(79 days)
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.
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. The 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.
Here's a breakdown of the acceptance criteria and the study proving the device meets them, based on the provided text:
Acceptance Criteria and Reported Device Performance
The document doesn't explicitly state "acceptance criteria" for clinical performance in a table format with specific numerical thresholds the device must meet. Instead, it presents the achieved performance metrics (sensitivity, specificity, PPV, NPV) from the clinical study, implying these demonstrated statistics were considered acceptable for clearance. For analytical studies, acceptance is demonstrated by successfully meeting the goals of the study (e.g., all strains detected for inclusivity, no cross-reactivity beyond a certain concentration).
Therefore, for the clinical performance, the reported performance is the demonstration of meeting implied acceptance.
Note: For a medical device, acceptance criteria are typically predefined thresholds in a study protocol that the device performance must surpass. Since these aren't explicitly stated as hard "criteria" with target numbers, I'll present the reported device performance from the clinical study tables.
| Metric | Candida species group (Patient-collected) | Candida species group (Clinician-collected) | C. glabrata (Patient-collected) | C. glabrata (Clinician-collected) | T. vaginalis (Patient-collected) | T. vaginalis (Clinician-collected) |
|---|---|---|---|---|---|---|
| Sensitivity (95% CI) | 92.9 (90.0-95.0) | 91.7 (88.7-94.0) | 86.2 (75.1-92.8) | 84.7 (73.5-91.8) | 97.1 (92.9-98.9) | 96.5 (92.0-98.5) |
| Specificity (95% CI) | 91.0 (89.1-92.6) | 94.9 (93.4-96.1) | 98.7 (98.0-99.2) | 99.1 (98.4-99.5) | 98.9 (98.2-99.4) | 95.1 (93.8-96.2) |
| PPV (95% CI) | 80.5 (77.4-83.4) | 87.8 (84.8-90.4) | 73.5 (63.7-82.2) | 79.4 (69.4-87.5) | 90.7 (85.5-94.5) | 68.5 (63.2-73.8) |
| NPV (95% CI) | 97.0 (95.8-97.9) | 96.6 (95.5-97.6) | 99.4 (99.0-99.7) | 99.4 (98.9-99.7) | 99.7 (99.2-99.9) | 99.6 (99.1-99.9) |
For Analytical Studies, acceptance was indicated by:
- Analytical Sensitivity (LoD): Predicted detection limits established through probit analysis (Table 6).
- Analytical Inclusivity: All Candida strains and 8/9 T. vaginalis strains detected at 3X LoD, with one T. vaginalis at 4X LoD.
- Cross-Reactivity and Microbial Interference: No cross-reactivity or microbial interference observed for 64 organisms and human cell lines at specified concentrations (Table 7), with minor exceptions noted for Candida famata, Tioconazole, Vaginal Moisturizing Gel, and Glacial Acetic Acid at higher concentrations than the tested limits.
- Within Laboratory Precision: 100% agreement with expected positivity for all positive and negative panel members (Table 9). Signal variability (CV%) for TTime was reported and considered acceptable (Table 10).
- Co-Infection: 100% detection for both targets present in co-infection panels (Table 11), with further testing confirming low C. glabrata and high T. vaginalis detection.
- Reproducibility: 100% agreement with expected results for all panel members across three sites (Table 21). Total %CV values for TTime ranged from 5.64% to 6.77% across organisms/panel members (Table 22), and from 4.29% to 6.89% for controls (Table 23).
Study Details
-
Sample Size Used for the Test Set and Data Provenance:
- Total Subjects Enrolled: 1519 symptomatic women.
- Evaluable Subjects: 1496.
- Sample Types: Clinician-collected and patient-collected vaginal swab specimens.
- Data Provenance:
- Country of Origin: Geographically diverse US clinical sites (21 sites).
- Retrospective/Prospective: Prospectively-collected.
-
Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of Those Experts:
- The document does not explicitly state the number of experts used to establish the ground truth.
- Qualifications of Experts: Not specified. The reference methods are described (Candida culture, PCR/bi-directional sequencing, FDA-cleared molecular TV assay, FDA-cleared culture-based TV test), but the individuals interpreting these results to form the ground truth are not detailed.
-
Adjudication Method for the Test Set:
- The document describes how the reference results for Candida and T. vaginalis were established:
- Candida species group and C. glabrata: 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.
- T. vaginalis: A patient infected status (PIS) determined for each subject based on results from an FDA-cleared molecular TV assay and an FDA-cleared culture-based TV test.
- This indicates a composite reference standard approach (multiple tests combined) rather than a simple 2+1 or 3+1 expert adjudication for diagnostic imaging, which is not applicable here.
- The document describes how the reference results for Candida and T. vaginalis were established:
-
If a Multi Reader Multi Case (MRMC) Comparative Effectiveness Study was Done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance:
- No, an MRMC comparative effectiveness study was not done. This refers to a medical device for in vitro diagnostic testing (nucleic acid amplification test) rather than an AI-assisted diagnostic imaging device/software the question implies. The study evaluates the performance of the assay itself against established reference methods.
-
If a Standalone (i.e., algorithm only without human-in-the loop performance) was done:
- Yes, this was a standalone performance study. The Aptima CV/TV assay is an in vitro diagnostic test performed on an automated Panther system. Its performance (sensitivity, specificity) is evaluated directly against reference methods without human interpretation of the assay's output as an "assistance" to a physician. 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." This is a completely automated interpretation by the device.
-
The Type of Ground Truth Used:
- Composite Reference Standard / Expert Consensus (implied by combining tests):
- For Candida species group and C. glabrata: Ground truth was established by combining Candida growth in culture and PCR/bi-directional sequencing.
- For T. vaginalis: Ground truth (Patient Infected Status - PIS) was established using an FDA-cleared molecular TV assay and an FDA-cleared culture-based TV test.
- Composite Reference Standard / Expert Consensus (implied by combining tests):
-
The Sample Size for the Training Set:
- The document does not provide information on a separate training set or its sample size. This is common for IVD assays where "training" refers more to assay development and optimization rather than machine learning model training on a specific clinical dataset. The clinical performance study described serves as the validation dataset.
-
How the Ground Truth for the Training Set Was Established:
- Since no separate training set is detailed, information on how its ground truth was established is not provided. The ground truth for the clinical validation described above was established using composite reference standards, as per point 6.
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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 presentationof vaginitis, trichomoniasis, orvulvovaginitis | Same |
| Specimen Types | Vaginal swabs in patients who aresymptomatic for vaginitis,trichomoniasis, or vulvovaginitis | Same |
| Assay Controls | Incorporates an Internal Control inevery test. Uses external positiveand 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/TechnologyPrinciple 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 vaginalisAtopobium vaginaeBacterial Vaginosis Associated Bacteria-2 (BVAB-2)Megasphaera-1• Candida spp. ( C. albicans, C. tropicalis, C. parapsilosis, C. dubliniensis )• Candida glabrata• Candida krusei• Trichomonas vaginalisThe 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 vaginalisThe 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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| BetweenDays | BetweenInstruments | BetweenOperators | BetweenLots | BetweenRuns | WithinRun | Total | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| PanelDescription | N | MeanTTime | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) |
| C. albicansLow 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. glabrataLow 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. vaginalisLow 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. albicansModerate 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. glabrataModerate 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. vaginalisModerate 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
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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
| SpecimenType | 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
| SpecimenType | 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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| SpecimenType | 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 | AptimaC. glabrataPositive | AptimaC. glabrataNegative | 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 / AfricanAmerican | 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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| BetweenSites | BetweenOperators | BetweenDays | BetweenRuns | WithinRuns | Total | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Panel Description | N | MeanTTime1 | 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).
| BetweenSites | BetweenOperators | BetweenDays | WithinRuns | Total | ||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Control | N | MeanTTime | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) |
| Candida speciesgroup- | 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.
§ 866.3975 Device that detects nucleic acid sequences from microorganisms associated with vaginitis and bacterial vaginosis.
(a)
Identification. A device that detects nucleic acid sequences from microorganisms associated with vaginitis and bacterial vaginosis is a qualitative in vitro diagnostic device intended for the detection of microbial nucleic acid sequences in vaginal specimens collected from patients with signs and symptoms of vaginitis or bacterial vaginosis. This device is intended to aid in the diagnosis of vaginitis or bacterial vaginosis when used in conjunction with clinical signs and symptoms and other laboratory findings.(b)
Classification. Class II (special controls). The special controls for this device are:(1) Design verification and validation must include:
(i) Documentation with a detailed device description of device components; ancillary reagents required but not provided; and explanation of the methodology including primer/probe sequence, design, and rationale for sequence selection.
(ii) Documentation with information that demonstrates the performance characteristics of the device, including:
(A) Limit of Detection;
(B) Precision (reproductivity);
(C) Analytical specificity;
(D) Analytical reactivity (inclusivity);
(E) Specimen stability; and
(F) Effects of interfering substances.
(iii) Detailed documentation from a prospective clinical study. As appropriate to the intended use, the prospective clinical study must be performed on an appropriate study population, including women of various ages and ethnicities. The prospective clinical study must compare the device performance to results obtained from well-accepted comparator methods.
(iv) Detailed documentation for device software, including software applications and hardware-based devices that incorporate software.
(2) The labeling required under § 809.10(b) of this chapter must include:
(i) A detailed explanation of the interpretation of results and acceptance criteria;
(ii) For devices with an intended use that includes detection of nucleic acid sequences from bacteria associated with bacterial vaginosis, clinical performance stratified by patient demographics such as race, ethnicity, age, and pregnancy status.
(iii) For devices with an intended use that includes detection of nucleic acid sequences from bacteria associated with bacterial vaginosis, a summary of device results in an asymptomatic population with demographic characteristics appropriate to the intended use population.
(iv) For devices with an intended use that includes detection of either Candida species or bacteria associated with bacterial vaginosis, a limitation that
Candida species and bacterial compositions associated with bacterial vaginosis can be present as part of normal vaginal flora and results should be considered in conjunction with available clinical information.