Predicate Devices

Reference Devices

AI/ML (Artificial Intelligence / Machine Learning)

No
The summary describes a standard nucleic acid amplification test (NAAT) for detecting bacterial vaginosis. There is no mention of AI, ML, or any computational methods beyond basic data processing for reporting qualitative results.

Therapeutic

No.
The device is an in vitro diagnostic test intended to aid in the diagnosis of bacterial vaginosis by detecting specific bacterial ribosomal RNA; it does not provide any treatment or therapy.

Diagnostic

Yes

Explanation: The "Intended Use / Indications for Use" section explicitly states, "The assay is intended to aid in the diagnosis of BV..." and the "Device Description" section reiterates that the assay provides "qualitative results to aid in the diagnosis of bacterial vaginosis." These statements clearly indicate its purpose as a diagnostic device.

SaMD (Software as a Medical Device)

No

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

IVD (In Vitro Diagnostic)

Based on the provided information, this device is an IVD (In Vitro Diagnostic).

Here's why:

Intended Use/Indications for Use: Explicitly states "in vitro nucleic acid amplification test" and "intended to aid in the diagnosis of BV". This clearly indicates it's used outside the body to analyze samples for diagnostic purposes.
Device Description: Reinforces that it's an "in vitro nucleic acid amplification test" and describes the process of analyzing a sample (vaginal swab) to provide a qualitative result for diagnosis.
Nature of the Test: The test analyzes biological material (rRNA from bacteria) from a patient sample to provide information about a medical condition (bacterial vaginosis). This is the core function of an IVD.

The information provided aligns perfectly with the definition and purpose of an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The Aptima BV assay is an in vitro nucleic acid amplification test that utilizes real time transcription-mediated amplification (TMA) for detection and quantitation of ribosomal RNA from bacteria associated with bacterial vaginosis (BV), including Lactobacillus (L. gasseri, L. crispatus, and L. jensenii), Gardnerella vaginalis, and Atopobium vaginae. The assay reports a qualitative result for BV and does not report results for individual organisms. The assay is intended to aid in the diagnosis of BV on the automated Panther system using clinician-collected and patient-collected vaginal swab specimens from females with a clinical presentation consistent with vaginitis and/or vaginosis.

Product codes

PQA, NSU, PMN

Device Description

The Aptima BV assay is an in vitro nucleic acid amplification test for the detection and quantitation of rRNA from bacteria associated with bacterial vaginosis in women with a clinical presentation consistent with vaginitis/vaginosis. The Aptima BV assay utilizes the automated Panther system to provide qualitative results to aid in the diagnosis of bacterial vaginosis.

The Aptima BV 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 cells, releases the RNA, and protects it from degradation during storage. When the Aptima BV 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 Lactobacillus group, Atopobium vaginae, Gardnerella vaginalis and IC amplification products. The Panther system software compares signal emergence times for each target organism to calibration information in order to determine the BV Positive or Negative status of each sample.

The Aptima BV assay is provided as a 100-test kit. The Aptima BV 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 BV 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 BV Calibrator and Controls kit may also be procured separately if customers need additional calibrators or controls.

The Aptima BV 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 BV 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.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Vaginal

Indicated Patient Age Range

Determined from enrollment in clinical studies:
Symptomatic: 14-75 years
Asymptomatic: 18-73 years

Intended User / Care Setting

Clinician-collected and patient-collected vaginal swab specimens from females with a clinical presentation consistent with vaginitis and/or vaginosis.
Clinical studies conducted at geographically diverse US private and academic family practice, obstetric-gynecologic, family planning, public health, STI, 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:

Sample Size: 1519 symptomatic women, 174 asymptomatic women, and 4 women with unknown symptom status were enrolled. 1417 symptomatic subjects and 172 asymptomatic subjects were evaluable.
Data Source: Prospectively-collected patient- and clinician-collected vaginal swab samples from women ≥14 years who were asymptomatic for or who exhibited signs and/or symptoms of vaginitis (ie, symptomatic). Collected from twenty-one participating geographically diverse US private and academic family practice, obstetric-gynecologic, family planning, public health, STI, medical group clinics, and clinical research centers.
Annotation Protocol:
- For each symptomatic subject, two vaginal swab samples (one patient-collected, one clinician-collected) were tested with the investigational Aptima BV assay.
- One clinician-collected vaginal swab sample was used for Nugent score evaluation, and modified Amsel criteria if necessary, to determine BV infection status.
- A Nugent interpretation established positive and negative BV reference status, except in cases of intermediate determinations. For intermediate Nugent interpretations, BV reference status was established using modified Amsel criteria.
- For each asymptomatic subject, one (1) clinician-collected vaginal swab sample was collected and tested with the investigational Aptima BV assay.

Reproducibility Study:

Sample Size: Reproducibility was evaluated at three US sites using seven panel members. Testing was performed for at least six days at each site. Each run had three replicates of each panel member.
Data Source: Panel members were made using a simulated vaginal swab matrix ('SVSM', which contains specimen transport media (STM) spiked with simulated vaginal fluid) negative for Lactobacillus species, G. vaginalis, and A. vaginae. Six panel members contained cell lysates of at least 1 of the following organisms: L. crispatus, L. jensenii, G. vaginalis, or A. vaginae; different bacterial combinations were prepared to represent the variety of targeted BV organism combinations present in vaginal specimens. One negative panel member contained only the matrix with no added target analytes.
Annotation Protocol: Not explicitly stated, but testing was performed using one lot of assay reagents and six operators (two at each site).

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

Analytical (Non-Clinical) Studies:

Analytical Sensitivity (Limit of Detection or LoD): Determined by testing a series of panels of L. crispatus, L. gasseri, L. jensenii, G. vaginalis, or A. vaginae cell lysates diluted into simulated vaginal swab matrix (SVSM). Minimum of 20 replicates of each panel member tested with two reagent lots (minimum 40 replicates per panel member). Probit analysis used to calculate predicted detection limits.
- Predicted Detection Limits (95%): A. vaginae (2901 CFU/mL), G. vaginalis (rata CFU/mL), L. crispatus (143 CFU/mL), L. gasseri (2,207 CFU/mL), L. jensenii (10 CFU/mL).
- Predicted BV Positivity Limits (C95) for A. vaginae and G. vaginalis are approximately 5.10 log CFU/mL and 4.86 log CFU/mL, respectively.
Analytical Inclusivity: Five strains of each target organism tested at 3X C95 for G. vaginalis and A. vaginae, and 3X LoD for Lactobacillus species in SVSM.
- Aptima BV Assay was BV positive for all five strains of G. vaginalis and A. vaginae at 3X C95.
- All five strains of L. crispatus and L. gasseri were detected at 3X LoD.
- Three of five strains of L. jensenii were detected at 3X LoD, and the remaining two strains at 10X LoD.
Cross-Reactivity and Microbial Interference: Evaluated in presence of non-targeted organisms using a panel of 62 organisms in SVSM, with or without L. crispatus (3X LoD), G. vaginalis (3X C95), or A. vaginae (3X C95).
- No cross-reactivity or microbial interference observed for any of the 62 organisms tested at specified concentrations, except for Lactobacillus acidophilus which affects BV positivity at 1x104 CFU/mL or higher.
Interference: Potentially interfering substances tested in SVSM. Sensitivity evaluated for L. crispatus (3X LoD), G. vaginalis (3X C95), and A. vaginae (3X C95). Specificity evaluated with negative panels.
- No interference observed for listed exogenous and endogenous substances at specified concentrations, except for Mucus at ≥2% V/V (interference observed, not at 1.5% V/V), Tioconazole 6.5% Ointment at 5% W/V (interference observed, not at 1% W/V), and Vaginal Moisturizing Gel at ≥0.5% W/V (interference observed, not at 0.4% W/V).
Within Laboratory Precision: Evaluated on three Panther systems at one site by three operators across 21 days and three reagent lots. Panel of 11 members (SVSM with and without organisms).
- BV percent positive results: SVSM (0%), L. crispatus, A. vaginae BV Negative (0%), L. crispatus, G. vaginalis BV High Negative (45.2%), L. crispatus, G. vaginalis, A. vaginae BV High Negative (79.4%), G. vaginalis BV Low Positive (100%), A. vaginae BV Low Positive (100%), L. jensenii, A. vaginae BV Low Positive (100%), G. vaginalis, A. vaginae BV Low Positive (100%), L. crispatus, G. vaginalis, A. vaginae BV Low Positive (100%), G. vaginalis BV Mod Positive (100%), A. vaginae BV Mod Positive (100%).
- Signal variability (TTime) for Lactobacillus, G. vaginalis, and A. vaginae components was calculated, showing overall CVs ranging from 3.30% to 5.74%.

Clinical Studies:

Clinical Performance Study: Multicenter study using prospectively-collected patient- and clinician-collected vaginal swab samples. (Detailed sample size and data source in "Description of the test set").
- Key Results (Symptomatic Subjects):
  - Patient-collected: Sensitivity 97.3% (95% CI: 95.8-98.2), Specificity 85.8% (95% CI: 83.1-88.2), PPV 87.0% (95% CI: 84.8-88.9), NPV 97.0% (95% CI: 95.5-98.1). Prevalence 49.3%.
  - Clinician-collected: Sensitivity 95.0% (95% CI: 93.1-96.4), Specificity 89.6% (95% CI: 87.1-91.6), PPV 89.8% (95% CI: 87.7-91.7), NPV 94.8% (95% CI: 93.1-96.3). Prevalence 49.2%.
  - Performance also analyzed by race/ethnicity and clinical condition (tables provided for detailed results).
- Positivity Rates (Asymptomatic Subjects): Overall 40.7% (70/172). Race/Ethnicity breakdown provided.
Reproducibility Study: Evaluated at three US sites using seven panel members. (Detailed sample size and data source in "Description of the test set").
- Key Results:
  - Agreement with expected results was 100% (95% CI: 96.6-100) for all panel members containing L. crispatus, L. jensenii, G. vaginalis, or A. vaginae, and for true negative controls.
  - Total %CV values for organisms/panel members ranged from 4.21% to 4.76%; total SD values were ≤1.12.
  - Total %CV values for Aptima BV assay controls and positive calibrators ranged from 4.47% to 5.36%; total SD values were ≤1.11.
  - "between sites," "between operators," and "between runs" factors were the largest contributors to total variability for most panel members.

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

Clinical Performance Study (Symptomatic Subjects):

Patient-collected:
- Sensitivity: 97.3% (95% CI: 95.8-98.2)
- Specificity: 85.8% (95% CI: 83.1-88.2)
- PPV: 87.0% (95% CI: 84.8-88.9)
- NPV: 97.0% (95% CI: 95.5-98.1)
- Prevalence: 49.3%
Clinician-collected:
- Sensitivity: 95.0% (95% CI: 93.1-96.4)
- Specificity: 89.6% (95% CI: 87.1-91.6)
- PPV: 89.8% (95% CI: 87.7-91.7)
- NPV: 94.8% (95% CI: 93.1-96.3)
- Prevalence: 49.2%

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 logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

May 23, 2019

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

Re: K190452

Trade/Device Name: Aptima BV Assay, Aptima BV Controls Kit, Aptima BV Calibrator Kit 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, PMN Dated: February 22, 2019 Received: February 25, 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 requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part

1

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 (QS) 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

2

510(k) SUMMARY Aptima BV 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
Fax: (858) 410-5557 |

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

Date Prepared: February 19, 2019

II. DEVICE

Proprietary Name of Device:	Aptima BV 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, PMN

III. PREDICATE DEVICE

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

IV. DEVICE DESCRIPTION

The Aptima BV assay is an in vitro nucleic acid amplification test for the detection and quantitation of rRNA from bacteria associated with bacterial vaginosis in women with a clinical presentation consistent with vaginitis/vaginosis. The Aptima BV assay utilizes the automated Panther system to provide qualitative results to aid in the diagnosis of bacterial vaginosis.

3

Principles of the Procedure

The Aptima BV 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 cells, releases the RNA, and protects it from degradation during storage. When the Aptima BV 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 Lactobacillus group, Atopobium vaginae, Gardnerella vaginalis and IC amplification products. The Panther system software compares signal emergence times for each target organism to calibration information in order to determine the BV Positive or Negative status of each sample.

4

Assay Components

The Aptima BV assay is provided as a 100-test kit. The Aptima BV 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 BV 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 BV 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 BV 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
	Enzyme Reagent
	Promoter Reagent
	Internal Control
2	Amplification Reconstitution Solution
	Enzyme Reconstitution Solution
	Promoter Reconstitution Solution
	Target Capture Reagent
3	Positive Calibrator
4	Negative Control
	Positive Control

Table 1: Reagents Required to Perform the Aptima BV Assay

Table 2: Ancillary and Collection Kits Required to Perform the Aptima BV Assay
Aptima Assay Fluids Kit
Aptima Multitest Swab Specimen Collection Kit

Instrumentation

The Aptima BV 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 BV 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.

5

V. INDICATIONS FOR USE

The Aptima BV assay is an in vitro nucleic acid amplification test that utilizes real time transcription-mediated amplification (TMA) for detection and quantitation of ribosomal RNA from bacteria associated with bacterial vaginosis (BV), including Lactobacillus (L. gasseri, L. crispatus, and L. jensenii), Gardnerella vaginalis, and Atopobium vaginae. The assay reports a qualitative result for BV and does not report results for individual organisms. The assay is intended to aid in the diagnosis of BV on the automated Panther system using clinician-collected and patient-collected vaginal swab specimens from females with a clinical presentation consistent with vaginitis and/or vaginosis.

VI. COMPARISON OF TECHNOLOGICAL CHARACTERISTICS WITH THE PREDICATE DEVICE

A comparison of the Aptima BV 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 BV Assay
(Subject Device) |
|--------------------|-----------------------------------------------------------------------------------------------------|-------------------------------------|
| Patient Population | Women with a clinical presentation
of vaginitis/vaginosis | Same |
| Specimen Types | Vaginal swabs in patients who are
symptomatic for vaginitis/vaginosis | 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 BV Assay
(Subject Device) |
|--------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Organisms Detected | Lactobacillus (L. crispatus, and L.
jensenii), Lactobacillus (L. gasseri,
L. crispatus, and L. jensenii),
Atopobium vaginae, Bacterial
Vaginosis Associated Bacteria-2
(BVAB-2), Megasphaera-1,
Candida (C. albicans, C.
tropicalis, C. parapsilosis, C.) | Lactobacillus (L. gasseri, L.
crispatus, and L. jensenii),
Gardnerella vaginalis, and
Atopobium vaginae |
| Item | BD MAX Vaginal Panel
(DEN160001) | Aptima BV Assay
(Subject Device) |
| | dubliniensis), Candida glabrata,
Candida krusei, 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 | ribosomal RNA |
| Assay Calibrators | None | Positive Calibrator |
| Intended Use | The BD MAX Vaginal Panel
performed on the BD MAX 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. | The Aptima BV assay is an in vitro
nucleic acid amplification test that
utilizes real time transcription-
mediated amplification (TMA) for
detection and quantitation of
ribosomal RNA from bacteria
associated with bacterial vaginosis
(BV), including Lactobacillus ( L.
gasseri, L. crispatus, and L.
jensenii ), Gardnerella vaginalis ,
and Atopobium vaginae . The assay
reports a qualitative result for BV
and does not report results for
individual organisms. The assay is
intended to aid in the diagnosis of
BV on the automated Panther
system using clinician-collected
and patient-collected vaginal swab
specimens from females with a
clinical presentation consistent
with vaginitis and/or vaginosis. |
| Item | BD MAX Vaginal Panel
(DEN160001) | Aptima BV Assay
(Subject Device) |
| | 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
trichomoniasis. | |

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7

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 BV assay on the Panther system.

Analytical Sensitivity

The analytical sensitivity (Limit of Detection or LoD) and BV positivity limits of the Aptima BV assay were determined by testing a series of panels consisting of L. crispatus, L. gasseri, L. jensenii, G. vaginalis, or A. vaginae cell lysates diluted into simulated vaginal swab matrix (SVSM). A minimum of 20 replicates of each panel member were tested with each of two reagent lots for a minimum of 40 replicates per panel member. The predicted detection limits for each organism calculated using Probit analysis are shown in Table 6.

8

| Organism | Predicted Detection
Limit | CFU/mL |
|--------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------|
| A. vaginae | 95% | 2901 |
| G. vaginalis | 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 | રતા |
| L. crispatus | 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 લોકોનો મુખ્ય વ્યવસાય ખેતી, ખેતમજૂરી | 143 |
| L. gasseri | 95% | 2,207 |
| L. jensenii | 95% | 10 |

Table 6: Limit of Detection of the Aptima BV Assay

1 Predicted BV Positivity Limits (C95) for A. vaginae and G. vaginalis in the Aptima BV assay are approximately 5.10 log CFU/mL and 4.86 log CFU/mL, respectively.

Analytical Inclusivity

Five strains of each target organism were tested using lysate targeting 3X C95 for G. vaginalis and A. vaginae, and 3X LoD for Lactobacillus species (L. crispatus, L. gasseri, and L. jensenii) in SVSM. The Aptima BV Assay was BV positive for all five strains of G. vaginalis and A. vaginae at 3X C95. All five strains of L. crispatus and L. gasseri were detected at 3X LoD. Three of the five strains of L. jensenii were detected at 3X LoD, and the remaining two strains at 10X LoD .

Cross-Reactivity and Microbial Interference

Cross-reactivity and microbial interference with the Aptima BV assay was evaluated in the presence of non-targeted organisms. A panel consisting of 62 organisms (Table 7) was tested in SVSM in the absence or in the presence of L. crispatus at 3X LoD, G. vaginalis at 3X C95, or A. vaginae at 3X C95. No cross-reactivity or microbial interference was observed for any of the 62 organisms tested in the Aptima BV assay at the following concentrations.

9

Microorganism	Concentration	Microorganism	Concentration
Acinetobacter lwoffii	1x106 CFU/mL	Herpes simplex virus I	1x104 TCID50/mL
Actinomyces israelii	1x106 CFU/mL	Herpes simplex virus II	1x104 TCID50/mL
Alcaligenes faecalis	1x106 CFU/mL	HIV	1x105 copies/mL
Atopobium minutum	1x106 CFU/mL	Klebsiella pneumoniae	1x106 CFU/mL
Atopobium parvulum	1x106 CFU/mL	Lactobacillus acidophilus	1x103 CFU/mL2
Atopobium rimae	1x106 CFU/mL	Lactobacillus iners	1x106 CFU/mL
Bacteroides fragilis	1x106 CFU/mL	Lactobacillus mucosae	1x106 CFU/mL
Bifidobacterium adolescentis	1x106 CFU/mL	Leptotrichia buccalis	1x106 CFU/mL
Bifidobacterium breve	1x106 CFU/mL	Listeria monocytogenes	1x106 CFU/mL
BVAB-11	1x106 copies/mL	Megasphaera Type 11	1x106 copies/mL
BVAB-21	1x106 copies/mL	Mobiluncus curtisii	1x106 CFU/mL
Campylobacter jejuni	1x106 CFU/mL	Mycoplasma genitalium	1x106 CFU/mL
Candida albicans	1x106 CFU/mL	Mycoplasma hominis	1x106 CFU/mL
Candida dubliniensis	1x106 CFU/mL	Neisseria gonorrhoeae	1x106 CFU/mL
Candida glabrata	1x106 CFU/mL	Pentatrichomonas hominis	1x105 cells/mL
Candida krusei	1x106 CFU/mL	Peptostreptococcus magnus	1x106 CFU/mL
Candida lusitaniae	1x106 CFU/mL	Pichia fermentans	1x106 CFU/mL
Candida orthopsilosis	1x106 CFU/mL	Prevotella bivia	1x106 CFU/mL
Candida parapsilosis	1x106 CFU/mL	Propionibacterium acnes	1x106 CFU/mL
Candida tropicalis	1x106 CFU/mL	Proteus vulgaris	1x106 CFU/mL
Chlamydia trachomatis	1x106 IFU/mL	SiHa cells	1x104 cells/mL
Clostridium difficile	1x106 CFU/mL	Sneathia amnii	1x106 CFU/mL
Corynebacterium genitalium	1x106 CFU/mL	Staphylococcus aureus	1x106 CFU/mL
Cryptococcus neoformans	1x106 CFU/mL	Staphylococcus epidermidis	1x106 CFU/mL
Eggerthella lenta	1x106 CFU/mL	Streptococcus agalactiae	1x106 CFU/mL
Enterobacter cloacae	1x106 CFU/mL	Streptococcus pyogenes	1x106 CFU/mL
Enterococcus faecalis	1x106 CFU/mL	Treponema pallidum1	1x106 copies/mL
Escherichia coli	1x106 CFU/mL	Trichomonas tenax	1x105 cells/mL
Fusobacterium nucleatum	1x106 CFU/mL	Trichomonas vaginalis	1x105 cells/mL
Haemophilus ducreyi	1x106 CFU/mL	Ureaplasma parvum	1x106 CFU/mL
HeLa cells	1x104 cells/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 Lactobacillus acidophilus affects BV positivity at 1x104 CFU/mL or higher.

10

Interference

Potentially interfering substances were tested in the Aptima BV assay. Panels were built in SVSM and evaluated for potential effects on assay sensitivity and specificity. Sensitivity performance was evaluated separately for L. crispatus by spiking lysate at 3X LoD, and for G. vaginalis and A. vaginae by spiking lysate at 3X C95. 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
Mucus2	1.5% V/V
Seminal Fluid	5% V/V
Contraceptive Foam	5% W/V
Contraceptive Film	5% W/V
Tioconazole3	1% 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 Gel4	0.4% W/V
Lubricant	5% V/V
Spermicide	5% W/V
Anti-fungal	5% W/V
Deodorant/Spray	5% W/V
Glacial Acetic Acid	5% 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 Interference was observed with Mucus at ≥2% V/V and not observed at 1.5% V/V.

3 Interference was observed with Tioconazole 6.5% Ointment at 5% W/V and not observed at 1% W/V.

4 Interference was observed with Vaginal Moisturizing Gel at ≥0.5% W/V and not observed at 0.4% W/V.

Within Laboratory Precision

Within Lab Precision was evaluated on three Panther systems at one site. Three operators performed testing across 21 days and three reagent lots. Each operator performed two runs

11

per day using an 11 member panel. Each run consisted of three replicates of each panel member. The panel members were made using SVSM negative for Lactobacillus species. G. vaginalis, and A. vaginae. Ten panel members contained cell lysates of at least 1 of the following organisms: L. crispatus, L. jensenii, G. vaginalis, or A. vaginae; different bacterial combinations were prepared to represent the variety of targeted BV organism combinations present in vaginal specimens. Ten panel members targeted BV Negative (50 | 172 (12.1) | 52 (30.2) |
| Race/Ethnicity, n (%) | | |
| Asian | 67 (4.7) | 5 (2.9) |
| Black or African American | 731 (51.6) | 75 (43.6) |
| White (Hispanic or Latino) | 248 (17.5) | 41 (23.8) |
| White (Not Hispanic or Latino) | 307 (21.7) | 44 (25.6) |
| Other¹ | 64 (4.5) | 7 (4.1) |
| ¹Includes patient-reported other, mixed, and unknown races | | |

Table 13: Summary of Demographics of Evaluable Subjects in the Aptima BV Assay Evaluation

Performance characteristics for detection of BV infection for patient-collected and cliniciancollected vaginal swab samples from symptomatic subjects were calculated overall (see Table 14), by race/ethnicity (see Table 15), and by clinical condition (see Table 16).

16

| Specimen
Type | N | TP | FP | TN | FN | Prevalence1
(95% CI)2 | Sensitivity
(95% CI)2 | Specificity
(95% CI)2 | PPV
(95% CI)3 | NPV
(95% CI)3 |
|-------------------------|------|-----|------|-----|-----|--------------------------|--------------------------|--------------------------|------------------|------------------|
| Patient-
collected | 1405 | 673 | 1014 | 612 | 195 | 49.3 | 97.3 (95.8-98.2) | 85.8 (83.1-88.2) | 87.0 (84.8-88.9) | 97.0 (95.5-98.1) |
| Clinician-
collected | 1413 | 660 | 756 | 643 | 357 | 49.2 | 95.0 (93.1-96.4) | 89.6 (87.1-91.6) | 89.8 (87.7-91.7) | 94.8 (93.1-96.3) |

Table 14: Aptima BV Assay Performance Relative to BV Infection Status in Symptomatic Subjects

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

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

4 Of the 101 false positive results, 55 subjects were Nugent interion status determined by Amsel criteria, and 9 were positive by Amsel.

5 Of the 19 false negative results, 6 subjects were Nigent internediates and had BV infection status determined by Amsel.

6 Of the 75 false positive results, 46 subjects were Nued BV infection status determined by Amsel criteria, and 6 were positive by Amsel. 7 Of the 35 false negative results, 10 subjects were Nugent internetiates and had BV infection status determined by Amsel criteria, and 15 were negative by Amsel.

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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 | 65 | 19 | 6 | 39 | 1 | 30.8 | 95.0 (76.4-99.1) | 86.7 (73.8-93.7) | 76.0 (61.6-88.7) | 97.5 (89.3-99.9) |
| Black / African American | 727 | 434 | 43 | 239 | 11 | 61.2 | 97.5 (95.6-98.6) | 84.8 (80.1-88.5) | 91.0 (88.6-93.1) | 95.6 (92.6-97.7) |
| White (Hispanic/Latino) | 246 | 112 | 22 | 111 | 1 | 45.9 | 99.1 (95.2-99.8) | 83.5 (76.2-88.8) | 83.6 (78.0-88.6) | 99.1 (95.6- 100) |
| White (Not Hispanic/Latino) | 303 | 81 | 27 | 189 | 6 | 28.7 | 93.1 (85.8-96.8) | 87.5 (82.4-91.3) | 75.0 (68.1-81.5) | 96.9 (94.0-98.8) |
| Other4 | 64 | 27 | 3 | 34 | 0 | 42.2 | 100 (87.5-100) | 91.9 (78.7-97.2) | 90.0 (76.9-97.7) | 100 (91.3- 100) |
| Clinician-collected | | | | | | | | | | |
| Asian | 67 | 20 | 4 | 42 | 1 | 31.3 | 95.2 (77.3-99.2) | 91.3 (79.7-96.6) | 83.3 (68.2-94.3) | 97.7 (89.8-99.9) |
| Black / African American | 729 | 425 | 31 | 253 | 20 | 61.0 | 95.5 (93.2-97.1) | 89.1 (84.9-92.2) | 93.2 (90.9-95.2) | 92.7 (89.4-95.2) |
| White (Hispanic/Latino) | 247 | 110 | 18 | 115 | 4 | 46.2 | 96.5 (91.3-98.6) | 86.5 (79.6-91.3) | 85.9 (80.3-90.8) | 96.6 (92.3-99.0) |
| White (Not Hispanic/Latino) | 306 | 78 | 18 | 200 | 10 | 28.8 | 88.6 (80.3-93.7) | 91.7 (87.3-94.7) | 81.3 (73.9-87.5) | 95.2 (92.1-97.5) |
| Other4 | 64 | 27 | 4 | 33 | 0 | 42.2 | 100 (87.5-100) | 89.2 (75.3-95.7) | 87.1 (74.2-96.0) | 100 (91.1- 100) |

Table 15: Aptima BV Assay Performance Relative to BV Infection Status in Symptomatic Subjects, by Race/Ethnicity

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

Study prevalence reported. 35core 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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Clinical Condition	N	TP	FP	TN	FN	Prev1 (%)	Sensitivity (95% CI)2	Specificity (95% CI)2	PPV (95% CI)3	NPV (95% CI)3
Patient-collected
Use of antibiotics	3	1	0	2	0	33.3	100 (20.7-100)	100 (34.2-100)	100 ( 7.8- 100)	100 (45.1- 100)
Use of antifungals	8	2	0	6	0	25.0	100 (34.2-100)	100 (61.0-100)	100 (33.3- 100)	100 (77.4- 100)
Use of estrogen therapy	2	0	0	2	0	0.0	NC	100 (34.2-100)	NC	100 ( NC )
Recurrent symptoms of vaginitis in the last 12 months	828	405	62	353	8	49.9	98.1 (96.2-99.0)	85.1 (81.3-88.2)	86.7 (83.9-89.3)	97.8 (95.9-99.0)
Unprotected intercourse in the last 24 hours	94	53	10	30	1	57.4	98.1 (90.2-99.7)	75.0 (59.8-85.8)	84.1 (76.5-91.0)	96.8 (85.6-99.9)
Pregnant	20	9	1	10	0	45.0	100 (70.1-100)	90.9 (62.3-98.4)	90.0 (66.5-99.7)	100 (77.9- 100)
With Menses	109	52	9	48	0	47.7	100 (93.1-100)	84.2 (72.6-91.5)	85.2 (76.6-92.4)	100 (93.6- 100)
Without Menses	1175	579	85	496	15	50.6	97.5 (95.9-98.5)	85.4 (82.3-88.0)	87.2 (84.9-89.3)	97.1 (95.4-98.3)
Post-menopausal	121	42	7	68	4	38.0	91.3 (79.7-96.6)	90.7 (82.0-95.4)	85.7 (75.5-93.2)	94.4 (88.1-98.3)
Clinician-collected
Use of antibiotics	3	1	0	2	0	33.3	100 (20.7-100)	100 (34.2-100)	100 ( 7.8- 100)	100 (45.1- 100)
Use of antifungals	8	2	0	6	0	25.0	100 (34.2-100)	100 (61.0-100)	100 (33.3- 100)	100 (77.4- 100)
Use of estrogen therapy	2	0	0	2	0	0.0	NC	100 (34.2-100)	NC	100 ( NC )
Recurrent symptoms of vaginitis in the last 12 months	832	394	47	371	20	49.8	95.2 (92.7-96.9)	88.8 (85.4-91.4)	89.3 (86.6-91.8)	94.9 (92.5-96.7)
Unprotected intercourse in the last 24 hours	94	50	6	34	4	57.4	92.6 (82.4-97.1)	85.0 (70.9-92.9)	89.3 (81.2-95.4)	89.5 (78.4-96.6)
Pregnant	20	9	0	11	0	45.0	100 (70.1-100)	100 (74.1-100)	100 (74.2- 100)	100 (78.4- 100)
With Menses	111	50	8	51	2	46.8	96.2 (87.0-98.9)	86.4 (75.5-93.0)	86.2 (77.6-93.1)	96.2 (88.7-99.5)
Without Menses	1177	569	62	520	26	50.6	95.6 (93.7-97.0)	89.3 (86.6-91.6)	90.2 (88.0-92.2)	95.2 (93.3-96.8)
Post-menopausal	125	41	5	72	7	38.4	85.4 (72.8-92.8)	93.5 (85.7-97.2)	89.1 (79.1-95.8)	91.1 (84.7-95.9)

Table 16: Aptima BV Assay Performance Relative to BV Infection Status in Symptomatic Subjects, by Clinical Condition

FN = false negative, FP = false positive, NC = not calculable, NPV = positive predictive value, PPV = positive predictive value, Prev = prevalence TP = true positive, TN = true negative

Study prevalence reported. 35core Cl - PPV 95% Cl for positive likelihod ratio; NPV 95% Cl computed from the 95% for the negative likelihood ratio.

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Positivity rates were estimated for clinician-collected vaginal swab samples from asymptomatic subjects overall and by race/ethnicity (see Table 17).

| Subjects | % Positivity
(# positive/# tested with valid results) |
|-----------------------------|----------------------------------------------------------|
| All | 40.7% (70/172) |
| Asian | 40.0% (2/5) |
| Black/African American | 52.0% (39/75) |
| White (Hispanic/Latino) | 43.9% (18/41) |
| White (Not Hispanic/Latino) | 15.9% (7/44) |
| Other | 57.1% (4/7) |

Table 17: Positivity of BV Infection as Determined by the Aptima BV Assay in Asymptomatic Subjects

Reproducibility

Aptima BV 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 Lactobacillus species, G. vaginalis, and A. vaginae. Six panel members contained cell lysates of at least 1 of the following organisms: L. crispatus, L. jensenii, G. vaginalis, or A. vaginae; different bacterial combinations were prepared to represent the variety of targeted BV organism combinations present in vaginal specimens. 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 L. crispatus, L. jensenii, G. vaginalis, or A. vaginae, as shown in Table 18.

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Panel Description	Expected Results	N	Agreement, % (95% CI)
True Neg	Negative	36/36	100 (96.6-100)
BV Neg1	Negative	36/36	100 (96.6-100)
Gvag Low Pos	Positive	36/36	100 (96.6-100)
Avag Low Pos	Positive	36/36	100 (96.6-100)
BV Low Pos1	Positive	36/36	100 (96.6-100)
Gvag Mod Pos	Positive	36/36	100 (96.6-100)
Avag MosPos	Positive	36/36	100 (96.6-100)

Table 18: Agreement of Aptima BV Assay Results With Expected Results

Avag = A. vaginae, CI = Score confidence interval, Gvag = G. vaginalis, Mod=moderate, Neg = negative, Pos = positive 1 Panel member contains 2 different organisms.

Across organisms/panel members, the total %CV values ranged from 4.21% to 4.76%; total SD values were ≤1.12. For most panel members, the "between sites," "between operators," and "between runs" factors were the largest contributors to total variability; for all other sources of variation, SD values were ≤0.18 (%CV values were ≤1.13%), as shown in Table 19.

| | | 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 (%) |
| Lcrisp BV Neg2 | 108 | 19.73 | 0.30 | 1.53 | 0.61 | 3.07 | 0.13 | 0.64 | 0.63 | 3.17 | 0.12 | 0.62 | 0.94 | 4.76 |
| Ljen Low Pos2 | 108 | 24.31 | 0.00 | 0.00 | 0.77 | 3.16 | 0.00 | 0.00 | 0.80 | 3.28 | 0.15 | 0.62 | 1.12 | 4.60 |
| Gvag Low Pos | 108 | 15.69 | 0.35 | 2.26 | 0.40 | 2.52 | 0.00 | 0.00 | 0.38 | 2.43 | 0.15 | 0.96 | 0.67 | 4.28 |
| Gvag Mod Pos | 108 | 14.33 | 0.30 | 2.07 | 0.37 | 2.58 | 0.00 | 0.00 | 0.35 | 2.41 | 0.14 | 0.98 | 0.60 | 4.21 |
| Avag BV Neg2 | 108 | 18.01 | 0.39 | 2.15 | 0.44 | 2.46 | 0.08 | 0.45 | 0.47 | 2.59 | 0.18 | 0.97 | 0.78 | 4.30 |
| Avag Low Pos | 108 | 14.95 | 0.38 | 2.52 | 0.41 | 2.75 | 0.00 | 0.00 | 0.39 | 2.61 | 0.14 | 0.93 | 0.69 | 4.64 |
| Avag Low Pos2 | 108 | 14.94 | 0.41 | 2.76 | 0.37 | 2.51 | 0.00 | 0.00 | 0.37 | 2.45 | 0.17 | 1.13 | 0.69 | 4.60 |
| Avag Mod Pos | 108 | 13.99 | 0.29 | 2.08 | 0.36 | 2.60 | 0.03 | 0.18 | 0.39 | 2.82 | 0.14 | 1.00 | 0.63 | 4.48 |

Table 19: Signal Variability of the Aptima BV Assay by Analyte-positive Panel Member

Avag = A. vaginae, CV = coefficient of vag = G. vaginalis, Lcrisp = L. crispatus, Ljen = L. jensenii, Mod = moderate, Neg = negative, Pos = positive, 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 0.0.

2 Panel member contains 2 different organisms; results are shown for only the component shown.

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.

21

For Aptima BV assay controls and positive calibrators, the total %CV values ranged from 4.47% to 5.36%; total SD values were ≤1.11 (see Tables 20 and 21, respectively).

| | | | Between
Sites | | Between
Operators | | Between
Days | | Within
Runs | | Total | |
|--------------|----|------------|------------------|--------|----------------------|--------|-----------------|--------|----------------|--------|-------|--------|
| Control | N | Mean
Ct | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) |
| Neg - Lcrisp | 36 | 20.34 | 0.24 | 1.16 | 0.74 | 3.63 | 0.00 | 0.00 | 0.71 | 3.48 | 1.05 | 5.16 |
| Pos - Gvag | 36 | 17.52 | 0.05 | 0.28 | 0.60 | 3.41 | 0.00 | 0.00 | 0.52 | 2.96 | 0.79 | 4.52 |
| Pos - Avag | 36 | 13.79 | 0.27 | 1.97 | 0.38 | 2.77 | 0.00 | 0.00 | 0.41 | 2.97 | 0.62 | 4.51 |

Table 20: Signal Variability of the Aptima BV Assav Positive Controls

Avag = A. vaginae, CV = coefficient of vag = G. vaginalis, Lcrisp = L. crispatus, Ljen = L. jensenii, Mod = moderate, Neg = negative, Pos = positive, SD = standard deviation, TTime = emergence time of a signal (above a specific threshold)

| | 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 (%) |
| Lcrisp | 108 | 20.75 | 0.00 | 0.00 | 0.78 | 3.77 | 0.00 | 0.00 | 0.78 | 3.75 | 0.13 | 0.64 | 1.11 | 5.36 |
| Gvag | 108 | 19.03 | 0.34 | 1.78 | 0.61 | 3.19 | 0.00 | 0.00 | 0.56 | 2.96 | 0.15 | 0.79 | 0.91 | 4.76 |
| Avag | 108 | 18.46 | 0.41 | 2.23 | 0.48 | 2.62 | 0.08 | 0.42 | 0.51 | 2.76 | 0.11 | 0.58 | 0.83 | 4.47 |

Table 21. Signal Variability of the Antima RV Assay Positive Calibrators

Avag = A. vaginae, CV = coefficient of variation, Gvag = G. vaginalis, Lcrispatus, SD = standard deviation, TTime = energence time of a signal (above a specific threshold)

VIII. CONCLUSIONS

The analytical and clinical study results demonstrate that the Aptima BV 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 BV assay on the Panther system will perform as intended.