Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The summary describes a standard nucleic acid amplification test (NAAT) performed on an automated system. There is no mention of AI, ML, or any computational methods beyond basic signal processing for detection.

Therapeutic

No
The device is an in vitro diagnostic assay intended to aid in the diagnosis of vulvovaginal candidiasis and trichomoniasis by detecting specific RNA from microorganisms, not to treat the conditions.

Diagnostic

Yes

The "Intended Use / Indications for Use" section explicitly states that the assay is "intended to aid in the diagnosis of vulvovaginal candidiasis and trichomoniasis".

SaMD (Software as a Medical Device)

No

The device is an in vitro diagnostic (IVD) assay that utilizes a hardware system (Panther system) for nucleic acid amplification and detection. It is not solely software.

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 assay is "in vitro nucleic acid amplification" and is "intended to aid in the diagnosis of vulvovaginal candidiasis and trichomoniasis". This clearly indicates that the device is used outside of the body to analyze biological samples for diagnostic purposes.
Device Description: The "Device Description" further reinforces this by describing the assay as an "in vitro nucleic acid amplification test" that provides "qualitative results to aid in the diagnosis".
Sample Type: The assay uses "clinician-collected and patient-collected vaginal swab specimens", which are biological samples.
Purpose: The purpose is to detect and identify specific microorganisms (Candida species group, Candida glabrata, and Trichomonas vaginalis) associated with specific medical conditions (vulvovaginal candidiasis and trichomoniasis) to aid in diagnosis.

All of these points align with the definition of an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The Aptima CV/TV assay is an in vitro nucleic acid amplification of RNA from microorganisms associated with vulvovaginal candidiasis and trichomoniasis. The assay utilizes real time transcription-mediated amplification (TMA) to detect and qualitatively report results for the following organisms:

Candida species group (C. albicans, C. tropicalis, C. parapsilosis, C. dubliniensis)
Candida glabrata
Trichomonas vaginalis

The assay differentiates between Candida glabrata and the Candida species group (C spp) by targeting the RNA component of RNAse Pribonucleoprotein; the assay does not differentiate among C spp. For Trichomonas vaginalis, the assay targets ribosomal RNA (rRNA) and differentiates the results for Candida glabrata and C spp. The assay is intended to aid in the diagnosis of vulvovaginal candidiasis and trichomoniasis on the automated Panther system using clinician-collected and patient-collected vaginal swab specimens from females with a clinical presentation consistent with vaginitis or vulvovaginitis.

Product codes

PQA, NSU

Device Description

The Aptima CV/TV assay is an in vitro nucleic acid amplification test for the detection and quantitation of RNA from microorganisms associated with vulvovaginal candidiasis and trichomoniasis in women with a clinical presentation consistent with vaginitis or vulvovaginitis. The Aptima CV/TV assay utilizes the automated Panther system to provide qualitative results to aid in the diagnosis of vulvovaginal candidiasis and trichomoniasis.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

vagina

Indicated Patient Age Range

≥14 years

Intended User / Care Setting

Clinician-collected and patient-collected vaginal swab specimens. US clinical sites, including private and academic family practice, obstetric-gynecologic, family planning, public health, sexually transmitted infections (STI), and medical group clinics, and clinical research centers.

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

Clinical Performance Study:
A multicenter study was conducted using prospectively-collected patient- and clinician-collected vaginal swab samples from women ≥14 years who exhibited signs and/or symptoms of vaginitis. Twenty-one participating geographically diverse US clinical sites, including private and academic family practice, obstetric-gynecologic, family planning, public health, sexually transmitted infections (STI), and medical group clinics, and clinical research centers, obtained vaginal swab samples from 1519 symptomatic women.

For each symptomatic subject, two (2) vaginal swab samples (one patient-collected, one clinician-collected) were tested with the investigational Aptima CV/TV assay. Three vaginal swab samples were used for Candida and T. vaginalis culture and/or NAAT testing to determine Candida and T. vaginalis infection status. Candida species group and C. glabrata reference results were established based on a combination of Candida growth in culture, as well as PCR/bi-directional sequencing of both Aptima swab samples leftover after testing, for subjects with positive Candida culture results. For T. vaginalis, a patient infected status (PIS) was determined for each subject based on results from testing with an FDA-cleared molecular TV assay and an FDA-cleared culture- based TV test.

Contrived C. glabrata samples were also tested with the Aptima CV/TV assay to supplement clinical performance results in symptomatic subjects.

Of the 1519 subjects enrolled, 23 were not evaluable due to withdrawal (n = 17) or unknown or missing swabs or testing results (n = 6). The remaining 1496 subjects were evaluable for at least one analyte in at least one sample type.

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

Brief Description of Analytical (Non-Clinical) Studies

Analytical Sensitivity (LoD): Determined by testing a series of panels consisting of target organisms diluted in pooled negative clinical specimens or simulated vaginal swab matrix (SVSM). A minimum of 20 replicates of each panel member were tested with each of the two reagent lots, for a minimum of 40 replicates per panel member. Probit analysis was performed to generate the 95% predicted detection limit for each organism.
- Predicted Detection Limits: C. albicans: 4439 CFU/mL; C. glabrata: 41 CFU/mL; C. parapsilosis: 9416 CFU/mL; C. tropicalis: 811 CFU/mL; C. dubliniensis: 1176 CFU/mL; T. vaginalis: 0.0024 Cells/mL.
Analytical Inclusivity: Five strains of each Candida target organism were tested using lysate targeting 3X LoD for C. albicans, C. parapsilosis, C. tropicalis, C. dubliniensis, and C. glabrata in SVSM. Nine strains of T. vaginalis including a metronidazole resistant strain were tested with cell lysate targeting 3X LoD in SVSM.
- Results: The Aptima CV/TV assay was positive for all Candida strains tested at 3X LoD. Eight of the nine T. vaginalis strains, including the metronidazole resistant strain, were detected at 3X LoD. One strain of T. vaginalis was detected at 4X LoD.
Cross-Reactivity and Microbial Interference: Evaluated in the presence of closely related and non-targeted organisms. A panel consisting of 64 organisms and human cell lines was tested in SVSM in the absence or presence of 3X LoD C. albicans, C. glabrata or T. vaginalis.
- Results: No cross-reactivity or microbial interference was observed for any of the 64 organisms and human cell lines tested at the specified concentrations. (Note: Cross-reactivity with Candida famata was seen at concentrations higher than 5x10 CFU/mL).
Interference: Potentially interfering substances were tested in the Aptima CV/TV assay. Sensitivity performance was evaluated for C. albicans, C. glabrata, and T. vaginalis by spiking lysate at 3X LoD. Negative panels containing each substance were also evaluated for specificity.
- Results: No interference was observed with substances tested at specified concentrations (e.g., Whole Blood 5% V/V, Leukocytes 1x10^6 cells/mL, Mucus 5% V/V, Seminal Fluid 5% V/V). Certain substances (Tioconazole, Vaginal Moisturizing Gel, Glacial Acetic Acid) showed interference at higher concentrations but not at the tested concentrations.
Within Laboratory Precision: Evaluated on three Panther systems at one site. Three operators performed testing across 22 days and three reagent lots. Each operator performed two runs per day using a 7-member panel (three replicates each). Panel members were C. albicans, C. glabrata, or T. vaginalis in SVSM at Low and Moderate Positive concentrations, and a Negative panel.
- Results: Percent positivity for all low and moderate positive panels was 100% (95% CI: 97.7-100.0). Negative panel was 0% positive (95% CI: 0.0-2.3). Signal variability (TTime) showed good precision.
Co-Infection: Evaluated the ability to detect Candida species, C. glabrata, and T. vaginalis when more than one organism is present. Low concentration of one target lysate and high concentration of another target lysate in SVSM were tested in combination.
- Results: All testing resulted in 100% detection for both targets present, except for some combinations of low C. glabrata and high T. vaginalis, which upon further testing also showed 100% detection.

Brief Description of Clinical Studies

Clinical Performance Study: Multicenter study (21 US clinical sites) with 1496 evaluable symptomatic women (age 14-79, mean 35.3 ± 11.76 years). Tested clinician-collected and patient-collected vaginal swab samples. Reference results for Candida species group and C. glabrata based on culture and PCR/bi-directional sequencing. For T. vaginalis, Patient Infected Status (PIS) determined by FDA-cleared molecular TV assay and culture-based TV test. Supplemental testing with contrived C. glabrata samples.
- Key Results:
  - Candida species Group (Patient-collected): Sensitivity 92.9% (90.0-95.0), Specificity 91.0% (89.1-92.6), PPV 80.5% (77.4-83.4), NPV 97.0% (95.8-97.9).
  - Candida species Group (Clinician-collected): Sensitivity 91.7% (88.7-94.0), Specificity 94.9% (93.4-96.1), PPV 87.8% (84.8-90.4), NPV 96.6% (95.5-97.6).
  - C. glabrata (Patient-collected): Sensitivity 86.2% (75.1-92.8), Specificity 98.7% (98.0-99.2), PPV 73.5% (63.7-82.2), NPV 99.4% (99.0-99.7).
  - C. glabrata (Clinician-collected): Sensitivity 84.7% (73.5-91.8), Specificity 99.1% (98.4-99.5), PPV 79.4% (69.4-87.5), NPV 99.4% (98.9-99.7).
  - T. vaginalis (Patient-collected): Sensitivity 97.1% (92.9-98.9), Specificity 98.9% (98.2-99.4), PPV 90.7% (85.5-94.5), NPV 99.7% (99.2-99.9).
  - T. vaginalis (Clinician-collected): Sensitivity 96.5% (92.0-98.5), Specificity 95.1% (93.8-96.2), PPV 68.5% (63.2-73.8), NPV 99.6% (99.1-99.9).
  - Contrived C. glabrata samples: 100% agreement with known status for all samples, regardless of organism load.
Evaluation in Asymptomatic Women: Evaluated detection of Candida species group and C. glabrata in clinician-collected vaginal specimens from 171 asymptomatic women ≥ 14 years.
- Results: Positivity rates varied: Candida species group 21.1% (36/171), C. glabrata 8.8% (15/171). Detected in all ethnic categories except Asian.
Reproducibility: Evaluated at three US sites using seven panel members (one reagent lot, six operators, at least six days of testing).
- Results: Agreement with expected results was 100% for all panel members containing C. albicans, C. glabrata, or T. vaginalis (100% Agreement with 95% CI from 96.5-100 for positive samples and 99.3-100 for negative samples). Total %CV values for signal variability ranged from 5.64% to 6.77%.

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

Patient-collected, Candida species group:
- Sensitivity: 92.9% (90.0-95.0)
- Specificity: 91.0% (89.1-92.6)
- PPV: 80.5% (77.4-83.4)
- NPV: 97.0% (95.8-97.9)
Clinician-collected, Candida species group:
- Sensitivity: 91.7% (88.7-94.0)
- Specificity: 94.9% (93.4-96.1)
- PPV: 87.8% (84.8-90.4)
- NPV: 96.6% (95.5-97.6)
Patient-collected, C. glabrata:
- Sensitivity: 86.2% (75.1-92.8)
- Specificity: 98.7% (98.0-99.2)
- PPV: 73.5% (63.7-82.2)
- NPV: 99.4% (99.0-99.7)
Clinician-collected, C. glabrata:
- Sensitivity: 84.7% (73.5-91.8)
- Specificity: 99.1% (98.4-99.5)
- PPV: 79.4% (69.4-87.5)
- NPV: 99.4% (98.9-99.7)
Patient-collected, T. vaginalis:
- Sensitivity: 97.1% (92.9-98.9)
- Specificity: 98.9% (98.2-99.4)
- PPV: 90.7% (85.5-94.5)
- NPV: 99.7% (99.2-99.9)
Clinician-collected, T. vaginalis:
- Sensitivity: 96.5% (92.0-98.5)
- Specificity: 95.1% (93.8-96.2)
- PPV: 68.5% (63.2-73.8)
- NPV: 99.6% (99.1-99.9)
Aptima (C. glabrata Contrived Specimens):
- Low Positive (3x LOD): PPA 100 (88.6-100)
- Moderate Positive (10x LOD): PPA 100 (79.6-100)
- High Positive (20x LOD): PPA 100 (79.6-100)
- True Negative: NPA 100 (94.0-100)

Predicate Device(s)

BD MAX Vaginal Panel (DEN160001)

Reference Device(s)

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information

Not Found

Regulation Number and Section

N/A

Summary

0

Image /page/0/Picture/0 description: The image contains the logos of the Department of Health & Human Services and the Food and 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 includes the letters "FDA" in a blue square, followed by the words "U.S. FOOD & DRUG ADMINISTRATION" in blue text.

May 16, 2019

Hologic, Inc. Jeffrey Hergesheimer Regulatory Affairs Manager 10210 Genetic Center Drive San Diego, California 92121

Re: K190472

Trade/Device Name: Aptima CV/TV Assay Regulation Number: 21 CFR 866.3975 Regulation Name: Device that detects nucleic acid sequences from microorganisms associated with vaginitis and bacterial vaginosis Regulatory Class: Class II Product Code: PQA, NSU Dated: February 25, 2019 Received: February 26, 2019

Dear Jeffrey Hergesheimer:

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. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. 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

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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) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.htm); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; 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 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.

for

Uwe Scherf. Ph.D. Director Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K190472

Device Name Aptima CV/TV Assay

Indications for Use (Describe)

The Aptima CV/TV assay is an in vitro nucleic acid amplification of RNA from microorganisms associated with vulvovaginal candidiasis and trichomoniasis. The assay utilizes real time transcription-mediated amplification (TMA) to detect and qualitatively report results for the following organisms:

Candida species group (C. albicans, C. tropicalis, C. parapsilosis, C. dubliniensis)
Candida glabrata
Trichomonas vaginalis

The assay differentiates between Candida glabrata and the Candida species group (C spp) by targeting the RNA component of RNAse Pribonucleoprotein; the assay does not differentiate among C spp. For Trichomonas vaginalis, the assay targets ribosomal RNA (rRNA) and differentiates the results for Candida glabrata and C spp. The assay is intended to aid in the diagnosis of vulvovaginal candidiasis and trichomoniasis on the automated Panther system using clinician-collected and patient-collected vaginal swab specimens from females with a clinical presentation consistent with vaginitis or vulvovaginitis.

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

	☑ Prescription Use (Part 21 CFR 801 Subpart D)
	☐ Over-The-Counter Use (21 CFR 801 Subpart C)

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510(k) SUMMARY Aptima CV/TV Assay

I. SUBMITTER

Hologic, Inc. 10210 Genetic Center Drive San Diego, CA 92121

Contact Person: Jeffrey Hergesheimer, MS, RAC Regulatory Affairs Manager Jeffrey.Hergesheimer@Hologic.com Phone: (858) 410-8536 (858) 410-5557 Fax:
Date Prepared: February 20, 2019

II. DEVICE

Proprietary Name of Device:	Aptima CV/TV Assay
Classification Name:	Device that detects nucleic acid sequences from microorganisms associated with vaginitis and bacterial vaginosis
Regulation Number:	21 CFR 866.3975
Regulatory Class:	Class II
Product Code:	PQA, NSU

III. PREDICATE DEVICE

The predicate device is the BD MAX Vaginal Panel (DEN160001; approved 10/28/2016, BD Diagnostics).

IV. DEVICE DESCRIPTION

The Aptima CV/TV assay is an in vitro nucleic acid amplification test for the detection and quantitation of RNA from microorganisms associated with vulvovaginal candidiasis and trichomoniasis in women with a clinical presentation consistent with vaginitis or vulvovaginitis. The Aptima CV/TV assay utilizes the automated Panther system to provide qualitative results to aid in the diagnosis of vulvovaginal candidiasis and trichomoniasis.

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Principles of the Procedure

The Aptima CV/TV assay involves three main steps, all of which take place in a single tube on the Panther system: target capture, target amplification by TMA, and detection of the amplification products (amplicon) by fluorescent labeled probes (torches). The assay incorporates an internal control (IC) in every test to monitor nucleic acid capture, amplification and detection.

Specimens are collected in a tube containing specimen transport media (STM) that lyses the organisms, releases the RNA, and protects it from degradation during storage. When the assay is performed, capture oligonucleotides hybridize to highly conserved regions of the target RNA, if present, in the test specimen. The hybridized target is then captured onto magnetic microparticles that are separated from the specimen in a magnetic field. Wash steps remove extraneous components from the reaction tube.

Target amplification occurs via TMA, a transcription-based nucleic acid amplification method that utilizes two enzymes, Moloney murine leukemia virus (MMLV) reverse transcriptase and T7 RNA polymerase. The reverse transcriptase is used to generate a DNA copy of the target RNA sequence, adding a promoter sequence for T7 RNA polymerase. T7 RNA polymerase produces multiple copies of RNA amplicon from the DNA copy template.

Detection is achieved using single-stranded nucleic acid torches that are present during the amplification of the target and hybridize specifically to the amplicon in real time. Each torch has a fluorophore and a quencher. The quencher suppresses the fluorescence of the fluorophore when the torch is not hybridized to the amplicon. When the torch binds to the amplicon, the fluorophore is separated from the quencher and emits a signal at a specific wavelength when excited by a light source. The Panther system detects and discriminates between four fluorescent signals corresponding to C spp, C. glabrata, T. vaginalis and IC amplification products. The Panther system software uses an Aptima CV/TV assay-specific algorithm that interprets the amplification signal emergence times to generate a Positive or Negative status for each target organism in the sample.

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Assay Components

The Aptima CV/TV assay is provided as a 100-test kit. The Aptima CV/TV assay master kit contains 8 reagents, 1 calibrator, and 2 controls required for sample processing. There are 4 boxes that make up the assay master kit. Boxes 1 and 2 contain the Aptima CV/TV assay reagents packaged according to storage conditions. Box 3 contains the calibrator, and Box 4 contains the controls when provided as part of the master kit. The Aptima CV/TV Calibrator and Controls kit may also be procured separately if customers need additional calibrators or controls. A listing of the components that are required to perform the Aptima CV/TV assay are detailed in Table 1. In addition, there is one ancillary kit required to run the assay, and one collection kit utilized for collection of specimens (Table 2).

Box	Components Description
1	Amplification Reagent
1	Enzyme Reagent
1	Promoter Reagent
1	Internal Control
2	Amplification Reconstitution Solution
2	Enzyme Reconstitution Solution
2	Promoter Reconstitution Solution
2	Target Capture Reagent
3	Positive Calibrator
4	Negative Control
4	Positive Control

Table 1: Reagents Required to Perform the Aptima CV/TV Assay

							Table 2: Ancillarv and Collection Kits Required to Perform the Aptima CV/TV Assav
--	--	--	--	--	--	--	-----------------------------------------------------------------------------------	--	--

Aptima Assay Fluids Kit
Aptima Multitest Swab Specimen Collection Kit

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Instrumentation

The Aptima CV/TV assay has been designed for and validated on the Panther system. The Panther system is an integrated hardware and software system that together with the Aptima CV/TV assay fully automates all the steps necessary to perform the assay from sample preparation through amplification of nucleic acid, detection, data reduction and amplicon inactivation.

V. INDICATIONS FOR USE

The Aptima CV/TV assay is an in vitro nucleic acid amplification test for the detection of RNA from microorganisms associated with vulvovaginal candidiasis and trichomoniasis. The assay utilizes real time transcription-mediated amplification (TMA) to detect and qualitatively report results for the following organisms:

Candida species group (C. albicans, C. tropicalis, C. parapsilosis, C. dubliniensis)
Candida glabrata
Trichomonas vaginalis

The assay differentiates between Candida glabrata and the Candida species group (C spp) by targeting the RNA component of RNAse P ribonucleoprotein; the assay does not differentiate among C spp. For Trichomonas vaginalis, the assay targets ribosomal RNA (rRNA) and differentiates the result from results for Candida glabrata and C spp. The assay is intended to aid in the diagnosis of vulvovaginal candidiasis and trichomoniasis on the automated Panther system using clinician-collected and patient-collected vaginal swab specimens from females with a clinical presentation consistent with vaginitis or vulvovaginitis.

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VI. COMPARISON OF TECHNOLOGICAL CHARACTERISTICS WITH THE PREDICATE DEVICE

A comparison of the Aptima CV/TV assay to the predicate device, BD MAX Vaginal Panel, is summarized in Table 4 (similarities) and Table 5 (differences).

| Item | BD MAX Vaginal Panel
(DEN160001) | Aptima CV/TV Assay
(Subject Device) |
|--------------------|------------------------------------------------------------------------------------------------------|----------------------------------------|
| Patient Population | Women with a clinical presentation
of vaginitis, trichomoniasis, or
vulvovaginitis | Same |
| Specimen Types | Vaginal swabs in patients who are
symptomatic for vaginitis,
trichomoniasis, or vulvovaginitis | Same |
| Assay Controls | Incorporates an Internal Control in
every test. Uses external positive
and negative controls. | Same |

Table 4: Similarities Between Predicate Device and Subject Device

Table 5: Differences Between Predicate Device and Subject Device

| Item | BD MAX Vaginal Panel
(DEN160001) | Aptima CV/TV Assay
(Subject Device) |
|-----------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Organisms Detected | Lactobacillus (L. crispatus, and L. jensenii), Gardnerella vaginalis, Atopobium vaginae, Bacterial Vaginosis Associated Bacteria-2 (BVAB-2), Megasphaera-1, Candida (C. albicans, C. tropicalis, C. parapsilosis, C. dubliniensis), Candida glabrata, Candida krusei, Trichomonas vaginalis | Candida species (C. albicans, C. tropicalis, C. parapsilosis, C. dubliniensis); Candida glabrata; Trichomonas vaginalis |
| Platform/Technology
Principle of Operation | BD MAX System/
Real-time polymerase chain reaction (PCR) | Panther System/
Real-time Transcription Mediated Amplification (TMA) |
| Analyte | DNA | RNA |
| Assay Calibrators | None | Positive Calibrator |
| Intended Use | The BD MAX Vaginal Panel performed on the BD MAX | The Aptima CV/TV assay is an in vitro nucleic acid amplification test |
| Item | BD MAX Vaginal Panel
(DEN160001) | Aptima CV/TV Assay
(Subject Device) |
| | System is an automated qualitative in vitro diagnostic test for the direct detection of DNA targets from bacteria associated with bacterial vaginosis (qualitative results reported based on detection and quantitation of targeted organism markers), Candida species associated with vulvovaginal candidiasis, and Trichomonas vaginalis from vaginal swabs in patients who are symptomatic for vaginitis/vaginosis. The test utilizes real-time polymerase chain reaction (PCR) for the amplification of specific DNA targets and utilizes fluorogenic target-specific hybridization probes to detect and differentiate DNA from:
• Bacterial vaginosis markers (Individual markers not reported)
Lactobacillus spp. ( L. crispatus and L. jensenii )
Gardnerella vaginalis
Atopobium vaginae
Bacterial Vaginosis Associated Bacteria-2 (BVAB-2)
Megasphaera-1
• Candida spp. ( C. albicans, C. tropicalis, C. parapsilosis, C. dubliniensis )
• Candida glabrata
• Candida krusei
• Trichomonas vaginalis
The BD MAX Vaginal Panel is intended to aid in the diagnosis of vaginal infections in women with a clinical presentation consistent with bacterial vaginosis, vulvovaginal candidiasis and | for the detection of RNA from microorganisms associated with vulvovaginal candidiasis and trichomoniasis. The assay utilizes real time transcription-mediated amplification (TMA) to detect and qualitatively report results for the following organisms:
• Candida species group ( C. albicans, C. tropicalis, C. parapsilosis, C. dubliniensis )
• Candida glabrata
• Trichomonas vaginalis

The assay differentiates between Candida glabrata and the Candida species group (C spp) by targeting the RNA component of RNAse P ribonucleoprotein; the assay does not differentiate among C spp. For Trichomonas vaginalis , the assay targets ribosomal RNA (rRNA) and differentiates the result from results for Candida glabrata and C spp. The assay is intended to aid in the diagnosis of vulvovaginal candidiasis and trichomoniasis on the automated Panther system using clinician-collected and patient-collected vaginal swab specimens from females with a clinical presentation consistent with vaginitis or vulvovaginitis. |

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VII. PERFORMANCE DATA

The following performance data were provided in support of the substantial equivalence determination.

Brief Description of Analytical (Non-Clinical) Studies

The following analytical (non-clinical) studies were conducted to support the clearance of the Aptima CV/TV assay on the Panther system.

Analytical Sensitivity

The analytical sensitivity/LoD of the Aptima CV/TV assay was determined by testing a series of panels consisting of target organisms diluted in pooled negative clinical specimens or simulated vaginal swab matrix (SVSM). A minimum of 20 replicates of each panel member were tested with each of the two reagent lots, for a minimum of 40 replicates per panel member. Probit analysis was performed to generate the 95% predicted detection limit for each organism. The predicted detection limits are shown in Table 6.

Table 6: Limit of Detection of the Aptima CV/TV Assav
-------------------------------------------------------	--	--

Organism	Predicted Detection Limit	Concentration	Units
C. albicans	તે તે જેની જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામનાં પ્રાથમિક શાળા, પંચાયતઘર, આંગણવાડી તેમ જ દૂધની ડેરી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામનાં પ્રાથમિક શાળા, પંચાયતઘર, આંગણવાડી ત	4439	CFU/mL
C. glabrata	તે તે જેની જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામનાં પ્રાથમિક શાળા, પંચાયતઘર, આંગણવાડી તેમ જ દૂધની ડેરી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામનાં પ્રાથમિક શાળા, પંચાયતઘર, આંગણવાડી ત	41	CFU/mL
C. parapsilosis1	તે તે જે જે જે જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામમાં પ્રાથમિક શાળા, પંચાયતઘર, આંગણવાડી તેમ જ દૂધની ડેરી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામનાં પ્રાથમિક શાળા, પંચાયતઘર, આંગણવા	9416	CFU/mL
C. tropicalis4	તે જેન્જ	811	CFU/mL
C. dubliniensis4	તે તે જેની જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામનાં પ્રાથમિક શાળા, પંચાયતઘર, આંગણવાડી તેમ જ દૂધની ડેરી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામનાં પ્રાથમિક શાળા, પંચાયતઘર, આંગણવાડી ત	1176	CFU/mL
T. vaginalis	તે જેને જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામનાં લોકોનો મુખ્ય વ્યવસાય ખેતી, ખેતમજૂરી તેમ જ પશુપાલન છે. આ ગામમાં પ્રાથમિક શાળા, પંચાયતઘર, આંગણવાડી તેમ જ દૂધની ડેરી જેવી સવલતો પ્	0.0024	Cells/mL

¹Tested in simulated vaginal swab matrix

Analytical Inclusivity

Five strains of each Candida target organism were tested using lysate targeting 3X LoD for C. albicans, C. parapsilosis, C. tropicalis, C. dubliniensis, and C. glabrata in SVSM. Nine strains of T. vaginalis including a metronidazole resistant strain were tested with cell lysate targeting 3X LoD in SVSM. The Aptima CV/TV assay was positive for all Candida strains

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tested at 3X LoD. Eight of the nine T. vaginalis strains, including the metronidazole resistant strain, were detected at 3X LoD. One strain of T. vaginalis was detected at 4X LoD.

Cross-Reactivity and Microbial Interference

Cross-reactivity and microbial interference with the Aptima CV/TV assay was evaluated in the presence of closely related and non-targeted organisms. A panel consisting of 64 organisms and human cell lines (Table 7) was tested in SVSM in the absence or presence of 3X LoD C. albicans, C. glabrata or T. vaginalis. No cross-reactivity or microbial interference was observed for any of the 64 organisms and human cell lines tested in the Aptima CV/TV assay at the following concentrations.

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Microorganism	Concentration	Microorganism	Concentration
Acinetobacter lwoffii	1x106 CFU/mL	Herpes simplex virus I	1x104 TCID 50/mL
Actinomyces israelii	1x106 CFU/mL	Herpes simplex virus II	1x104 TCID 50/mL
Alcaligenes faecalis	1x106 CFU/mL	Klebsiella pneumoniae	1x106 CFU/mL
Atopobium vaginae	1x106 CFU/mL	Lactobacillus acidophilus	1x106 CFU/mL
Bacteroides fragilis	1x106 CFU/mL	Lactobacillus crispatus	1x106 CFU/mL
Bifidobacterium adolescentis	1x106 CFU/mL	Lactobacillus gasseri	1x106 CFU/mL
BVAB-11	1x106 copies/mL	Lactobacillus iners	1x106 CFU/mL
BVAB-21	1x106 copies/mL	Lactobacillus jensenii	1x106 CFU/mL
Campylobacter jejuni	1x106 CFU/mL	Lactobacillus mucosae	1x106 CFU/mL
Candida catenulata	1x106 CFU/mL	Leptotrichia buccalis	1x106 CFU/mL
Candida famata2	5x105 CFU/mL	Listeria monocytogenes	1x106 CFU/mL
Candida guilliermondii	1x106 CFU/mL	Megasphaera Type 11	1x106 copies/mL
Candida haemulonii	1x106 CFU/mL	Mobiluncus curtisii	1x106 CFU/mL
Candida inconspicua	1x106 CFU/mL	Mycoplasma genitalium	1x106 CFU/mL
Candida kefyr	1x106 CFU/mL	Mycoplasma hominis	1x106 CFU/mL
Candida krusei	1x106 CFU/mL	Neisseria gonorrhoeae	1x106 CFU/mL
Candida lusitaniae	1x106 CFU/mL	Peptostreptococcus magnus	1x106 CFU/mL
Candida norvegica	1x106 CFU/mL	Pentatrichomonas hominis	1x105 cells/mL
Candida orthopsilosis	1x106 CFU/mL	Pichia fermentans	1x106 CFU/mL
Chlamydia trachomatis	1x106 IFU/mL	Prevotella bivia	1x106 CFU/mL
Clostridium difficile	1x106 CFU/mL	Propionibacterium acnes	1x106 CFU/mL
Corynebacterium genitalium	1x106 CFU/mL	Proteus vulgaris	1x106 CFU/mL
Cryptococcus neoformans	1x106 CFU/mL	SiHa cells	1x104 cells/mL
Eggerthella lenta	1x106 CFU/mL	Sneathia amnii	1x106 CFU/mL
Enterobacter cloacae	1x106 CFU/mL	Staphylococcus aureus	1x106 CFU/mL
Enterococcus faecalis	1x106 CFU/mL	Staphylococcus epidermidis	1x106 CFU/mL
Escherichia coli	1x106 CFU/mL	Streptococcus agalactiae	1x106 CFU/mL
Fusobacterium nucleatum	1x106 CFU/mL	Streptococcus pyogenes	1x106 CFU/mL
Gardnerella vaginalis	1x106 CFU/mL	Treponema pallidum2	1x106 copies/mL
Haemophilus ducreyi	1x106 CFU/mL	Trichomonas tenax	1x105 cells/mL
HeLa cells	1x104 Cells/mL	Ureaplasma parvum	1x106 CFU/mL
HIV	1x105 copies/mL	Ureaplasma urealyticum	1x106 CFU/mL

Table 7: Cross-Reactivity and Microbial Interference Panel

CFU = Colony Forming Units; IFU = Inclusion Forming Units; TCID50 = Median Tissue Culture Infectious Dose 1 In Vitro Transcript tested.

2 Cross-reactivity with Candida famata was seen at concentrations higher than 5x10 °CFU/mL.

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Interference

Potentially interfering substances were tested in the Aptima CV/TV assay. Panels were built in SVSM and evaluated for potential effects on assay sensitivity and specificity. Sensitivity performance was evaluated separately for C. albicans, C. glabrata, and T. vaginalis by spiking lysate at 3X LoD. Negative panels containing each substance were also evaluated for specificity.

No interference was observed in the presence of the following exogenous and endogenous substances tested at the concentrations listed in Table 8.

Substance	Final Concentration1
Whole Blood	5% V/V
Leukocytes	1x106 cells/mL
Mucus	5% V/V
Seminal Fluid	5% V/V
Contraceptive Foam	5% W/V
Contraceptive Film	5% W/V
Tioconazole2	2% W/V
Douche	5% W/V
Progesterone	5% W/V
Estradiol	5% W/V
Acyclovir	5% W/V
Metronidazole	5% W/V
Hemorrhoidal Cream	5% W/V
Vaginal Moisturizing Gel3	0.5% W/V
Lubricant	5% V/V
Spermicide	5% W/V
Anti-fungal	5% W/V
Deodorant/Spray	5% W/V
Glacial Acetic Acid4	4% V/V
Vagisil Cream	5% W/V

Table 8: Interfering Substances Panel

W/V = weight by volume; V/V = volume by volume

1 Final Concentration represents final concentration in the sample when tested on the Panther instrument.

2 Tioconazole 6.5% Ointment: Interference was observed at ≥ 3% W/V for all analytes. No interference was observed at 2% W/V for all analytes.

3 Vaginal Moisturizing Gel: Interference was observed at ≥ 1% W/V for C. albicans, 5% W/V for C. glabrata, and ≥ 3% W/V for T. vaginalis. No interference was observed at 0.5% W/V for C. albicans, 4% W/V for C. glabrata, and 2% W/V for T. vaginalis.

4 Glacial Acetic Acid: Interference was observed at 5% V/V for C. albicans. No

interference was observed at 4% V/V for C. albicans, 5% V/V for C. glabrata, and 5% V/V for T. vaginalis.

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Within Laboratory Precision

Within Lab Precision was evaluated on three Panther systems at one site. Three operators performed testing across 22 days and three reagent lots. Each operator performed two runs per day using a 7-member panel. Each run consisted of three replicates of each panel member. The panel members were made with C. albicans, C. glabrata or T. vaginalis in SVSM. The six positive panel members targeted C. albicans at Low and Moderate Positive, C. glabrata at Low and Moderate Positive, and T. vaginalis at Low and Moderate Positive. One Negative panel member contained matrix with no added target analytes.

The CV/TV percent positive results are presented in Table 9. Signal variability (TTime) of the Aptima CV/TV assay was also calculated for analyte positive panel members. Variability calculated between instruments, between operators, between days, between days, between runs, within runs, and overall, is shown in Table 10.

| Panel (analyte composition) | Positive / Total n | Expected Positivity | Percent Positivity
(95% CI) |
|----------------------------------|--------------------|---------------------|--------------------------------|
| Negative (SVSM) | 0/162 | 0% | 0 (0.0-2.3) |
| Low Positive (C. albicans) | 162/162 | ≥95% | 100 (97.7-100.0) |
| Low Positive (C. glabrata) | 162/162 | ≥95% | 100 (97.7-100.0) |
| Low Positive (T. vaginalis) | 162/162 | ≥95% | 100 (97.7-100.0) |
| Moderate Positive (C. albicans) | 162/162 | ≥95% | 100 (97.7-100.0) |
| Moderate Positive (C. glabrata) | 162/162 | ≥95% | 100 (97.7-100.0) |
| Moderate Positive (T. vaginalis) | 162/162 | ≥95% | 100 (97.7-100.0) |

Table 9: Precision - Agreement of Aptima CV/TV Assav with Expected Results

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| | | Between
Days | | Between
Instruments | | Between
Operators | | Between
Lots | | Between
Runs | | Within
Run | | Total | | |
|-----------------------------------|-----|-----------------|------|------------------------|------|----------------------|------|-----------------|------|-----------------|------|---------------|------|--------|------|--------|
| Panel
Description | N | Mean
TTime | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) |
| C. albicans
Low Positive | 162 | 14.96 | 0.12 | 0.82 | 0.00 | 0.00 | 0.24 | 1.59 | 0.54 | 3.58 | 0.23 | 1.52 | 0.28 | 1.84 | 0.70 | 4.66 |
| C. glabrata
Low Positive | 162 | 21.07 | 0.00 | 0.00 | 0.15 | 0.69 | 0.25 | 1.18 | 0.14 | 0.65 | 0.19 | 0.89 | 0.40 | 1.91 | 0.55 | 2.59 |
| T. vaginalis
Low Positive | 162 | 24.09 | 0.00 | 0.00 | 0.33 | 1.38 | 0.22 | 0.93 | 0.01 | 0.05 | 0.21 | 0.87 | 0.59 | 2.46 | 0.75 | 3.09 |
| C. albicans
Moderate Positive | 162 | 14.62 | 0.11 | 0.72 | 0.00 | 0.00 | 0.22 | 1.47 | 0.43 | 2.95 | 0.26 | 1.77 | 0.24 | 1.62 | 0.60 | 4.14 |
| C. glabrata
Moderate Positive | 162 | 20.63 | 0.00 | 0.00 | 0.00 | 0.00 | 0.26 | 1.27 | 0.31 | 1.50 | 0.26 | 1.25 | 0.52 | 2.51 | 0.71 | 3.42 |
| T. vaginalis
Moderate Positive | 162 | 22.73 | 0.00 | 0.00 | 0.12 | 0.54 | 0.24 | 1.08 | 0.18 | 0.80 | 0.28 | 1.23 | 0.41 | 1.79 | 0.59 | 2.61 |

Table 10: Signal Variability of the Aptima CV/TV Assay by Analyte Positive Panel Member

CV = Coefficient of variation

Note: Variability from some factors may be numerically negative. This can occur if the variability due to those factors is very small. In these cases, SD and CV are shown as 0.00.

Co-Infection

A co-infection study evaluated the ability of the Aptima CV/TV assay to detect Candida species, C. glabrata, and T. vaginalis when more than one organism is present in the same specimen. Low concentration of one target lysate and high concentration of another target lysate in SVSM were tested in combination. Panel composition and concentrations are listed in Table 11. All testing resulted in 100% detection for both targets present except for the combinations of low C. glabrata (3X LoD) and high T. vaginalis (1x10 cells/mL or 1x10 cells/mL). Further testing was conducted and resulted in 100% detection for the combination of low C. glabrata (3X LoD) and high T. vaginalis (1x103 cells/mL).

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Panel Member	C. albicans Concentration	C. glabrata Concentration	T. vaginalis Concentration
C. albicans Low; C. glabrata High	13317 CFU/mL1	1x106 CFU/mL	N/A
C. albicans Low; T. vaginalis High	13317 CFU/mL1	N/A	1x105 cells/mL
C. glabrata Low; T. vaginalis High	N/A	123 CFU/mL2	1x103 cells/mL
C. albicans High; C. glabrata Low	1x106 CFU/mL	123 CFU/mL2	N/A
C. albicans High; T. vaginalis Low	1x106 CFU/mL	N/A	0.0072 cells/mL3
C. glabrata High; T. vaginalis Low	N/A	1x106 CFU/mL	0.0072 cells/mL3

Table 11: Co-Infection Panel

CFU = Colony Forming Units 13X LoD C. albicans. 2 3X LoD C. glabrata. 3 3X LoD T. vaginalis.

Brief Description of Clinical Studies

Clinical testing of the Aptima CV/TV assay on the Panther system included performance and reproducibility testing. Substantial equivalence is based in part on the performance study.

Clinical Performance Study

A study was performed to demonstrate clinical performance characteristics for the Aptima CV/TV assay. A multicenter study was conducted using prospectively-collected patient- and clinician-collected vaginal swab samples from women ≥14 years who exhibited signs and/or symptoms of vaginitis. Twenty-one participating geographically diverse US clinical sites, including private and academic family practice, obstetric-gynecologic, family planning, public health, sexually transmitted infections (STI), and medical group clinics, and clinical research centers, obtained vaginal swab samples from 1519 symptomatic women.

For each symptomatic subject, two (2) vaginal swab samples (one patient-collected, one clinician-collected) were tested with the investigational Aptima CV/TV assay. Three vaginal swab samples were used for Candida and T. vaginalis culture and/or NAAT testing to determine Candida and T. vaginalis infection status. Candida species group and C. glabrata reference results were established based on a combination of Candida growth in culture, as well as PCR/bi-directional sequencing of both Aptima swab samples leftover after testing, for subjects with positive Candida culture results. For T. vaginalis, a patient infected status (PIS) was

16

determined for each subject based on results from testing with an FDA-cleared molecular TV assay and an FDA-cleared culture- based TV test.

Contrived C. glabrata samples were also tested with the Aptima CV/TV assay to supplement clinical performance results in symptomatic subjects.

The clinical performance of the Aptima CV/TV assay was estimated relative to the Candida species group and C. glabrata reference results and T. vaginalis PIS; sensitivity, specificity, PPV, and NPV were calculated for each analyte and Aptima sample type, along with corresponding 2-sided 95% Cls. Agreement with known positive or known negative status was calculated for contrived C. glabrata samples.

Of the 1519 subjects enrolled, 23 were not evaluable due to withdrawal (n = 17) or unknown or missing swabs or testing results (n = 6). The remaining 1496 subjects were evaluable for at least one analyte in at least one sample type. Table 12 shows the demographic and baseline clinical characteristics of evaluable subjects.

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	Symptomatic
Total, N	1496
Age, years
Mean ± SD	$35.3 \pm 11.76$
Median	33.0
Range	14-79
Age category (years), n (%)
14-17	5 (0.3)
18-29	554 (37.0)
30-39	480 (32.1)
40-49	247 (16.5)
>50	210 (14.0)
Race/Ethnicity, n (%)
Asian	73 (4.9)
Black or African American	752 (50.3)
White (Hispanic or Latino)	268 (17.9)
White (Not Hispanic or Latino)	339 (22.7)
Other¹	64 (4.3)

Table 12: Demographics of Evaluable Subjects in the Aptima CV/TV Assay Evaluation

1Includes patient-reported other, mixed, and unknown races

From the 1496 evaluable symptomatic subjects, results from 1477 patient-collected Aptima vaginal swab samples (98.7%, 1477/1496) and 1485 clinician-collected vaginal swab samples (99.3%, 1485/1496) were included in the analyses for Candida species group. 1475 patientcollected Aptima vaginal swab samples (98.6%, 1475/1496) and 1483 clinician-collected vaginal swab samples (99.1%, 1483/1496) were included in the analyses for C. glabrata, and 1433 patient-collected Aptima vaginal swab samples (95.8%, 1433/1496) and 1438 clinician-collected vaginal swab samples (96.1%, 1438/1496) were included in the analyses for T. vaginalis. Performance characteristics for detection of Candida species group. C. glabrata, and T. vaginalis infection for patient-collected and clinician-collected vaginal swab samples were calculated overall (see Tables 13 through 15) and by race/ethnicity (see Tables 16 through 18).

Anticipated low prevalence of C. glabrata triggered the supplemental assessment of the assay using contrived C. glabrata samples. For contrived C. glabrata samples, the agreement with known status was 100% for all samples (see Table 19), regardless of the organism load. This confirmed that the performance of the investigational assay for the detection of C. glabrata was acceptable.

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Table 13: Aptima CV/TV Assay Performance for Candida species Group Relative to Candida Infection Status in Symptomatic Subjects

| Specimen
Type | N | TP | FP | TN | FN | Prevalence1 | Sensitivity
(95% CI)2 | Specificity
(95% CI)2 | PPV
(95% CI)3 | NPV
(95% CI)3 |
|-------------------------|------|-----|----|------|----|-------------|--------------------------|--------------------------|------------------|------------------|
| Patient-
collected | 1477 | 392 | 95 | 960 | 30 | 28.6 | 92.9 (90.0-95.0) | 91.0 (89.1-92.6) | 80.5 (77.4-83.4) | 97.0 (95.8-97.9) |
| Clinician-
collected | 1485 | 389 | 54 | 1007 | 35 | 28.6 | 91.7 (88.7-94.0) | 94.9 (93.4-96.1) | 87.8 (84.8-90.4) | 96.6 (95.5-97.6) |

FN = false negative, FP = false positive, NPV = negative predictive value, TP = true positive, TP = true positive, TN = true negative, 'Study prevalence reported. 'Score C. "PPV 95% Cl computed from the exact 95% CI for positive likelihood ratio; NPV 95% Cl computed from the 95% for the negative likelihood ratio

| Specimen
Type | N | TP | FP | TN | FN | Prevalence1 | Sensitivity
(95% CI)2 | Specificity
(95% CI)2 | PPV
(95% CI)3 | NPV
(95% CI)3 |
|-------------------------|------|----|-----|------|----|-------------|--------------------------|--------------------------|------------------|------------------|
| Patient-
collected | 1475 | 50 | 184 | 1399 | 85 | 3.9 | 86.2 (75.1-92.8) | 98.7 (98.0-99.2) | 73.5 (63.7-82.2) | 99.4 (99.0-99.7) |
| Clinician-
collected | 1483 | 50 | 136 | 1411 | 97 | 4.0 | 84.7 (73.5-91.8) | 99.1 (98.4-99.5) | 79.4 (69.4-87.5) | 99.4 (98.9-99.7) |

Table 14 . Antima (W/TV Assay Performance for Candida Infection Status in Sumptomatic Subjects

FN = false negative, FP = false positive value, PPV = positive predictive value, TP = true positive, TN = true negative, TN = true negative

'Study prevalence reported. 'Score C. "PPV 95% CI computed fikelihood rato; NPV 95% Cl computed from the 95% for the negative likelihood ratio

4 Of the 18 samples with false positive results, 2 showed low (32+) growth, and 14 showed no growth of C. glabrato on chromogenic agar.

5 Of the 8 samples with false negative results, 7 showed high (4+) growth of C. glabruta on chromogenic agar.

® Of the 13 samples with false positive results, 2 showed low (22+) growth, and 9 showed no growth of C. glabrato on chromogenc agar.

7 All 9 samples with false negative results showed no growth of C. glabrata on chromogenic agar.

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| Specimen
Type | N | TP | FP | TN | FN | Prevalence1 | Sensitivity
(95% CI)2 | Specificity
(95% CI)2 | PPV
(95% CI)3 | NPV
(95% CI)3 |
|-------------------------|------|-----|-----|------|----|-------------|--------------------------|--------------------------|------------------|------------------|
| Patient-
collected | 1433 | 136 | 144 | 1279 | 45 | 9.8 | 97.1 (92.9-98.9) | 98.9 (98.2-99.4) | 90.7 (85.5-94.5) | 99.7 (99.2-99.9) |
| Clinician-
collected | 1438 | 137 | 636 | 1233 | 57 | 9.9 | 96.5 (92.0-98.5) | 95.1 (93.8-96.2) | 68.5 (63.2-73.8) | 99.6 (99.1-99.9) |

Table 15: Aptima CV/TV Assay Performance for 7. vagualis Relative to the Patient Infected Status in Symptomatic Subiects

FN = false negative, FP = false positive, NPV = negative predictive value, TP = true positive, TP = true positive, TN = true negative,

'Study prevalence reported. 'Score CI. "PPV 95% CI computed ikelihood rato; NPV 95% CI computed from the 95% for the negative likelihood ratio

4 Of the 14 samples with false positive results, 8 were positive with a second FDA-cleared TV NAAT.

5 Of the 4 samples with false negative results, 3 were negative with a second FDA-cleared TV NAAT.

6 Of the 63 samples with false positive results, 56 were positive with a second FDA-cleared TV NAAT.

7 Of the 5 samples with false negative results, 3 were negative with a second FDA-cleared TV NAAT.

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| Race/Ethnicity | N | TP | FP | TN | FN | Prev¹
(%) | Sensitivity
(95% CI)² | Specificity
(95% CI)² | PPV
(95% CI)³ | NPV
(95% CI)³ |
|---------------------|-----------------------------|-----|-----|----|-----|--------------|--------------------------|--------------------------|------------------|------------------|
| Patient-collected | | | | | | | | | | |
| | Asian | 71 | 18 | 5 | 48 | 0 | 25.4 | 100 (82.4-100) | 90.6 (79.7-95.9) | 78.3 (62.2-91.6) |
| | Black / African American | 745 | 207 | 55 | 462 | 21 | 30.6 | 90.8 (86.3-93.9) | 89.4 (86.4-91.7) | 79.0 (74.7-83.1) |
| | White (Hispanic/Latino) | 265 | 71 | 19 | 170 | 5 | 28.7 | 93.4 (85.5-97.2) | 89.9 (84.8-93.5) | 78.9 (71.3-85.9) |
| | White (Not Hispanic/Latino) | 332 | 75 | 12 | 242 | 3 | 23.5 | 96.2 (89.3-98.7) | 95.3 (91.9-97.3) | 86.2 (78.8-92.1) |
| | Other⁴ | 64 | 21 | 4 | 38 | 1 | 34.4 | 95.5 (78.2-99.2) | 90.5 (77.9-96.2) | 84.0 (69.4-94.6) |
| Clinician-collected | | | | | | | | | | |
| | Asian | 73 | 19 | 3 | 51 | 0 | 26.0 | 100 (83.2-100) | 94.4 (84.9-98.1) | 86.4 (69.6-96.8) |
| | Black / African American | 747 | 206 | 31 | 489 | 21 | 30.4 | 90.7 (86.3-93.9) | 94.0 (91.7-95.8) | 86.9 (82.7-90.5) |
| | White (Hispanic/Latino) | 265 | 71 | 12 | 177 | 5 | 28.7 | 93.4 (85.5-97.2) | 93.7 (89.2-96.3) | 85.5 (77.9-91.6) |
| | White (Not Hispanic/Latino) | 336 | 73 | 6 | 250 | 7 | 23.8 | 91.3 (83.0-95.7) | 97.7 (95.0-98.9) | 92.4 (85.4-96.9) |
| | Other⁴ | 64 | 20 | 2 | 40 | 2 | 34.4 | 90.9 (72.2-97.5) | 95.2 (84.2-98.7) | 90.9 (75.6-98.6) |

Table 16: Aptima CVTV Assay Performance for Candida species Group Relative to Candida Infection Status in Symptomatic Subjects, by Race/Ethnicity

FN = false negative, FP = false positive, NPV = nesitive value, PPV = positive value, Prev = prevalence TP = true positive, TV = true negative

Study prevalence reported. 35cre Cl. - PPV 95% CI for positive likelihood ratio; NPV 95% CL computed from the 95% for the negative likelihood ratio. 4Includes patient-reported other, mixed, and unknown races.

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Race/Ethnicity	N	TP	FP	TN	FN	Prev¹ (%)	Sensitivity (95% CI)²	Specificity (95% CI)²	PPV (95% CI)³	NPV (95% CI)³
Patient-collected
Asian	71	3	1	67	0	4.2	100 (43.9-100)	98.5 (92.1-99.7)	75.0 (27.6-99.2)	100 (97.0-100)
Black / African American	744	24	9	704	7	4.2	77.4 (60.2-88.6)	98.7 (97.6-99.3)	72.7 (58.0-85.0)	99.0 (98.2-99.6)
White (Hispanic/Latino)	264	7	2	254	1	3.0	87.5 (52.9-97.8)	99.2 (97.2-99.8)	77.8 (49.1-97.0)	99.6 (98.4- 100)
White (Not Hispanic/Latino)	332	13	5	314	0	3.9	100 (77.2-100)	98.4 (96.4-99.3)	72.2 (53.0-88.9)	100 (99.0-100)
Other⁴	64	3	1	60	0	4.7	100 (43.9-100)	98.4 (91.3-99.7)	75.0 (27.9-99.2)	100 (96.6- 100)
Clinician-collected
Asian	72	3	0	69	0	4.2	100 (43.9-100)	100 (94.7-100)	100 (44.8-100)	100 (97.0-100)
Black / African American	747	23	9	707	8	4.1	74.2 (56.8-86.3)	98.7 (97.6-99.3)	71.9 (57.1-84.7)	98.9 (98.1-99.5)
White (Hispanic/Latino)	264	7	1	255	1	3.0	87.5 (52.9-97.8)	99.6 (97.8-99.9)	87.5 (56.5-99.5)	99.6 (98.4- 100)
White (Not Hispanic/Latino)	336	14	3	319	0	4.2	100 (78.5-100)	99.1 (97.3-99.7)	82.4 (61.7-95.8)	100 (99.0-100)
Other⁴	64	3	0	61	0	4.7	100 (43.9-100)	100 (94.1-100)	100 (44.9-100)	100 (96.6-100)

Table 17: Aptima CV/TV Assay Performance for C. glabrata Relative to Candida Infection Status in Symptomatic Subjects, by Race/Ethnicity

FN = false negative, FP = false positive, NPV = nesitive value, PPV = positive value, Prev = prevalence TP = true positive, TV = true negative

Study prevalence reported. 3 Score CI. PPV 95% CI for positive likelihood ratio; NPV 95% CI computed from the 95% for the negative likelihood ratio. 4Includes patient-reported other, mixed, and unknown races.

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Race/Ethnicity	N	TP	FP	TN	FN	Prev¹ (%)	Sensitivity (95% CI)²	Specificity (95% CI)²	PPV (95% CI)³	NPV (95% CI)³
Patient-collected
Asian	66	4	0	62	0	6.1	100 (51.0-100)	100 (94.2-100)	100 (52.8-100)	100 (96.3-100)
Black / African American	724	99	8	615	2	14.0	98.0 (93.1-99.5)	98.7 (97.5-99.3)	92.5 (86.5-96.5)	99.7 (98.9-100)
White (Hispanic/Latino)	258	16	5	236	1	6.6	94.1 (73.0-99.0)	97.9 (95.2-99.1)	76.2 (58.6-90.1)	99.6 (98.0-100)
White (Not Hispanic/Latino)	324	12	1	310	1	4.0	92.3 (66.7-98.6)	99.7 (98.2-99.9)	92.3 (69.6-99.7)	99.7 (98.5-100)
Other⁴	61	5	0	56	0	8.2	100 (56.6-100)	100 (93.6-100)	100 (58.3-100)	100 (95.5-100)
Clinician-collected
Asian	67	4	1	62	0	6.0	100 (51.0-100)	98.4 (91.5-99.7)	80.0 (40.3-99.4)	100 (96.3-100)
Black / African American	727	101	42	582	2	14.2	98.1 (93.2-99.5)	93.3 (91.0-95.0)	70.6 (64.3-76.6)	99.7 (98.8-100)
White (Hispanic/Latino)	257	16	12	228	1	6.6	94.1 (73.0-99.0)	95.0 (91.5-97.1)	57.1 (43.0-71.3)	99.6 (98.0-100)
White (Not Hispanic/Latino)	326	11	8	305	2	4.0	84.6 (57.8-95.7)	97.4 (95.0-98.7)	57.9 (39.9-75.8)	99.3 (98.1-99.9)
Other⁴	61	5	0	56	0	8.2	100 (56.6-100)	100 (93.6-100)	100 (58.3-100)	100 (95.5-100)

Table 18: Aptima CV/TV Assay Performance for 7. vaginalis Relative to Patient Infected Status in Symptomatic Subjects, by Race/Ethnicity

FN = false negative, FP = false positive, NPV = nositive predictive value, Prev = positive value, Prev = prevalence TP = true negative

Study prevalence reported. - Score CI. - PPV 95% CI for positive likelihood ratio; NPV 95% CI computed from the 95% for the negative likelihood ratio. 4Includes patient-reported other, mixed, and unknown races.

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| | N | Aptima
C. glabrata
Positive | Aptima
C. glabrata
Negative | PPA
(95% CI)1 | NPA
(95% CI)1 |
|-----------------------------|----|-----------------------------------|-----------------------------------|------------------|------------------|
| Low Positive (3× LOD) | 30 | 30 | 0 | 100 (88.6-100) | NC |
| Moderate Positive (10× LOD) | 15 | 15 | 0 | 100 (79.6-100) | NC |
| High Positive (20× LOD) | 15 | 15 | 0 | 100 (79.6-100) | NC |
| True Negative | 60 | 0 | 60 | NC | 100 (94.0-100) |

Table 19: Performance of the Aptima CV/TV Assay with C. glabrata Contrived Specimens

NC = Not Calculated, NPA = negative percent agreement, PPA = positive percent agreement ¹Score CI

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Evaluation of the Aptima CV/TV Assay in Asymptomatic Women

Although the Aptima CV/TV assay is not intended for testing specimens from asymptomatic women, presence of Candida species and C. glabrata as colonizing normal flora has been reported in this population. The Aptima CV/TV assay was evaluated for detection of Candida species group (C spp) and C. glabrata with clinician-collected vaginal specimens collected from 171 asymptomatic women ≥ 14 years. Aptima CV/TV assay C spp and C. glabrata targets were detected with rates varying from 8.8% for C. glabrata to 21.1% for C spp. Results from the evaluation are presented in Table 20 below which also includes results for the most prevalent ethnic groups enrolled. C spp and C. glabrata were detected in all ethnic categories, except for Asian.

	% Positivity (# positive/# tested with valid results)
	Candida species group	Candida glabrata
All	21.1% (36/171)	8.8% (15/171)
Asian	0.0% (0/5)	0.0% (0/5)
Black / African
American	28.0% (21/75)	12.0% (9/75)
White (Hispanic/Latino)	17.1% (7/41)	4.9% (2/41)
White
(Not Hispanic/Latino)	11.6% (5/43)	7.0% (3/43)
Other1	42.9% (3/7)	14.3% (1/7)

Table 20: Positivity of CV/TV Infection as Determined by the Aptima CV/TV Assav in Asymptomatic Subjects

Reproducibility

Aptima CV/TV assay reproducibility was evaluated at three US sites using seven panel members. Testing was performed using one lot of assay reagents and six operators (two at each site). At each site, testing was performed for at least six days. Each run had three replicates of each panel member.

The panel members were made using a simulated vaginal swab matrix ('SVSM', which contains specimen transport media (STM) spiked with simulated vaginal fluid) negative for Candida species (including C. glabrata) and T. vaginalis. Six panel members contained cell lysates of

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one of the following organisms: C. albicans (used as a representative of the Candida species group), C. glabrata, or T. vaginalis. Low positive and moderate positive concentrations of each analyte were tested. One negative panel member contained only the matrix with no added target analytes.

The agreement with expected results was 100% for all panel members containing C. albicans, C. glabrata, or T. vaginalis, as shown in Table 21.

Target	Concentration	Agreed/n	Agreement, % (95% CI)
C. albicans	Negative1	536/536	100 (99.3-100)
C. albicans	Low Positive2	108/108	100 (96.6-100)
C. albicans	Mod Positive3	107/107	100 (96.5-100)
C. glabrata	Negative1	538/538	100 (99.3-100)
C. glabrata	Low Positive2	106/106	100 (96.5-100)
C. glabrata	Mod Positive3	107/107	100 (96.5-100)
T. vaginalis	Negative1	536/536	100 (99.3-100)
T. vaginalis	Low Positive2	108/108	100 (96.6-100)
T. vaginalis	Mod Positive3	107/107	100 (96.5-100)

Table 21: Agreement of Aptima CV/TV Assay Results with Expected Results

CI = Score confidence interval, Mod=moderate, Neg = negative, Pos = positive

1 Includes all panel members without the target present.

2 Target concentration for low positive parel methods > Cg5 for C. albicans and 2× LOD for C. glabrata and T. vaginalis.

3 Target concentration for moderate positive panel members is:3× C, albicans and3× LOD for C. glabrata and T. vaginalis.

Across organisms/panel members, the total %CV values ranged from 5.64% to 6.77%; total SD values were ≤1.59. For most panel members, the "between sites", "between runs", and "within runs" factors were the largest contributors to total variability; for all other sources of variation, SD values were ≤0.30 (%CV values were ≤1.46%), as shown in Table 22.

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| | | Between
Sites | | Between
Operators | | Between
Days | | Between
Runs | | Within
Runs | | Total | | |
|-------------------|-----|------------------|------|----------------------|------|-----------------|------|-----------------|------|----------------|------|--------|------|--------|
| Panel Description | N | Mean
TTime1 | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) |
| Calb Low Pos | 108 | 14.68 | 0.66 | 4.47 | 0.00 | 0.00 | 0.00 | 0.00 | 0.00 | 0.00 | 0.30 | 2.02 | 0.83 | 5.64 |
| Calb Mod Pos | 107 | 14.37 | 0.66 | 4.58 | 0.14 | 0.99 | 0.00 | 0.00 | 0.00 | 0.00 | 0.28 | 1.98 | 0.81 | 5.64 |
| Cglab Low Pos | 106 | 21.36 | 0.84 | 3.94 | 0.18 | 0.84 | 0.00 | 0.00 | 0.00 | 0.00 | 0.62 | 2.89 | 1.26 | 5.88 |
| Cglab Mod Pos | 107 | 20.54 | 0.99 | 4.83 | 0.30 | 1.46 | 0.00 | 0.00 | 0.00 | 0.00 | 0.48 | 2.34 | 1.37 | 6.68 |
| Tvag Low Pos | 108 | 24.32 | 1.16 | 4.77 | 0.00 | 0.00 | 0.00 | 0.00 | 0.00 | 0.00 | 0.60 | 2.48 | 1.59 | 6.54 |
| Tvag Mod Pos | 107 | 23.09 | 1.18 | 5.13 | 0.00 | 0.00 | 0.00 | 0.00 | 0.00 | 0.00 | 0.56 | 2.41 | 1.56 | 6.77 |

Table 22: Signal Variability of the Aptima CV/TV Assay by Analyte-positive Panel Member

Calb = C. albicans, Cglab = C. glabrata, CV = coefficient of variation, Mod = moderate, Pos = standard deviation, TTime = emergence time of a signal (above a specific threshold), Tvag = T. vaginalis

Note: If variability from a factor was numerically negative, SD and CV are shown as 0.00.

1 The assay reports TTime for each assay analyte separately; the mean and signal variability reported are for the analyte(s) present in each panel member.

For Aptima CV/TV assay controls and positive calibrators, the total %CV values ranged

from 4.2.9% to 6.89%; total SD values were ≤1.36 (see Tables 23 and 24, respectively).

| | | | Between
Sites | | Between
Operators | | Between
Days | | Within
Runs | | Total | |
|---------------------------|----|---------------|------------------|--------|----------------------|--------|-----------------|--------|----------------|--------|-------|--------|
| Control | N | Mean
TTime | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) |
| Candida species
group- | 36 | 13.73 | 0.38 | 2.79 | 0.20 | 1.46 | 0.00 | 0.00 | 0.40 | 2.92 | 0.59 | 4.29 |
| C. glabrata- | 36 | 19.78 | 0.09 | 0.47 | 0.68 | 3.44 | 0.00 | 0.00 | 1.18 | 5.94 | 1.36 | 6.89 |
| T. vaginalis | 36 | 17.44 | 0.43 | 2.44 | 0.34 | 1.93 | 0.00 | 0.00 | 0.79 | 4.56 | 0.96 | 5.52 |

Table 23: Signal Variability of the Aptima CV/TV Assay Positive Controls

CV = coefficient of variation, SD = standard deviation, TTime = emergence time of a signal (above a specific threshold)

Note: If variability from a factor was numerically negative, SD and CV are shown as 0.00.

Calibrator Lot	Replicates	Mean RLU	Standard Deviation	%CV
CV/TV Positive Calibrator Lot 021770	480	1,344,224	68,097	5.1

	Between Sites		Between Operators		Between Days		Between Runs		Within Runs		Total
Analyte	N	Mean TTime	SD	CV (%)	SD	CV (%)	SD	CV (%)	SD	CV (%)	SD	CV (%)	SD	CV (%)
Candida species group	101	14.76	0.68	4.64	0.21	1.42	0.00	0.00	0.40	2.72	0.20	1.38	0.85	5.73

CV = coefficient of variation, SD = standard deviation, TTime = emergence time of a signal (above a specific threshold)

Note: If variability from a factor was numerically negative, SD and CV are shown as 0.00.

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VIII. CONCLUSIONS

The analytical and clinical study results demonstrate that the Aptima CV/TV assay on the Panther system performs comparably to the predicate device that is currently marketed for the same intended use. Hardware and software verification and validation demonstrate that the Aptima CV/TV assay on the Panther system will perform as intended.