalid/missing, then the PIS is indeterminate.

Sensitivity (SENS), specificity (SPEC), positive predictive value (PPV), and negative predictive value (NPV) of cobas® CT/NG were calculated separately for the detection of CT or NG using the PIS as the composite reference standard and evaluated by gender, sample type, and symptom status. In addition, the predictive values were calculated based on overall sensitivity and specificity (with all data combined for males and females) for a range of hypothetical prevalence values.

5.1.2. Results

A total of 5.197 subjects were prospectively enrolled, of which 5.105 were eligible for inclusion. Of the 5,105 eligible subjects contributing prospective specimens, 5,053 (99.0%) (3,860 females and 1,193 males) were evaluable and were included in the data analyses. A total of 52 subjects (1.0%) were classified as non-evaluable and excluded from all statistical analyses. There were a total of 371 archived prospectively collected female urogenital samples (urine, cervical specimens in PreservCyt, and endocervical swabs) tested in this clinical study from 295 female subjects. Among the 17.169 samples tested in this study, 19 samples exhibited invalid results on the first run (invalid rate of 0.11% (95%CI: 0.07% ; 0.17%)). Upon repeat testing, 3 samples exhibited valid results.

25

Table 13 and Table 14 summarize the results from symptomatic and asymptomatic, prospectively enrolled subjects designated as infected or non-infected with CT (females and males, respectively) according to the PIS algorithm. A total of 271 females and 118 males were infected with CT. Symptoms were reported in 45.8% (124/271) of infected and 36.7% (1318/3589) of non-infected females. Symptoms were reported in 53.4% (63/118) of infected and 22.5% (242/1074) of non-infected males.

26

| | Statusa | | |
|-------------------------------|-- | Patient
Infected
Status | | UR | VS | Infected | | Infected | | Infected | | Infected | | Infected | | Infected | | Infected | | Infected | | Infected | | Infected | | Infected | | Infected | | Infected | | Infected | | Infected | | Infected | | Infected | | Infected | | Infected | | Infected | | Infected | | Infected | | Infected | | Infectedb | | Infectedb | | Infectedb | | Total | | Non-Infected | | Non-Infected | | Non-Infected | | Non-Infected | | Non-Infected | | Statusª | | | Patient
Infected
Status | UR | VS | UR | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Statusa | | | Patient
Infected
Status | UR | VS | UR | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Non-Infected | Total | | NAAT1 | | NAAT2 | | cobas® CT/NG | | | | Symptom
-------|---------|---------|---------|--------------|--------------|--------------------|---------|--------------------|--------------------|-------|-------|
| UR | VS | UR | VS | PC | ES | Symp | Asymp | Total |
| + | + | + | + | + | + | + | + | 104 | 108 | 212 |
| - | + | + | + | + | + | + | + | 2 | 7 | 9 |
| + | + | + | + | + | + | - | + | 1 | 5 | 6 |
| + | + | + | + | + | + | + | + | 2 | 4 | 6 |
| + | + | + | + | + | + | - | - | 1 | 4 | 5 |
| + | + | + | + | + | + | + | - | 1 | 3 | 4 |
| - | + | + | + | - | + | + | + | 1 | 3 | 4 |
| - | + | - | + | - | + | + | + | 2 | 2 | 4 |
| - | + | - | + | + | + | + | + | 2 | 1 | 3 |
| + | - | + | + | + | + | - | - | 1 | 1 | 2 |
| + | + | + | + | + | + | Failed | + | 0 | 1 | 1 |
| + | + | + | + | + | + | + | Failed | 1 | 0 | 1 |
| - | + | + | + | + | + | + | + | 0 | 1 | 1 |
| - | + | + | + | + | + | + | - | 0 | 1 | 1 |
| - | + | + | + | + | + | - | + | 1 | 0 | 1 |
| - | + | + | + | + | + | - | - | 0 | 1 | 1 |
| - | + | + | + | - | + | - | - | 1 | 0 | 1 |
| - | + | - | + | + | + | + | - | 0 | 1 | 1 |
| - | + | - | + | - | + | - | + | 0 | 1 | 1 |
| + | - | + | + | + | + | + | + | 1 | 0 | 1 |
| + | - | + | + | + | - | - | - | 0 | 1 | 1 |
| + | - | - | + | - | + | - | - | 1 | 0 | 1 |
| + | - | - | + | - | + | | | 0 | 1 | 1 |
| + | - | + | - | + | + | - | + | 1 | 0 | 1 |
| + | - | + | - | + | + | - | - | 1 | 0 | 1 |
| + | - | + | - | + | - | - | - | 0 | 1 | 1 |
| | | | | | | | | 124 | 147 | 271 |
| - | - | - | - | - | - | - | - | 1252 | 2165 | 3417 |
| - | - | - | + | - | - | - | - | 6 | 12 | 18 |
| - | Invalid | - | - | - | - | - | - | 7 | 5 | 12 |
| - | - | - | - | - | Invalid | | - | 6 | 4 | 10 |
| - | - | - | - | - | + | - | - | 1 | 9 | 10 |
| NAAT1 | | | NAAT2 | cobas® CT/NG | | Symptom
| | | |
| VS | UR | VS | PC | ES | Symp | Asymp | Total | |
| - | - | + | - | - | - | - | - | 2 | 7 | 9 | |
| - | - | - | - | - | - | - | + | 5 | 4 | 9 | |
| - | - | NA | - | - | - | - | - | 2 | 7 | 9 | |
| - | - | Invalid | - | - | - | - | - | 0 | 9 | 9 | |
| - | - | - | - | - | - | - | NA | 3 | 5 | 8 | |
| + | - | - | - | - | - | - | - | 3 | 3 | 6 | |
| - | - | - | - | + | - | - | - | 1 | 5 | 6 | |
| - | + | - | - | - | - | - | - | 2 | 2 | 4 | |
| - | - | - | - | - | - | + | - | 1 | 3 | 4 | |
| - | - | - | - | - | NA | - | - | 1 | 3 | 4 | |
| - | NA | - | - | - | - | - | - | 0 | 4 | 4 | |
| - | - | - | - | - | - | NA | NA | 0 | 3 | 3 | |
| - | - | - | NA | - | - | - | - | 1 | 2 | 3 | |
| NA | - | - | - | - | - | - | - | 0 | 3 | 3 | |
| Invalid | - | - | - | - | - | - | - | 2 | 1 | 3 | |
| - | + | - | - | - | + | - | - | 2 | 0 | 2 | |
| - | - | - | + | - | + | + | + | 0 | 2 | 2 | |
| - | - | - | + | - | + | + | - | 2 | 0 | 2 | |
| - | - | - | + | - | + | - | + | 1 | 1 | 2 | |
| - | - | - | + | - | + | - | - | 0 | 2 | 2 | |
| - | - | - | - | - | + | + | + | 2 | 0 | 2 | |
| - | - | - | - | - | - | - | Invalid | 2 | 0 | 2 | |
| - | - | - | Invalid | - | - | - | - | 1 | 1 | 2 | |
| - | - | + | + | - | + | - | - | 0 | 1 | 1 | |
| - | - | + | - | + | - | - | - | 0 | 1 | 1 | |
| - | - | + | - | - | + | - | - | 1 | 0 | 1 | |
| - | - | - | + | + | + | + | + | 0 | 1 | 1 | |
| - | - | - | + | - | - | + | + | 1 | 0 | 1 | |
| - | - | - | + | + | + | - | + | 0 | 1 | 1 | |
| - | - | - | + | - | - | - | + | 0 | 1 | 1 | |
| - | - | - | + | + | + | - | - | 0 | 1 | 1 | |
| - | - | - | - | - | - | NA | - | 0 | 1 | 1 | |
| - | - | - | - | - | Invalid | Invalid | Invalid | 1 | 0 | 1 | |
| - | - | - | - | - | - | Invalid | Invalid | 1 | 0 | 1 | |
| NAAT1 | | NAAT2 | | cobas® CT/NG | | | | Symptom
| |
| VS | UR | VS | PC | ES | Symp | Asymp | Total | |
| - | - | - | - | - | NA | Invalid | - | 1 | 0 | 1 | |
| - | - | - | - | - | - | Invalid | - | 1 | 0 | 1 | |
| - | - | - | - | - | + | + | - | 1 | 0 | 1 | |
| - | - | - | - | - | - | - | Failed | 1 | 0 | 1 | |
| - | - | - | - | + | - | - | + | 1 | 0 | 1 | |
| - | - | - | - | - | + | - | + | 1 | 0 | 1 | |
| - | - | - | - | Failed | - | - | - | 1 | 0 | 1 | |
| - | - | - | - | - | Failed | - | - | 1 | 0 | 1 | |
| - | - | - | Invalid | - | + | + | + | 0 | 1 | 1 | |
| - | - | NA | + | - | + | - | - | 0 | 1 | 1 | |
| - | Invalid | - | - | - | Invalid | - | - | 1 | 0 | 1 | |
| | | | | | | | 1318 | 2271 | 3589 | |

Table 13: CT positive/negative analyses for female Patient Infected Status

27

28

ª Symp = symptomatic, Asymp = asymptomatic.

b Infected (Urine), Non-Infected (Swabs).

Note: In the scenario where one or more of the sample types are invalidinot available (NA), for female subjects, the remaining sample types with valid results from NAAT2 must have concordant positive or concordant negative results to determine the PIS as Infected or Not Infected, respectively. For all other cases where one or more of the sample types are invalid/not available (NA), the PIS is indeterminate.

Note: Female subjects with designated infection status (Infected) and final valid cobas® CT/NG test results are considered evaluable and included in this summary table.

Note: + denotes Positive, - denotes Negative, NA denotes Not Available.

Note: UR = urine, VS = vaginal swab, PC = PreservCyt®, ES = endocervical swab.

Note: cobas Invalid are the sum of instrument amplification errors and samples excluded due to protocol deviations

Note: cobas Failed are hardware, software or operator errors causing no result reported

29

| | NAAT1 | NAAT2 | NAAT3 | cobas®
CT/NG | Symptom Statusa | | |
|-------------------------------|---------|---------|---------|-----------------|-----------------|-------|-------|
| Patient
Infected
Status | UR | UR | UR | UR | Symp | Asymp | Total |
| Infected | + | + | + | + | 60 | 55 | 115 |
| Infected | - | + | + | + | 1 | 0 | 1 |
| Infected | + | Invalid | + | + | 1 | 0 | 1 |
| Infected | + | - | + | + | 1 | 0 | 1 |
| Total Infected | | | | | 63 | 55 | 118 |
| Non-Infected | - | - | - | - | 238 | 819 | 1057 |
| Non-Infected | - | Invalid | - | - | 2 | 2 | 4 |
| Non-Infected | Invalid | - | - | - | 0 | 3 | 3 |
| Non-Infected | - | - | Invalid | - | 0 | 3 | 3 |
| Non-Infected | NA | - | - | - | 1 | 1 | 2 |
| Non-Infected | - | - | - | + | 0 | 2 | 2 |
| Non-Infected | - | - | + | - | 0 | 1 | 1 |
| Non-Infected | - | + | - | + | 1 | 0 | 1 |
| Non-Infected | + | - | - | - | 0 | 1 | 1 |
| Total Non-
Infected | | | | | 242 | 832 | 1074* |

Table 14: CT positive/negative analysis for male Patient Infected Status

a Symp = symptomatic, Asymp = asymptomatic.

*One subject was missing symptom status and is not presented in this table.

Note: If at least 2 out of the 3 test results, for male subjects, are concordant positive or negative then the PIS can be considered as infected or non-infected, respectively. If one test result is invalid/not available (NA) and the other two test results are discordant then the PIS is indeterminate. If 2 or 3 test results are invalid/not available, then the PIS is indeterminate.

Note: Male subjects with designated patient infection status (Infected or Non-Infected) and final valid cobas® CT/NG test results are considered evaluable and included in this summary table.

Note: cobas Invalid are the sum of instrument amplification errors and samples excluded due to protocol deviationsNote: + denotes Positive, - denotes Negative, NA denotes Not Available.

Note: UR = urine.

Sensitivity, specificity, and predictive values of cobas® CT/NG for CT as defined by PIS are presented by gender, sample type, and symptom status in Table 15.

30

| Sample
Typeª | Symptom
Statusb | Total
(n) | SENS | 95% Score
CI | SPEC | 95% Score
CI | PREV
(%) | PPV
(%) | NPV
(%) |
|-----------------|--------------------|--------------|---------------------|--------------------|----------------------|--------------------|-------------|------------|------------|
| | Female | | | | | | | | |
| UR | Symp | 1441 | 96.0%
(119/124) | (90.9%,
98.3%) | 99.8%
(1315/1317) | (99.4%,
100.0%) | 8.6 | 98.3 | 99.6 |
| | Asymp | 2418 | 95.2%
(140/147) | (90.5%,
97.7%) | 99.6%
(2262/2271) | (99.2%,
99.8%) | 6.1 | 94.0 | 99.7 |
| | Overall | 3859 | 95.6%
(259/271)c | (92.4%,
97.4%) | 99.7%
(3577/3588) | (99.5%,
99.8%) | 7.0 | 95.9 | 99.7 |
| VS-C | Symp | 711 | 100.0%
(63/63) | (94.3%,
100.0%) | 99.2%
(643/648) | (98.2%,
99.7%) | 8.9 | 92.6 | 100.0 |
| | Asymp | 1225 | 97.6%
(83/85) | (91.8%,
99.4%) | 99.0%
(1129/1140) | (98.3%,
99.5%) | 6.9 | 88.3 | 99.8 |
| | Overall | 1936 | 98.6%
(146/148) | (95.2%,
99.6%) | 99.1%
(1772/1788) | (98.6%,
99.4%) | 7.6 | 90.1 | 99.9 |
| VS-S | Symp | 720 | 100.0%
(59/59) | (93.9%,
100.0%) | 98.8%
(653/661) | (97.6%,
99.4%) | 8.2 | 88.1 | 100.0 |
| | Asymp | 1186 | 98.4%
(60/61) | (91.3%,
99.7%) | 99.2%
(1116/1125) | (98.5%,
99.6%) | 5.1 | 87.0 | 99.9 |
| | Overall | 1906 | 99.2%
(119/120) | (95.4%,
99.9%) | 99.0%
(1769/1786) | (98.5%,
99.4%) | 6.3 | 87.5 | 99.9 |
| РС | Symp | 1438 | 95.1%
(116/122) | (89.7%,
97.7%) | 99.5%
(1309/1316) | (98.9%,
99.7%) | 8.5 | 94.3 | 99.5 |
| | Asymp | 2413 | 90.3%
(131/145) | (84.4%,
94.2%) | 99.7%
(2261/2268) | (99.4%,
99.9%) | 6.0 | 94.9 | 99.4 |
| | Overall | 3851 | 92.5%
(247/267) | (88.7%,
95.1%) | 99.6%
(3570/3584) | (99.3%,
99.8%) | 6.9 | 94.6 | 99.4 |
| ES | Symp | 1433 | 95.9%
(116/121) | (90.7%,
98.2%) | 99.1%
(1300/1312) | (98.4%,
99.5%) | 8.4 | 90.6 | 99.6 |
| | Asymp | 2410 | 91.1%
(133/146) | (85.4%,
94.7%) | 99.5%
(2253/2264) | (99.1%,
99.7%) | 6.1 | 92.4 | 99.4 |
| | Overall | 3843 | 93.3%
(249/267) | (89.6%,
95.7%) | 99.4%
(3553/3576) | (99.0%,
99.6%) | 6.9 | 91.5 | 99.5 |
| | Male | | | | | | | | |
| UR | Symp | 305 | 100.0%
(63/63) | (94.3%,
100.0%) | 99.6%
(241/242) | (97.7%,
99.9%) | 20.7 | 98.4 | 100.0 |
| | Asymp | 887 | 100.0%
(55/55) | (93.5%,
100.0%) | 99.8%
(830/832) | (99.1%,
99.9%) | 6.2 | 96.5 | 100.0 |
| | Overall | 1192* | 100.0%
(118/118) | (96.8%,
100.0%) | 99.7%
(1071/1074) | (99.2%,
99.9%) | 9.9 | 97.5 | 100.0 |

Table 15: CT clinical performance compared with Patient Infected Status by gender, sample type, and symptom status

31

-----------------	--------------------	--------------	------	-----------------	------	-----------------	-------------	------------	------------

a UR = urine, VS-C = clinician-collected vaginal swab, VS-S = self-collected vaginal swab, PC = PreservCyt®, ES = endocervical swab.

b Symp = symptomatic, Asymp = asymptomatic.

^ Five CT PIS infected females had a CT negative urine specimen with NAAT1 while they had a CT positive vaginal swab with NAAT1 and NAAT2.

One subject was missing symptom status and is not presented in this table.

Note: In the scenario where one or more of the sample types are invalid/not available, for female subjects, the remaining sample types with valid results from NAAT2 must have concordant positive or concordant negative results to determine the PIS as Infected, respectively. For all other cases where one or more of the sample types are invalid/not available, the PIS is indeterminate.

Note: If at least 2 out of the 3 test results, for male subjects, are concordant positive or negative then the PIS can be considered as infected or non-infected, respectively. If one test result is invalid/not available and the other two test results are discordant then the PIS is indeterminate. If 2 or 3 test results are invalid/not available, then the PIS is indeterminate.

Note: Subjects with designated patient infection status (Infected) and final valid cobas® CT/NG test results are considered evaluable and included in this summary table. An evaluable subject may not have all available sample types or valid test results.

Note: Cl = confidence interval, PREV = prevalence, SENS = sensitivity, SPEC = specificity, PPV = positive predictive value, NPV = negative predictive value.

Table 16 and Table 17 summarize the results from symptomatic and asymptomatic subjects

designated as infected or non-infected with NG (females and males, respectively) according to

the PIS algorithm. A total of 57 females and 87 males were infected with NG. Symptoms were

reported in 45.6% (26/57) of infected and 37.2% (1416/3803) of non-infected females.

Symptoms were reported in 94.3% (82/87) of infected and 20.2% (223/1105) of non-infected males.

32

	NAAT1		NAAT2		cobas® CT/NG			Symptom Statusa
Patient
Infected
Status	UR	VS	UR	VS	UR	VS	PC	ES	Symp	Asymp	Total
Infected	+	+	+	+	+	+	+	+	20	23	43
Infected	-	+	-	+	-	+	+	+	2	3	5
Infected	+	+	-	+	+	+	+	+	0	2	2
Infected	-	+	-	+	+	+	+	+	2	0	2
Infected	+	+	+	+	+	+	+	Failed	1	0	1
Infected	+	+	+	+	+	+	-	-	0	1	1
Infected	+	+	-	+	-	+	-	-	0	1	1
Infected	-	+	NA	+	+	+	+	-	0	1	1
Infectedb	+	-	+	-	+	-	-	-	1	0	1
Total									26	31	57

Table 16: NG positive/negative analysis for female Patient Infected Status (prospective specimens)

		NAAT1		NAAT2		cobas® CT/NG				Symptom Statusa
Patient
Infected
Status	UR	VS	UR	VS	UR	VS	PC	ES	Symp	Asymp	Total
Non-Infected	-	-	-	-	-	-	-	-	1368	2315	3683
Non-Infected	-	+	-	-	-	-	-	-	4	11	15
Non-Infected	+	-	-	-	-	-	-	-	5	7	12
Non-Infected	-	-	NA	-	-	-	-	-	2	7	9
Non-Infected	-	Invalid	-	-	-	-	-	-	5	4	9
Non-Infected	-	-	-	-	-	Invalid	-	-	5	3	8
Non-Infected	-	-	-	-	-	-	-	NA	3	5	8
Non-Infected	-	-	Invalid	-	-	-	-	-	0	8	8
Non-Infected	-	-	-	-	-	+	-	-	2	4	6
Non-Infected	-	-	-	-	-	NA	-	-	1	3	4
Non-Infected	-	NA	-	-	-	-	-	-	0	4	4
Non-Infected	-	-	-	-	-	-	NA	NA	0	3	3
Non-Infected	-	-	-	NA	-	-	-	-	1	2	3
Non-Infected	NA	-	-	-	-	-	-	-	0	3	3
Non-Infected	Invalid	-	-	-	-	-	-	-	2	1	3
Non-Infected	+	+	-	-	-	+	-	-	0	2	2
Non-Infected	-	-	-	-	+	-	-	-	2	0	2
Non-Infected	-	-	-	-	-	-	-	Invalid	2	0	2

33

	NAAT1		NAAT2		cobas® CT/NG				Symptom Statusa
Patient
Infected
Status	UR	VS	UR	VS	UR	VS	PC	ES	Symp	Asymp	Total
Non-Infected	+	+	-	-	-	-	-	-	0	1	1
Non-Infected	+	+	-	-	-	Invalid	-	-	1	0	1
Non-Infected	-	+	-	-	-	+	+	-	1	0	1
Non-Infected	-	+	-	-	-	+	-	-	1	0	1
Non-Infected	-	-	+	+	+	-	-	-	1	0	1
Non-Infected	-	-	-	+	-	+	-	-	0	1	1
Non-Infected	-	-	-	-	-	-	+	+	0	1	1
Non-Infected	-	-	-	-	-	-	-	+	1	0	1
Non-Infected	-	-	-	-	-	Failed	-	-	1	0	1
Non-Infected	-	-	-	-	Failed	-	-	-	1	0	1
Non-Infected	-	-	-	-	-	-	-	Failed	1	0	1
Non-Infected	-	-	-	-	-	-	NA	-	0	1	1
Non-Infected	-	-	-	-	-	-	Invalid	-	1	0	1
Non-Infected	-	-	-	-	-	NA	Invalid	-	1	0	1
Non-Infected	-	-	-	-	-	-	Invalid	Invalid	1	0	1
Non-Infected	-	-	-	-	-	Invalid	Invalid	Invalid	1	0	1
Non-Infected	-	-	-	-	-	-	Failed	-	0	1	1
Non-Infected	-	-	-	Invalid	-	-	-	-	1	0	1
Non-Infected	-	Invalid	-	-	-	Invalid	-	-	1	0	1
Total									1416	2387	3803

ª Symp = symptomatic, Asymp = asymptomatic.

b Infected (Urine), Non-Infected (Swabs).

Note: In the scenario where one or more of the sample types are invalidinot available (NA), for female subjects, the remaining sample types with valid results from NAAT1 and NAAT2 must have concordant positive or concordant negative results to determine the PIS as Infected, respectively. For all other cases where one or more of the sample types are invalid/not available (NA), the PIS is indeterminate.

Note: Female subjects with designated infection status (Infected) and final valid cobas® CT/NG test results are considered evaluable and included in this summary table.

Note: cobas Invalid are the sum of instrument amplification errors and samples excluded due to protocol deviations

Note: cobas Failed are hardware, software or operator errors causing no result reported

Note: + denotes Positive, - denotes Negative, NA denotes Not Available.

Note: UR = urine, VS = vaginal swab, PC = PreservCyt®, ES = endocervical swab.

34

| | NAAT1a | NAAT2a | NAAT3a | cobas®
CT/NG | Symptom Statusb | | |
|-------------------------------|---------|---------|---------|-----------------|-----------------|-------|-------|
| Patient
Infected
Status | UR | UR | UR | UR | Symp | Asymp | Total |
| Infected | + | + | + | + | 81 | 5 | 86 |
| Infected | NA | + | + | + | 1 | 0 | 1 |
| Total Infected | | | | | 82 | 5 | 87 |
| Non-Infected | - | - | - | - | 215 | 863 | 1078 |
| Non-Infected | + | - | - | - | 2 | 7 | 9 |
| Non-Infected | - | Invalid | - | - | 3 | 2 | 5 |
| Non-Infected | - | - | - | + | 2 | 2 | 4 |
| Non-Infected | Invalid | - | - | - | 0 | 3 | 3 |
| Non-Infected | - | - | Invalid | - | 0 | 3 | 3 |
| Non-Infected | - | + | - | + | 1 | 1 | 2 |
| Non-Infected | NA | - | - | - | 0 | 1 | 1 |
| Total Non-
Infected | | | | | 223 | 882 | 1105* |

Table 17: NG positive/negative analysis for male Patient Infected Status

a Symp = symptomatic, Asymp = asymptomatic.

*One subject was missing symptom status and is not included in this table.

Note: If at least 2 out of the 3 test results, for male subjects, are concordant positive or negative then the PIS can be considered as infected or non-infected, respectively. If one test result is invalid/not available (NA) and the other two test results are discordant the PIS is indeterminate. If 2 or 3 test results are invalid/not available, then the PIS is indeterminate.

Note: Male subjects with designated patient infection status (Infected or Non-Infected) and final valid cobas® CT/NG test results are considered evaluable and included in this summary table.

Note: + denotes Positive, - denotes Negative, NA denotes Not Available.

Note: UR = urine.

Sensitivity, specificity, and predictive values of cobas® CT/NG for NG as defined by PIS are presented by gender, sample type, and symptom status in Table 18 (prospective fresh and archived specimens).

35

Table 18: NG clinical performance compared with Patient Infected Status by gender, sample type, and symptom status (prospective and archived prospectively collected specimens)

| Sample
Typea | Symptom
Statusb | Total
(n) | SENS | 95% Score
CI | SPEC | 95%
Score CI | PREV
(%) | PPV
(%) | NPV
(%) |
|-------------------------------------|--------------------|--------------|--------------------|--------------------|-----------------------|--------------------|-------------|------------|------------|
| Female | | | | | | | | | |
| UR
(prospective) | Symp | 1441 | 92.3%
(24/26) | (75.9%,
97.9%) | 99.8%
(1412/1415) | (99.4%,
99.9%) | 1.8 | 88.9 | 99.9 |
| | Asymp | 2418 | 87.1%
(27/31) | (71.1%,
94.9%) | 100.0%
(2387/2387) | (99.8%,
100.0%) | 1.3 | 100.0 | 99.8 |
| | Overall | 3859 | 89.5%
(51/57) c | (78.9%,
95.1%) | 99.9%
(3799/3802) | (99.8%,
100.0%) | 1.5 | 94.4 | 99.8 |
| UR
(archived) | Symp | 94 | 100.0%
(35/35) | (90.1%,
100.0%) | 100.0%
(59/59) | (93.9%,
100.0%) | 37.2 | 100.0 | 100.0 |
| | Asymp | 101 | 97.6%
(41/42) | (87.7%,
99.6%) | 100.0%
(59/59) | (93.9%,
100.0%) | 41.6 | 100.0 | 98.3 |
| | Overall | 195 | 98.7%
(76/77) | (93.0%,
99.8%) | 100.0%
(118/118) | (96.8%,
100.0%) | 39.5 | 100.0 | 99.2 |
| UR
(prospective
and archived) | Symp | 1535 | 96.7%
(59/61) | (88.8%,
99.1%) | 99.8%
(1471/1474) | (99.4%,
99.9%) | 4.0 | 95.2 | 99.9 |
| | Asymp | 2519 | 93.2%
(68/73) | (84.9%,
97.0%) | 100.0%
(2446/2446) | (99.8%,
100.0%) | 2.9 | 100.0 | 99.8 |
| | Overall | 4054 | 94.8%
(127/134) | (89.6%,
97.4%) | 99.9%
(3917/3920) | (99.8%,
100.0%) | 3.3 | 97.7 | 99.8 |
| VS-C | Symp | 711 | 100.0%
(11/11) | (74.1%,
100.0%) | 99.7%
(698/700) | (99.0%,
99.9%) | 1.5 | 84.6 | 100.0 |
| | Asymp | 1225 | 100.0%
(17/17) | (81.6%,
100.0%) | 99.8%
(1205/1208) | (99.3%,
99.9%) | 1.4 | 85.0 | 100.0 |
| | Overall | 1936 | 100.0%
(28/28) | (87.9%,
100.0%) | 99.7%
(1903/1908) | (99.4%,
99.9%) | 1.4 | 84.8 | 100.0 |
| VS-S | Symp | 720 | 100.0%
(14/14) | (78.5%,
100.0%) | 99.7%
(704/706) | (99.0%,
99.9%) | 1.9 | 87.5 | 100.0 |
| | Asymp | 1187 | 100.0%
(14/14) | (78.5%,
100.0%) | 99.7%
(1169/1173) | (99.1%,
99.9%) | 1.2 | 77.8 | 100.0 |
| | Overall | 1907 | 100.0%
(28/28) | (87.9%,
100.0%) | 99.7%
(1873/1879) | (99.3%,
99.9%) | 1.5 | 82.4 | 100.0 |
| PC
(prospective) | Symp | 1438 | 100.0%
(25/25) | (86.7%,
100.0%) | 99.9%
(1412/1413) | (99.6%,
100.0%) | 1.7 | 96.2 | 100.0 |
| | Asymp | 2413 | 93.5%
(29/31) | (79.3%,
98.2%) | 100.0%
(2381/2382) | (99.8%,
100.0%) | 1.3 | 96.7 | 99.9 |
| | Overall | 3851 | 96.4%
(54/56) | (87.9%,
99.0%) | 99.9%
(3793/3795) | (99.8%,
100.0%) | 1.5 | 96.4 | 99.9 |
| Sample
Typea | Symptom
Statusb | Total
(n) | SENS | 95% Score
CI | SPEC | 95%
Score Cl | PREV
(%) | PPV
(%) | NPV
(%) |
| PC
(archived) | Symp | 48 | 95.7%
(22/23) | (79.0%,
99.2%) | 100.0%
(25/25) | (86.7%,
100.0%) | 47.9 | 100.0 | 96.2 |
| | Asymp | 23 | 100.0%
(10/10) | (72.2%,
100.0%) | 100.0%
(13/13) | (77.2%,
100.0%) | 43.5 | 100.0 | 100.0 |
| | Overall | 71 | 97.0%
(32/33) | (84.7%,
99.5%) | 100.0%
(38/38) | (90.8%,
100.0%) | 46.5 | 100.0 | 97.4 |
| РС
(prospective | Symp | 1486 | 97.9%
(47/48) | (89.1%,
99.6%) | 99.9%
(1437/1438) | (99.6%,
100.0%) | 3.2 | 97.9 | 99.9 |
| and archived) | Asymp | 2436 | 95.1%
(39/41) | (83.9%,
98.7%) | 100.0%
(2394/2395) | (99.8%,
100.0%) | 1.7 | 97.5 | 99.9 |
| | Overall | 3922 | 96.6%
(86/89) | (90.6%,
98.8%) | 99.9%
(3831/3833) | (99.8%,
100.0%) | 2.3 | 97.7 | 99.9 |
| ES
(prospective) | Symp | 1433 | 100.0%
(24/24) | (86.2%,
100.0%) | 99.9%
(1408/1409) | (99.6%,
100.0%) | 1.7 | 96.0 | 100.0 |
| | Asymp | 2410 | 90.3%
(28/31) | (75.1%,
96.7%) | 100.0%
(2378/2379) | (99.8%,
100.0%) | 1.3 | 96.6 | 99.9 |
| | Overall | 3843 | 94.5%
(52/55) | (85.1%,
98.1%) | 99.9%
(3786/3788) | (99.8%,
100.0%) | 1.4 | 96.3 | 99.9 |
| ES
(archived) | Symp | 51 | 100.0%
(21/21) | (84.5%,
100.0%) | 100.0%
(30/30) | (88.6%,
100.0%) | 41.2 | 100.0 | 100.0 |
| | Asymp | 54 | 100.0%
(24/24) | (86.2%,
100.0%) | 100.0%
(30/30) | (88.6%,
100.0%) | 44.4 | 100.0 | 100.0 |
| | Overall | 105 | 100.0%
(45/45) | (92.1%,
100.0%) | 100.0%
(60/60) | (94.0%,
100.0%) | 42.9 | 100.0 | 100.0 |
| ES
(prospective | Symp | 1484 | 100.0%
(45/45) | (92.1%,
100.0%) | 99.9%
(1438/1439) | (99.6%,
100.0%) | 3.0 | 97.8 | 100.0 |
| and archived) | Asymp | 2464 | 94.5%
(52/55) | (85.1%,
98.1%) | 100.0%
(2408/2409) | (99.8%,
100.0%) | 2.2 | 98.1 | 99.9 |
| | Overall | 3948 | 97.0%
(97/100) | (91.5%,
99.0%) | 99.9%
(3846/3848) | (99.8%,
100.0%) | 2.5 | 98.0 | 99.9 |
| UR | Symp | 305 | 100.0%
(82/82) | (95.5%,
100.0%) | 98.7%
(220/223) | (96.1%,
99.5%) | 26.9 | 96.5 | 100.0 |
| | Asymp | 887 | 100.0%
(5/5) | (56.6%,
100.0%) | 99.7%
(879/882) | (99.0%,
99.9%) | 0.6 | 62.5 | 100.0 |
| | Overall | 1192* | 100.0%
(87/87) | (95.8%,
100.0%) | 99.5%
(1099/1105) | (98.8%,
99.8%) | 7.3 | 93.5 | 100.0 |

36

37

-----------------	--------------------	--------------	------	-----------------	------	-----------------	-------------	------------	------------

a UR = urine, VS-C = clinician-collected vaginal swab, VS-S = self-collected vaginal swab, PC = PreservCyt®, ES = endocervical swab.

b Symp = symptomatic. Asymp = asymptomatic.

^ Five NG PIS infected females had a NG negative urine specimen with NAAT1 and NAAT2 while they had a NG positive vaginal swab with NAAT1 and NAAT2

One subject was missing symptom status and is not included in this table.

Note: In the scenario where one or more of the sample types are invalid/not available, for female subjects, the remaining sample types with valid results from NAAT2 must have concordant positive or concordant negative results to determine the PIS as Infected, respectively. For all other cases where one or more of the sample types are invalid/ not available, the PIS is indeterminate.

Note: If at least 2 out of the 3 test results, for male subjects, are concordant positive or negative then the PIS can be considered as Infected or Non-Infected, respectively. If one test result is invalidinot available and the other two test results are discordant then the PIS is indeterminate. If 2 or 3 test results are invalid/not available, then the PIS is indeterminate.

Note: Subjects with designated patient infection status (Infected) and final valid cobas® CT/NG test results are considered evaluable and included in this summary table. An evaluable subject may not have all available sample types or valid test results.

Note: Archived prospectively collected specimens were from COB-CTNG-282 study and included female PIS positive subjects that have available sample with adequate volume for testing.

Note: Cl = confidence interval, PREV = prevalence, SENS = sensitivity, SPEC = specificity, PPV = positive predictive value, NPV = negative predictive value.

5.2. Clinical Reproducibility

Study Design 5.2.1.

A Reproducibility Study was performed across different sites, lots, operators/batches days, for cobas® CT/NG using three panels prepared from swabs and urine in cobas® PCR Media and cervical specimens in PreservCyt® Solution. PCR testing was performed at two external sites and one site that was in-house at Roche Molecular Systems. One panel consisted of the three sample matrices, with six concentrations per matrix, and three replicates per concentration for a total of 54 samples in one panel. A batch was comprised of one 54-sample panel and two controls (one positive control and one negative control). Two operators at each site tested one batch each per day. Two valid batches had to be completed within a 24-hour period. Each site received two of three reagent lots and performed 6 days of testing per reagent lot for a total of 12 days of testing.

38

5.2.2. Results

The Reproducibility Study was executed with a total of 3.888 tests performed on the 6 panel groups, consisting of 1.296 tests for each panel type (urine, swab, and PreservCyt®), with only two failed tests each from PreservCyt®. No false positive results for either CT or NG were observed in the three panel types for negative panel members; thus the negative percent agreement was 100% for each analyte. Results for the positive panel members were highly reproducible across different lots, sites/instruments, days and operators/batches.

5.2.2.1. Negative panel results

For each sample type, all of the 216 valid tests from the negative panel members resulted in "Negative Results". Hence, for both CT and NG, the percent of correct results (analytical specificity) was estimated as 100% with a corresponding 95% exact confidence interval of 98.3%, 100% for cobas® PCR Media/urine, for cobas® PCR Media/swab and for PreservCyt®/cervical sample types.

Chlamydia trachomatis results 5.2.2.2.

For each positive panel member, precision was evaluated using a random effects model by sample type with terms for lot, site, day, operator/batch within site, lot and day, and within-batch components on the corresponding analyte cycle threshold (Ct) values of cobas® CT/NG. Table 19 presents the total SD, and total percent CV (%) from these analyses for each panel type, respectively. The range of the total coefficient of variation, among positive panel members, was from 0.9% to 3.2%. The maximum total coefficient of variation was observed in the lowest concentration of positive panel members (0.3x LoD CT, 0.3x LoD NG) and most of that variability (98.6% for urine, 100% for swab and 81.7% for cervical) was explained by random error (within-batch).

39

Table 19: CT: overall mean, attributable percentage of total variance, total precision standard deviation, and CV(%) of cobas® CT/NG cycle threshold (Ct) values by CT positive panel member for each media type

Panel Member	Mean CT value		Percentage of Total Variance (CV[%])					Total Precision
Media Type	Concentration	N a	Mean Estimate b	Site	Lot	Day	Operator /Batch	Within-Batch	SD c	CV(%) d
PCR Media/Urine	0.3xLOD CT, 0.3xLOD NG	154	39.2	1.4% (0.4)	0.0% (0.0)	0.0% (0.0)	0.0% (0.0)	98.6% (3.0)	1.20	3.1
	1xLOD CT, Negative NG	216	36.8	0.0% (0.0)	0.0% (0.0)	0.0% (0.0)	0.0% (0.0)	100.0% (1.5)	0.54	1.5
	3xLOD CT, 1xLOD NG	216	35.4	2.4% (0.1)	0.0% (0.0)	21.1% (0.4)	0.0% (0.0)	76.5% (0.8)	0.33	0.9
	1xLOD CT, 3xLOD NG	216	36.9	0.0% (0.0)	0.0% (0.0)	10.3% (0.5)	4.4% (0.3)	85.3% (1.5)	0.59	1.6
PCR Media/Swab	0.3xLOD CT, 0.3xLOD NG	128	39.5	0.0% (0.0)	0.0% (0.0)	0.0% (0.0)	0.0% (0.0)	100.0% (3.2)	1.26	3.2
	1xLOD CT, Negative NG	216	37.2	0.0% (0.0)	1.6% (0.2)	6.6% (0.5)	0.0% (0.0)	91.8% (1.7)	0.66	1.8
	3xLOD CT, 1xLOD NG	216	35.5	4.7% (0.2)	0.0% (0.0)	9.0% (0.3)	4.8% (0.2)	81.6% (0.9)	0.37	1.0
	1xLOD CT, 3xLOD NG	216	37.2	0.0% (0.0)	0.0% (0.0)	3.6% (0.4)	0.0% (0.0)	96.4% (2.3)	0.87	2.3
PreservCyt/Cervical	0.3xLOD CT, 0.3xLOD NG	92	39.9	0.0% (0.0)	0.0% (0.0)	18.3% (1.4)	0.0% (0.0)	81.7% (2.9)	1.29	3.2
	1xLOD CT, Negative NG	216	37.0	12.0% (0.6)	1.9% (0.2)	0.0% (0.0)	0.0% (0.0)	86.2% (1.5)	0.60	1.6
	3xLOD CT, 1xLOD NG	216	35.6	0.6% (0.1)	3.7% (0.2)	0.0% (0.0)	6.3% (0.3)	89.3% (0.9)	0.36	1.0
	1xLOD CT, 3xLOD NG	214	36.8	13.1% (0.6)	3.7% (0.3)	5.3% (0.4)	2.3% (0.3)	75.6% (1.5)	0.63	1.7

Note: The table only includes results with detectable analyte. SD = standard deviation. CV(%) = percent coefficient of variation.

ª Number of valid tests with detectable analyte.

b Calculated using SAS MIXED procedure.

َ Calculated using the total variability from the SAS MIXED procedure.

d CV(%) = (SD/Mean) * 100.

LOD= Limit of Detection; CT= Chlamydia trachomatis ; NG = Neisseria gonorrhoeae.

40

Table 20 through Table 22 present the percent agreement of CT test results for panel members by lot, site, and day for each media type, respectively.

				CT Percent Agreementa
Panel
Member	Ct
SD	Ct
CV
%	Lot		Siteb		Day
Negative CT, Negative
NG	n/a	n/a	1	100.0	72/72	1	100.0	72/72	1	100.0	36/36
			2	100.0	72/72	2	100.0	72/72	2	100.0	36/36
			3	100.0	72/72	3	100.0	72/72	3	100.0	36/36
									4	100.0	36/36
									5	100.0	36/36
									6	100.0	36/36
0.3x LoD CT, 0.3x LoD
NG	1.20	3.1	1	76.4	55/72	1	68.1	49/72	1	80.6	29/36
			2	70.8	51/72	2	73.6	53/72	2	77.8	28/36
			3	66.7	48/72	3	72.2	52/72	3	66.7	24/36
									4	77.8	28/36
									5	69.4	25/36
									6	55.6	20/36
1x LoD CT, Negative NG	0.54	1.5	1	100.0	72/72	1	100.0	72/72	1	100.0	36/36
			2	100.0	72/72	2	100.0	72/72	2	100.0	36/36
			3	100.0	72/72	3	100.0	72/72	3	100.0	36/36
									4	100.0	36/36
									5	100.0	36/36
									6	100.0	36/36
Negative CT, 1x LoD NG	n/a	n/a	1	100.0	72/72	1	100.0	72/72	1	100.0	36/36
			2	100.0	72/72	2	100.0	72/72	2	100.0	36/36
			3	100.0	72/72	3	100.0	72/72	3	100.0	36/36
									4	100.0	36/36
									5	100.0	36/36
									6	100.0	36/36
Panel
Member	Ct
SD	Ct
CV
%	Lot			Siteb			Day
3x LoD CT, 1x LoD NG	0.33	0.9	1	100.0	72/72	1	100.0	72/72	1	100.0	36/36
			2	100.0	72/72	2	100.0	72/72	2	100.0	36/36
			3	100.0	72/72	3	100.0	72/72	3	100.0	36/36
									4	100.0	36/36
									5	100.0	36/36
									6	100.0	36/36
1x LoD CT, 3x LoD NG	0.59	1.6	1	100.0	72/72	1	100.0	72/72	1	100.0	36/36
			2	100.0	72/72	2	100.0	72/72	2	100.0	36/36
			3	100.0	72/72	3	100.0	72/72	3	100.0	36/36
									4	100.0	36/36
									5	100.0	36/36
									6	100.0	36/36

Table 20: CT: Percent agreement by panel member for lot, site and day - cobas® PCR Media/urine

41

ª For CT Negative samples, Percent Agreement = (number of CT negative results/total valid results) x 100. For CT Positive samples, Percent Agreement = (number of CT positive results/total valid results) x 100. b Site 1, Site 2, and Site 3, respectively.

Ct=Cycle threshold; SD=Standard Deviation; CV=Coefficient of Variation; LoD= Limit of Detection.

CT= Chlamydia trachomatis; NG = Neisseria gonorrhoeae; n/a= not applicable.

Table 21: CT: Percent agreement by panel member for lot, site and day - cobas® PCR Media/swab

			CT Percent Agreementa
Panel
Member	Ct
SD	Ct
CV
%	Lot		Siteb		Day
Negative CT, Negative
NG	n/a	n/a	1	100.0	72/72	1	100.0	72/72	1	100.0	36/36
			2	100.0	72/72	2	100.0	72/72	2	100.0	36/36
			3	100.0	72/72	3	100.0	72/72	3	100.0	36/36
									4	100.0	36/36
									5	100.0	36/36
									6	100.0	36/36
0.3x LoD CT, 0.3x LoD
NG	1.26	3.2	1	61.1	44/72	1	56.9	41/72	1	50.0	18/36
			2	59.7	43/72	2	61.1	44/72	2	63.9	23/36
			3	56.9	41/72	3	59.7	43/72	3	55.6	20/36
									4	61.1	22/36
									5	66.7	24/36
									6	58.3	21/36

42

| Panel
Member | Ct
SD | Ct
CV
% | Lot | | | | Siteb | | Day | | |
|------------------------|----------|---------------|-----|-------|-------|---|-------|-------|-----|-------|-------|
| 1x LoD CT, Negative NG | 0.66 | 1.8 | 1 | 100.0 | 72/72 | 1 | 100.0 | 72/72 | 1 | 100.0 | 36/36 |
| | | | 2 | 100.0 | 72/72 | 2 | 100.0 | 72/72 | 2 | 100.0 | 36/36 |
| | | | 3 | 100.0 | 72/72 | 3 | 100.0 | 72/72 | 3 | 100.0 | 36/36 |
| | | | | | | | | | 4 | 100.0 | 36/36 |
| | | | | | | | | | 5 | 100.0 | 36/36 |
| | | | | | | | | | 6 | 100.0 | 36/36 |
| Negative CT, 1x LoD NG | n/a | n/a | 1 | 100.0 | 72/72 | 1 | 100.0 | 72/72 | 1 | 100.0 | 36/36 |
| | | | 2 | 100.0 | 72/72 | 2 | 100.0 | 72/72 | 2 | 100.0 | 36/36 |
| | | | 3 | 100.0 | 72/72 | 3 | 100.0 | 72/72 | 3 | 100.0 | 36/36 |
| | | | | | | | | | 4 | 100.0 | 36/36 |
| | | | | | | | | | 5 | 100.0 | 36/36 |
| | | | | | | | | | 6 | 100.0 | 36/36 |
| 3x LoD CT, 1x LoD NG | 0.37 | 1.0 | 1 | 100.0 | 72/72 | 1 | 100.0 | 72/72 | 1 | 100.0 | 36/36 |
| | | | 2 | 100.0 | 72/72 | 2 | 100.0 | 72/72 | 2 | 100.0 | 36/36 |
| | | | 3 | 100.0 | 72/72 | 3 | 100.0 | 72/72 | 3 | 100.0 | 36/36 |
| | | | | | | | | | 4 | 100.0 | 36/36 |
| | | | | | | | | | 5 | 100.0 | 36/36 |
| | | | | | | | | | 6 | 100.0 | 36/36 |
| 1x LoD CT, 3x LoD NG | 0.87 | 2.3 | 1 | 100.0 | 72/72 | 1 | 100.0 | 72/72 | 1 | 100.0 | 36/36 |
| | | | 2 | 100.0 | 72/72 | 2 | 100.0 | 72/72 | 2 | 100.0 | 36/36 |
| | | | 3 | 100.0 | 72/72 | 3 | 100.0 | 72/72 | 3 | 100.0 | 36/36 |
| | | | | | | | | | 4 | 100.0 | 36/36 |
| | | | | | | | | | 5 | 100.0 | 36/36 |
| | | | | | | | | | 6 | 100.0 | 36/36 |

ª For CT Negative samples, Percent Agreement = (number of CT negative results/total valid results) x 100.

For CT Positive samples, Percent Agreement = (number of CT positive results/total valid results) x 100.

b Site 1, Site 2, and Site 3, respectively.

Ct=Cycle threshold; SD=Standard Deviation; CV=Coefficient of Variation; LoD= Limit of Detection.

CT= Chlamydia trachomatis; NG = Neisseria gonorrhoeae; n/a= not applicable.

Table 22: CT: Percent agreement by panel member for lot, site and day -PreservCyt®/cervical

	CT Percent Agreementa
--	-----------------------

43

| Panel
Member | Ct
SD | Ct
CV
% | Lot | | Siteb | | Day | | | | |
|-----------------------------|----------|---------------|-----|-------|-------|---|-------|-------|---|-------|-------|
| Negative CT, Negative NG | n/a | n/a | 1 | 100.0 | 72/72 | 1 | 100.0 | 72/72 | 1 | 100.0 | 36/36 |
| | | | 2 | 100.0 | 72/72 | 2 | 100.0 | 72/72 | 2 | 100.0 | 36/36 |
| | | | 3 | 100.0 | 72/72 | 3 | 100.0 | 72/72 | 3 | 100.0 | 36/36 |
| | | | | | | | | | 4 | 100.0 | 36/36 |
| | | | | | | | | | 5 | 100.0 | 36/36 |
| | | | | | | | | | 6 | 100.0 | 36/36 |
| 0.3x LoD CT, 0.3x LoD
NG | 1.29 | 3.2 | 1 | 38.9 | 28/72 | 1 | 34.7 | 25/72 | 1 | 40.0 | 14/35 |
| | | | 2 | 47.9 | 34/71 | 2 | 48.6 | 35/72 | 2 | 52.8 | 19/36 |
| | | | 3 | 41.7 | 30/72 | 3 | 45.1 | 32/71 | 3 | 38.9 | 14/36 |
| | | | | | | | | | 4 | 47.2 | 17/36 |
| | | | | | | | | | 5 | 41.7 | 15/36 |
| | | | | | | | | | 6 | 36.1 | 13/36 |
| 1x LoD CT, Negative NG | 0.60 | 1.6 | 1 | 100.0 | 72/72 | 1 | 100.0 | 72/72 | 1 | 100.0 | 36/36 |
| | | | 2 | 100.0 | 72/72 | 2 | 100.0 | 72/72 | 2 | 100.0 | 36/36 |
| | | | 3 | 100.0 | 72/72 | 3 | 100.0 | 72/72 | 3 | 100.0 | 36/36 |
| | | | | | | | | | 4 | 100.0 | 36/36 |
| | | | | | | | | | 5 | 100.0 | 36/36 |
| | | | | | | | | | 6 | 100.0 | 36/36 |
| Negative CT, 1x LoD NG | n/a | n/a | 1 | 100.0 | 72/72 | 1 | 100.0 | 72/72 | 1 | 100.0 | 36/36 |
| | | | 2 | 100.0 | 72/72 | 2 | 100.0 | 72/72 | 2 | 100.0 | 36/36 |
| | | | 3 | 100.0 | 72/72 | 3 | 100.0 | 72/72 | 3 | 100.0 | 36/36 |
| | | | | | | | | | 4 | 100.0 | 36/36 |
| | | | | | | | | | 5 | 100.0 | 36/36 |
| | | | | | | | | | 6 | 100.0 | 36/36 |
| 3x LoD CT, 1x LoD NG | 0.36 | 1.0 | 1 | 100.0 | 72/72 | 1 | 100.0 | 72/72 | 1 | 100.0 | 36/36 |
| | | | 2 | 100.0 | 72/72 | 2 | 100.0 | 72/72 | 2 | 100.0 | 36/36 |
| | | | 3 | 100.0 | 72/72 | 3 | 100.0 | 72/72 | 3 | 100.0 | 36/36 |
| | | | | | | | | | 4 | 100.0 | 36/36 |
| | | | | | | | | | 5 | 100.0 | 36/36 |
| | | | | | | | | | 6 | 100.0 | 36/36 |
| 1x LoD CT, 3x LoD NG | 0.63 | 1.7 | 1 | 98.6 | 71/72 | 1 | 98.6 | 71/72 | 1 | 97.2 | 35/36 |
| | | | 2 | 100.0 | 71/71 | 2 | 100.0 | 71/71 | 2 | 100.0 | 36/36 |
| | | | 3 | 100.0 | 72/72 | 3 | 100.0 | 72/72 | 3 | 100.0 | 36/36 |

44

	CT Percent Agreementa
Panel
Member	Ct
SD	Ct
CV
%	Lot	Siteb	Day
				4	100.0	35/35
				5	100.0	36/36
				6	100.0	36/36

a For CT Negative samples, Percent Agreement = (number of CT negative results/total valid results) x 100. For CT Positive samples, Percent Agreement = (number of CT positive results/total valid results) x 100. b Site 1, Site 2, and Site 3, respectively.

Ct=Cycle threshold; SD=Standard Deviation; CV=Coefficient of Variation; LoD= Limit of Detection. CT= Chlamydia trachomatis; NG = Neisseria gonorrhoeae; n/a= not applicable.

Neisseria gonorrhoeae results 5.2.2.3.

Analysis of variance components of the Ct values from valid NG test results were performed on positive panel members. Table 23 presents the total CV (%) from these analyses. The range of the total coefficient of variation, among positive panel members, was from 1.0% to 3.1%. The maximum total coefficient of variation was observed in the lowest concentration of positive panel members (0.3x LoD CT, 0.3x LoD NG) and most of that variability (98.7% for urine, 98.1% for swab and 85.3% for cervical) was explained by random error (within-batch).

45

Table 23: NG: overall mean, attributable percentage of total variance, total precision standard deviation, and CV(%) of cobas® CTNG cycle threshold (Ct) values by NG positive panel member for each media type

Panel Member	Mean CT value		Percentage of Total Variance (CV[%])		Total Precision
Media Type	Concentration	N a	Mean Estimate b	Site	Lot	Day	Operator/Batch	Within-Batch	SD c	CV(%) d
PCR Media/Urine	0.3xLOD CT, 0.3xLOD NG	159	39.3	0.7%
(0.3)	0.0%
(0.0)	0.6%
(0.2)	0.0%
(0.0)	98.7%
(3.0)	1.20	3.0
	Negative CT, 1xLOD NG	216	36.7	0.0%
(0.0)	0.5%
(0.1)	6.9%
(0.5)	0.0%
(0.0)	92.6%
(1.7)	0.63	1.7
	3xLOD CT, 1xLOD NG	216	36.6	0.0%
(0.0)	2.5%
(0.3)	8.3%
(0.5)	0.0%
(0.0)	89.2%
(1.6)	0.61	1.7
	1xLOD CT, 3xLOD NG	216	35.1	0.0%
(0.0)	0.0%
(0.0)	14.0%
(0.4)	0.0%
(0.0)	86.0%
(1.0)	0.37	1.0
PCR Media/Swab	0.3xLOD CT, 0.3xLOD NG	113	39.8	0.0%
(0.0)	0.0%
(0.0)	1.9%
(0.4)	0.0%
(0.0)	98.1%
(3.1)	1.25	3.1
	Negative CT, 1xLOD NG	212	38.2	0.0%
(0.0)	0.1%
(0.1)	1.8%
(0.4)	6.5%
(0.7)	91.6%
(2.6)	1.04	2.7
	3xLOD CT, 1xLOD NG	216	36.9	0.0%
(0.0)	0.0%
(0.0)	6.3%
(0.6)	0.0%
(0.0)	93.7%
(2.1)	0.82	2.2
	1xLOD CT, 3xLOD NG	216	35.7	0.0%
(0.0)	3.8%
(0.3)	14.4%
(0.5)	0.0%
(0.0)	81.8%
(1.3)	0.50	1.4
PreservCyt/Cervical	0.3xLOD CT, 0.3xLOD NG	112	39.5	0.0%
(0.0)	0.0%
(0.0)	0.0%
(0.0)	14.7%
(1.0)	85.3%
(2.4)	1.04	2.6
	Negative CT, 1xLOD NG	216	35.7	7.2%
(0.4)	4.9%
(0.3)	0.0%
(0.0)	0.0%
(0.0)	87.9%
(1.3)	0.49	1.4
	3xLOD CT, 1xLOD NG	216	36.3	0.0%
(0.0)	0.0%
(0.0)	0.0%
(0.0)	9.6%
(0.5)	90.4%
(1.6)	0.61	1.7
	1xLOD CT, 3xLOD NG	215	34.6	2.3%
(0.2)	0.0%
(0.0)	5.8%
(0.2)	12.0%
(0.3)	79.8%
(0.9)	0.34	1.0
Note: The table only includes results with detectable analyte. SD = standard deviation. CV(%) = percent coefficient of variation.

a Number of valid tests with detectable analyte.

b Calculated using SAS MIXED procedure.

· Calculated using the total variability from the SAS MIXED procedure.

d CV(%) = (SD/Mean) * 100.

LOD= Limit of Detection; CT = Chlamydia trachomatis ; NG = Neisseria gonorrhoeae.

Table 24 through Table 26 present the percent agreement of NG test results for panel members

by lot, site, and day for each media type, respectively.

46

		NG Percent Agreementa
Panel
Member	Ct
SD	Ct
CV
%	Lot			Siteb	Day
Negative CT, Negative NG	n/a	n/a	1	100.0	72/72	100.0	1	100.0	36/36
			2	100.0	72/72	100.0	2	100.0	36/36
			3	100.0	72/72	100.0	3	100.0	36/36
							4	100.0	36/36
							5	100.0	36/36
							6	100.0	36/36
0.3x LoD CT, 0.3x LoD
NG	1.20	3.0	1	79.2	57/72	70.8	1	77.8	28/36
			2	73.6	53/72	76.4	2	75.0	27/36
			3	68.1	49/72	73.6	3	72.2	26/36
							4	80.6	29/36
							5	61.1	22/36
							6	75.0	27/36
1x LoD CT, Negative NG	n/a	n/a	1	100.0	72/72	100.0	1	100.0	36/36
			2	100.0	72/72	100.0	2	100.0	36/36
			3	100.0	72/72	100.0	3	100.0	36/36
							4	100.0	36/36
							5	100.0	36/36
							6	100.0	36/36
Negative CT, 1x LoD NG	0.63	1.7	1	100.0	72/72	100.0	1	100.0	36/36
			2	100.0	72/72	100.0	2	100.0	36/36
			3	100.0	72/72	100.0	3	100.0	36/36
							4	100.0	36/36
							5	100.0	36/36
							6	100.0	36/36
3x LoD CT, 1x LoD NG	0.61	1.7	1	100.0	72/72	100.0	1	100.0	36/36
			2	100.0	72/72	100.0	2	100.0	36/36
			3	100.0	72/72	100.0	3	100.0	36/36
							4	100.0	36/36
							5	100.0	36/36
							6	100.0	36/36

Table 24: NG: Percent agreement by panel member for lot, site and day - cobas® PCR Media/urine

47

| Panel
Member | Ct
SD | Ct
CV
% | NG Percent Agreementa
Lot | | | Siteb | | | Day | | |
|----------------------|----------|---------------|------------------------------|-------|-------|-------|-------|-------|-----|-------|-------|
| | | | | | | | | | | | |
| 1x LoD CT, 3x LoD NG | 0.37 | 1.0 | 1 | 100.0 | 72/72 | 1 | 100.0 | 72/72 | 1 | 100.0 | 36/36 |
| | | | 2 | 100.0 | 72/72 | 2 | 100.0 | 72/72 | 2 | 100.0 | 36/36 |
| | | | 3 | 100.0 | 72/72 | 3 | 100.0 | 72/72 | 3 | 100.0 | 36/36 |
| | | | | | | | | | 4 | 100.0 | 36/36 |
| | | | | | | | | | 5 | 100.0 | 36/36 |
| | | | | | | | | | 6 | 100.0 | 36/36 |

ª For NG Negative samples, Percent Agreement = (number of NG negative results/total valid results) x 100.

For NG Positive samples, Percent Agreement = (number of NG positive results/total valid results) x 100. b Site 1, Site 2, and Site 3, respectively.

Ct=Cycle threshold; SD=Standard Deviation; CV=Coefficient of Variation; LoD= Limit of Detection.

CT= Chlamydia trachomatis; NG = Neisseria gonorrhoeae; n/a= not applicable.

Table 25: NG: Percent agreement by panel member for lot, site and day - cobas® PCR
Media/swab

	NG Percent Agreementa
Panel
Member	Ct
SD	Ct
CV
%	Lot	Siteb	Day
Negative CT, Negative
NG	n/a	n/a	1 100.0 72/72	1 100.0 72/72	1 100.0 36/36
			2 100.0 72/72	2 100.0 72/72	2 100.0 36/36
			3 100.0 72/72	3 100.0 72/72	3 100.0 36/36
				4 100.0 36/36
				5 100.0 36/36
				6 100.0 36/36
0.3x LoD CT, 0.3x LoD
NG	1.25	3.1	1 50.0 36/72	1 50.0 36/72	1 52.8 19/36
			2 51.4 37/72	2 52.8 38/72	2 55.6 20/36
			3 55.6 40/72	3 54.2 39/72	3 44.4 16/36
				4 55.6 20/36
				5 52.8 19/36
				6 52.8 19/36

48

	NG Percent Agreementa
Panel
Member	Ct
SD	Ct
CV
%	Lot		Siteb		Day
1x LoD CT, Negative NG	n/a	n/a	1	100.0 72/72	1	100.0 72/72	100.0 36/36
			2	100.0 72/72	2	100.0 72/72	100.0 36/36
			3	100.0 72/72	3	100.0 72/72	100.0 36/36
							100.0 36/36
							100.0 36/36
							100.0 36/36
Negative CT, 1x LoD NG	1.04	2.7	1	100.0 72/72	1	97.2 70/72	100.0 36/36
			2	100.0 72/72	2	100.0 72/72	100.0 36/36
			3	94.4 68/72	3	97.2 70/72	97.2 35/36
							100.0 36/36
							97.2 35/36
							94.4 34/36
3x LoD CT, 1x LoD NG	0.82	2.2	1	100.0 72/72	1	100.0 72/72	100.0 36/36
			2	100.0 72/72	2	100.0 72/72	100.0 36/36
			3	100.0 72/72	3	100.0 72/72	100.0 36/36
							100.0 36/36
							100.0 36/36
							100.0 36/36
1x LoD CT, 3x LoD NG	0.50	1.4	1	100.0 72/72	1	100.0 72/72	100.0 36/36
			2	100.0 72/72	2	100.0 72/72	100.0 36/36
			3	100.0 72/72	3	100.0 72/72	100.0 36/36
							100.0 36/36
							100.0 36/36
							100.0 36/36

ª For NG Negative samples, Percent Agreement = (number of NG negative results/total valid results) x 100.

For NG Positive samples, Percent Agreement = (number of NG positive results/total valid results) x 100. b Site 1, Site 2, and Site 3, respectively.

Ct=Cycle threshold; SD=Standard Deviation; CV=Coefficient of Variation; LoD= Limit of Detection.

CT= Chlamydia trachomatis; NG = Neisseria gonorrhoeae; n/a= not applicable.

49

| Panel
Member | Ct
SD | Ct
CV
% | NG Percent Agreementa | | |
|--------------------------|----------|---------------|-----------------------|---------------|---------------|
| | | | Lot | Siteb | Day |
| Negative CT, Negative NG | n/a | n/a | 1 100.0 72/72 | 1 100.0 72/72 | 1 100.0 36/36 |
| | | | 2 100.0 72/72 | 2 100.0 72/72 | 2 100.0 36/36 |
| | | | 3 100.0 72/72 | 3 100.0 72/72 | 3 100.0 36/36 |
| | | | | | 4 100.0 36/36 |
| | | | | | 5 100.0 36/36 |
| | | | | | 6 100.0 36/36 |
| 0.3x LoD CT, 0.3x LoD NG | 1.04 | 2.6 | 1 63.9 46/72 | 1 59.7 43/72 | 1 54.3 19/35 |
| | | | 2 47.9 34/71 | 2 52.8 38/72 | 2 55.6 20/36 |
| | | | 3 44.4 32/72 | 3 43.7 31/71 | 3 47.2 17/36 |
| | | | | | 4 55.6 20/36 |
| | | | | | 5 52.8 19/36 |
| | | | | | 6 47.2 17/36 |
| 1x LoD CT, Negative NG | n/a | n/a | 1 100.0 72/72 | 1 100.0 72/72 | 1 100.0 36/36 |
| | | | 2 100.0 72/72 | 2 100.0 72/72 | 2 100.0 36/36 |
| | | | 3 100.0 72/72 | 3 100.0 72/72 | 3 100.0 36/36 |
| | | | | | 4 100.0 36/36 |
| | | | | | 5 100.0 36/36 |
| | | | | | 6 100.0 36/36 |
| Negative CT, 1x LoD NG | 0.49 | 1.4 | 1 100.0 72/72 | 1 100.0 72/72 | 1 100.0 36/36 |
| | | | 2 100.0 72/72 | 2 100.0 72/72 | 2 100.0 36/36 |
| | | | 3 100.0 72/72 | 3 100.0 72/72 | 3 100.0 36/36 |
| | | | | | 4 100.0 36/36 |
| | | | | | 5 100.0 36/36 |
| | | | | | 6 100.0 36/36 |
| 3x LoD CT, 1x LoD NG | 0.61 | 1.7 | 1 100.0 72/72 | 1 100.0 72/72 | 1 100.0 36/36 |
| | | | 2 100.0 72/72 | 2 100.0 72/72 | 2 100.0 36/36 |
| | | | 3 100.0 72/72 | 3 100.0 72/72 | 3 100.0 36/36 |
| | | | | | 4 100.0 36/36 |
| | | | | | 5 100.0 36/36 |
| | | | | | 6 100.0 36/36 |

Table 26: NG: Percent agreement by panel member for lot, site and day -PreservCyt®/cervical

50

		NG Percent Agreementa
Panel
Member	Ct
SD	Ct
CV
%	Lot			Siteb			Day

1x LoD CT, 3x LoD NG	0.34	1.0	1	100.0	72/72	1	100.0	72/72	1	100.0	36/36
			2	100.0	71/71	2	100.0	71/71	2	100.0	36/36
			3	100.0	72/72	3	100.0	72/72	3	100.0	36/36
									4	100.0	35/35
									5	100.0	36/36
									6	100.0	36/36

ª For NG Negative samples, Percent Agreement = (number of NG negative results/total valid results) x 100. For NG Positive samples, Percent Agreement = (number of NG positive results/total valid results) x 100.

b Site 1, Site 2, and Site 3, respectively.

Ct=Cycle threshold; SD=Standard Deviation; CV=Coefficient of Variation; LoD= Limit of Detection. CT= Chlamydia trachomatis; NG = Neisseria gonorrhoeae; n/a= not applicable.

Percentage agreement results 5.2.2.4.

Table 27 shows the percent agreement for each target (CT, NG) with the associated

95% Exact CI.

51

		CT		NG
Media Type	Panel
Member	Percent
Agreement	Percent
Agreement
95% Exact Cl	Percent
Agreement	Percent
Agreement
95% Exact Cl
PCR
Media/Urine	1.0x LoD CT,
Negative NG	100.0 (216/216)	(98.3, 100.0)	100.0 (216/216)	(98.3, 100.0)
	Negative CT,
1.0x LoD NG	100.0 (216/216)	(98.3, 100.0)	100.0 (216/216)	(98.3, 100.0)
	3.0x LoD CT,
1.0x LoD NG	100.0 (216/216)	(98.3, 100.0)	100.0 (216/216)	(98.3, 100.0)
	1.0x LoD CT,
3.0x LoD NG	100.0 (216/216)	(98.3, 100.0)	100.0 (216/216)	(98.3, 100.0)
PCR
Media/Swab	1.0x LoD CT,
Negative NG	100.0 (216/216)	(98.3, 100.0)	100.0 (216/216)	(98.3, 100.0)
	Negative CT,
1.0x LoD NG	100.0 (216/216)	(98.3, 100.0)	98.1 (212/216)	(95.3, 99.5)
	3.0x LoD CT,
1.0x LoD NG	100.0 (216/216)	(98.3, 100.0)	100.0 (216/216)	(98.3, 100.0)
	1.0x LoD CT,
3.0x LoD NG	100.0 (216/216)	(98.3, 100.0)	100.0 (216/216)	(98.3, 100.0)
PreservCyt®/
Cervical	1.0x LoD CT,
Negative NG	100.0 (216/216)	(98.3, 100.0)	100.0 (216/216)	(98.3, 100.0)
	Negative CT,
1.0x LoD NG	100.0 (216/216)	(98.3, 100.0)	100.0 (216/216)	(98.3, 100.0)
	3.0x LoD CT,
1.0x LoD NG	100.0 (216/216)	(98.3, 100.0)	100.0 (216/216)	(98.3, 100.0)
	1.0x LoD CT,
3.0x LoD NG	99.5 (214/215)	(97.4, 100.0)	100.0 (215/215)	(98.3, 100.0)

Table 27: Percent agreement for panel members with concentration at or near the LoD (1x LoD) or 3x LoD

Notes: LoD= Limit of Detection; CT= Chlamydia trachomatis; NG = Neisseria gonorrhoeae. For panel members with concentrations at or near the limit of detection (e.g., 1x LoD) of the test, the lower limit of the 2-sided 95% exact Cl of the percentage of correct test results should be equal to or greater than 91%. For panel members with concentrations 3-times above the limit of detection (e.g., 3x LoD) of the test, the lower limit of the 2-sided 95% exact Cl of the percentage of correct test results should be equal to or greater than 98%.

For panel members with concentrations at or near the limit of detection (e.g., 1x LoD) of the test, the lower limit of the 2-sided 95% exact CI of the percentage of correct test results was at least 97.4% for CT and 95.3% for NG.

52

For panel members with concentrations 3-times above the limit of detection (e.g., 3x LoD) of the test, the lower limit of the 2-sided 95% exact CI of the percentage of correct test results was 98.3% for both CT and NG.

CONCLUSIONS 6.

A comparison of the intended use, technological characteristics, and the results of non-clinical analytical and clinical performance studies demonstrate that cobas® CT/NG for use on the cobas® 6800/8800 systems is substantially equivalent to the predicate device.