(28 days)
Aptima BV Assay: The Aptima BV Assay is an in vitro nucleic acid amplification test that utilizes real time Transcription-Mediated Amplification (TMA) technology for detection and quantitation of ribosomal RNA from bacterial vaginosis (BV), including Lactobacillus (L. gasseri, L. crispatus, and L. jenseni), Gardnerella vaginalis), and Atopobium vaginae (A. 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.
Aptima CV/TV Assay: 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) technology to detect and qualitatively report results for the following organisms: Candida species group (C. albicans, C. tropicalis, C. parapsilosis, C. dubliniensis), Candida glabrata (C. glabrata), Trichomonas vaginalis (TV). The assay differentiates between C. 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 TV, the assay targets ribosomal RNA (rRNA) and differentiates the results for C. glabrata and C spp. The assy is intended to aid in the diagnosis of vulvovaginal candidiasis and trichomoniasis on the automated Panther System using clinician-collected and patientcollected vaginal swab specimens from females with a clinical presentation consistent with vagintis.
Aptima BV Assay: Like the Aptima BV assay 100 test kit, the Aptima BV assay 250 test kit 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.
Aptima CV/TV Assay: Like the Aptima CV/TV assay 100 test kit, the Aptima CV/TV assay 250 test kit is an in vitro nucleic acid amplification test for the detection and quantitation of RNA from microorganisms associated with vaginitis, trichomoniasis, and vulvovaginitis, in women with a clinical presentation consistent with vaginitis, trichomoniasis, and 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 provided document, a 510(k) summary for the Hologic Aptima BV Assay and Aptima CV/TV Assay, describes the device's technical specifications and a general statement regarding performance data for substantial equivalence. However, it does not contain the detailed acceptance criteria and a comprehensive study report with specific performance metrics (like sensitivity, specificity, PPV, NPV against a clinical gold standard) that would typically be expected for demonstrating a device meets acceptance criteria in a clinical validation context.
Specifically, the document focuses on demonstrating substantial equivalence of a new kit configuration (250 tests) to an already cleared kit configuration (100 tests) by showing similar analytical performance, particularly Limit of Detection (LoD). It does not present a de novo clinical study with pre-defined acceptance criteria for diagnostic accuracy against a true clinical ground truth.
Therefore, many of the requested items (e.g., number of experts, adjudication method, MRMC studies, training set details) are not applicable or not provided in this specific type of submission, which is for a modification to an already cleared device, not a novel device requiring full clinical validation from scratch.
Here's an attempt to answer the questions based only on the provided text:
1. A table of acceptance criteria and the reported device performance
The document does not explicitly present a table of acceptance criteria and reported device performance in terms of diagnostic accuracy (e.g., sensitivity, specificity) against a clinical reference for the 250-test kit. Instead, it asserts equivalence by confirming L.O.D. for the new kit configuration.
The primary acceptance criteria for the new 250-test kit configuration, as implied by the performance data section, is that it must meet the established Limit of Detection (LoD) of the previously cleared 100-test kit configuration.
Aptima BV Assay (250 Test Kit):
| Acceptance Criteria (LoD of 100-Test Kit) | Reported Performance (LoD Confirmation for 250-Test Kit) |
|---|---|
| L. crispatus (LC): 143 CFU/mL | LC: 143 CFU/mL |
| L. gasseri (LG): 2,207 CFU/mL | LG: 2,207 CFU/mL |
| L. jenseni (LJ): 10 CFU/mL | LJ: 10 CFU/mL |
| A. vaginae (AV) C95: 5.10 log CFU/mL | AV C95: 5.10 log CFU/mL (128,397 CFU/mL) |
| G. vaginalis (GV) C95: 4.86 log CFU/mL | GV C95: 4.86 log CFU/mL (72,836 CFU/mL) |
Aptima CV/TV Assay (250 Test Kit):
| Acceptance Criteria (LoD of 100-Test Kit) | Reported Performance (LoD Confirmation for 250-Test Kit) |
|---|---|
| C. albicans C95 (LoD): 4439 CFU/mL | C. albicans C95 (LoD): 4439 CFU/mL |
| C. parapsilosis C95 (LoD): 9416 CFU/mL | C. parapsilosis C95 (LoD): 9416 CFU/mL |
| C. tropicalis C95 (LoD): 811 CFU/mL | C. tropicalis C95 (LoD): 811 CFU/mL |
| C. dubliniensis C95 (LoD): 1176 CFU/mL | C. dubliniensis C95 (LoD): 1176 CFU/mL |
| C. glabrata LoD: 41 CFU/mL | C. glabrata LoD: 41 CFU/mL |
| T. vaginalis LoD: 0.0024 cells/mL | T. vaginalis LoD: 0.0024 cells/mL |
The conclusion states: "The performance study results demonstrate that the Aptima BV assay 250 Test Kit on the Panther system performs comparably to the predicate device that is currently marketed for the same intended use." and "The Aptima BV 100 Test Kit LoD was confirmed in the Aptima BV 250 Test Kit configuration." (Similar statements are made for the CV/TV assay).
2. Sample sized used for the test set and the data provenance (e.g. country of origin of the data, retrospective or prospective)
- Sample Size: For the LoD confirmation study for both assays, at least 20 replicates per concentration, per reagent lot, using three lots were tested. This totals to at least 60 replicates per strain (for BV) or per organism (for CV/TV).
- Data Provenance: The document does not specify the country of origin of the data or whether the study was retrospective or prospective. It describes analytical sensitivity studies using prepared dilutions of cell lysates/suspensions. For the CV/TV assay, the use of "Natural Vaginal Swab Matrix (NVSM)" and "Simulated Vaginal Swab Matrix (SVSM)" is mentioned for dilutions.
3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts (e.g. radiologist with 10 years of experience)
This information is not provided and is not applicable to the type of analytical study performed. The LoD confirmation uses known concentrations of target organisms, not clinical samples requiring expert interpretation for ground truth.
4. Adjudication method (e.g. 2+1, 3+1, none) for the test set
This information is not provided and is not applicable to an analytical LoD confirmation study.
5. 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
This information is not provided and is not applicable. The device is a diagnostic assay (in vitro nucleic acid amplification test), not an AI-assisted imaging device that would involve human readers.
6. If a standalone (i.e. algorithm only without human-in-the loop performance) was done
The device itself is a standalone diagnostic assay (an in vitro nucleic acid amplification test) run on an automated system (Panther system). Its performance (LoD) was confirmed without human interpretation of raw signals, as the Panther system software interprets the amplification signal emergence times to generate results.
7. The type of ground truth used (expert consensus, pathology, outcomes data, etc)
The ground truth used for this specific study (LoD confirmation) was known concentrations of purified target organisms/cell lysates. This is an analytical ground truth, not a clinical ground truth derived from expert consensus, pathology, or outcomes data.
8. The sample size for the training set
This information is not provided and is not applicable. This is not an AI/ML device that requires a training set in the conventional sense. The "training" for the assay involves internal optimization and validation during development, but the document does not detail these earlier stages.
9. How the ground truth for the training set was established
This information is not provided and is not applicable, as it's not an AI/ML device with a distinct training set and ground truth establishment methodology in the context of this submission.
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November 25, 2024
Image /page/0/Picture/1 description: The image shows 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 consists of a blue square with the letters "FDA" in white, followed by the words "U.S. FOOD & DRUG" in blue, and then the word "ADMINISTRATION" in a smaller font size below that.
Hologic, Inc. Gabriela Mccoole Regulatory Affairs Specialist 10210 Genetic Center Dr San Diego, California 92121
Re: K243345
Trade/Device Name: Aptima BV Assay: 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 Dated: October 23, 2024 Received: October 28, 2024
Dear Gabriela Mccoole:
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 (the 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 available 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.
Additional information about changes that may require a new premarket notification are provided in the FDA guidance documents entitled "Deciding When to Submit a 510(k) for a Change to an Existing Device"
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(https://www.fda.gov/media/99812/download) and "Deciding When to Submit a 510(k) for a Software Change to an Existing Device" (https://www.fda.gov/media/99785/download).
Your device is also subject to, among other requirements, the Quality System (QS) regulation (21 CFR Part 820), which includes, but is not limited to, 21 CFR 820.30. Design controls; 21 CFR 820.90. Nonconforming product; and 21 CFR 820.100, Corrective and preventive action. Please note that regardless of whether a change requires premarket review. the OS regulation requires device manufacturers to review and approve changes to device design and production (21 CFR 820.30 and 21 CFR 820.70) and document changes and approvals in the device master record (21 CFR 820.181).
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR Part 803) for devices or postmarketing safety reporting (21 CFR Part 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safetyreporting-combination-products); 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 Part 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR Parts 1000-1050.
All medical devices, including Class I and unclassified devices and combination product device constituent parts are required to be in compliance with the final Unique Device Identification System rule ("UDI Rue"). The UDI Rule requires, among other things, that a device bear a unique device identifier (UDI) on its label and package (21 CFR 801.20(a)) unless an exception or alternative applies (21 CFR 801.20(b)) and that the dates on the device label be formatted in accordance with 21 CFR 801.18. The UDI Rule (21 CFR 830.300(a) and 830.320(b)) also requires that certain information be submitted to the Global Unique Device Identification Database (GUDID) (21 CFR Part 830 Subpart E). For additional information on these requirements, please see the UDI System webpage at https://www.fda.gov/medical-devices/device-advicecomprehensive-regulatory-assistance/unique-device-identification-system-udi-system.
Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.
For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). 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 (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
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Sincerely,
Digitally signed by Bryan M. Bryan M. Grabias -S Date: 2024.11.25 Grabias -S 12:19:01 -05'00'
Bryan Grabias, Ph. D. Acting Branch Chief Bacterial Respiratory and Medical Countermeasures Branch Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics 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) K243345
Device Name Aptima BV Assay; Aptima CV/TV Assay
Indications for Use (Describe)
Aptima BV Assay
The Aptima BV Assay is an in vitro nucleic acid amplification test that utilizes real time Transcription-Mediated Amplification (TMA) technology for detection and quantitation of ribosomal RNA from bacterial vaginosis (BV), including Lactobacillus (L. gasseri, L. crispatus, and L. jenseni), Gardnerella vaginalis), and Atopobium vaginae (A. 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.
Aptima CV/TV Assay
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) technology to detect and qualitatively report results for the following organisms:
- · Candida species group (C. albicans, C. tropicalis, C. parapsilosis, C. dubliniensis)
- · Candida glabrata (C. glabrata)
- Trichomonas vaginalis (TV)
The assay differentiates between C. 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 TV, the assay targets ribosomal RNA (rRNA) and differentiates the results for C. glabrata and C spp. The assy is intended to aid in the diagnosis of vulvovaginal candidiasis and trichomoniasis on the automated Panther System using clinician-collected and patientcollected vaginal swab specimens from females with a clinical presentation consistent with vagintis.
Type of Use (Select one or both, as applicable)
X Prescription Use (Part 21 CFR 801 Subpart D)
| Over-The-Counter Use (21 CFR 801 Subpart C)
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Image /page/4/Picture/0 description: The image shows the word "HOLOGIC" in large, bold, blue letters. A small registered trademark symbol is located to the right of the letter "C". The font is sans-serif and the letters are evenly spaced.
510(k) SUMMARY Aptima BV Assay
I. SUBMITTER
Hologic, Inc. 10210 Genetic Center Drive San Diego, CA 92121
Contact Person: Gabriela McCoole, MS, RAC Regulatory Affairs Specialist Gabriela.McCoole@Hologic.com Phone: (619) 322-7535 Fax: (858) 410-5557
Date Prepared: October 25, 2024
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 PQA, NSU, and PMN Product Code:
III. PREDICATE DEVICE
The predicate device is the Aptima BV Assay (K190452; cleared May 23, 2019), which
is a 100 test kit configuration.
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DEVICE DESCRIPTION IV.
Like the Aptima BV assay 100 test kit, the Aptima BV assay 250 test kit 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.
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
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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.
Assay Components
The Aptima BV 250 test kit provides enough reagents to run 250 tests. Like the Aptima BV assay 100 test kit, the Aptima BV assay 250 test kit 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 (Aptima Assay Fluids Kit) required to run the assay, and one collection kit (Aptima Multitest Swab Specimen Collection Kit ) utilized for collection of specimens.
| 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 250 test kit | ||||
|---|---|---|---|---|
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Instrumentation
Like the Aptima BV assay 100 test kit, the 250 test kit configuration 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.
V. INDICATIONS FOR USE
The Aptima® BV Assay is an in vitro nucleic acid amplification test that utilizes real time Transcription-Mediated Amplification (TMA®) technology 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 (G. vaginalis), and Atropbium vaginae (A. 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.
COMPARISON OF TECHNOLOGICAL VI. CHARACTERISTCS WITH THE PREDICATE DEVICE
Table 2 displays a comparison of the Aptima BV Assay 250 Test Kit (subject device) with the Aptima BV Assay 250 Test Kit (predicate device).
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| Feature | Aptima BV Assay 100 Test Kit(Predicate Device, K190452) | Aptima BV Assay 250 Test Kit(Subject Device) Comparison to thePredicate | Predicate and SubjectDevice Comparison |
|---|---|---|---|
| Intended Use | The Aptima BV assay is an in vitro nucleic acidamplification test that utilizes real timetranscription-mediated amplification (TMA) fordetection and quantitation of ribosomal RNAfrom bacteria associated with bacterial vaginosis(BV), including Lactobacillus (L. gasseri, L.crispatus, and L. jensenii), Gardnerella vaginalis,and Atopobium vaginae. The assay reports aqualitative result for BV and does not reportresults for individual organisms. The assay isintended to aid in the diagnosis of BV on theautomated Panther system using clinician-collected and patient-collected vaginal swabspecimens from females with a clinicalpresentation consistent with vaginitis and/orvaginosis. | The Aptima® BV Assay is an in vitro nucleicacid amplification test that utilizes real timeTranscription-Mediated Amplification (TMA®)technology for detection and quantitation ofribosomal RNA from bacteria associated withbacterial vaginosis (BV), includingLactobacillus (L. gasseri, L. crispatus, and L.jensenii), Gardnerella vaginalis (G. vaginalis),and Atopobium vaginae (A. vaginae). The assayreports a qualitative result for BV and does notreport results for individual organisms. Theassay is intended to aid in the diagnosis of BVon the automated Panther® System usingclinician-collected and patient-collected vaginalswab specimens from females with a clinicalpresentation consistent with vaginitis and/orvaginosis. | Same; some editorialchanges were made to theintended use statement(added trademark symbolsand abbreviations forGardnerella vaginalis andAtopobium vaginae). |
| OrganismsDetected | Lactobacillus (L. gasseri, L. crispatus, and L.jensenii), Gardnerella vaginalis, and Atopobiumvaginae | Lactobacillus (L. gasseri, L. crispatus, and L.jensenii), Gardnerella vaginalis, and Atopobiumvaginae | Same |
| Test Quantity | 100 Tests | 250 Tests | Different; The subjectdevice quantity is 250 Tests. |
| Assay Controls | Incorporates an Internal Control in every test.Uses external positive and negative controls. | Incorporates an Internal Control in every test.Uses external positive and negative controls. | Same |
| Feature | Aptima BV Assay 100 Test Kit(Predicate Device, K190452) | Aptima BV Assay 250 Test Kit(Subject Device) Comparison to thePredicate | Predicate and SubjectDevice Comparison |
| PatientPopulation | Women with a clinical presentation ofvaginitis/vaginosis | Women with a clinical presentation ofvaginitis/vaginosis | Same |
| Specimen Types | Vaginal swabs in patients who are symptomaticfor vaginitis/vaginosis | Vaginal swabs in patients who are symptomaticfor vaginitis/vaginosis | Same |
| Analyte | ribosomal RNA | ribosomal RNA | Same |
| Assay Calibrators | Positive Calibrator | Positive Calibrator | Same |
| TechnologyPrinciple ofOperation | Real-time Transcription Mediated Amplification(TMA) | Real-time Transcription Mediated Amplification(TMA) | Same |
| User Complexity | High | High | Same |
| Platform | Panther System | Panther System | Same |
| PlatformSoftware | Panther System Software | Panther System Software | Same |
| Assay Software | Panther Aptima BV 100 Test Kit AssaySoftware | Panther Aptima BV 250 Test Kit AssaySoftware. | Different. The subjectdevice uses the PantherAptima BV 250 Test KitAssay Software. |
Table 2 Comparison of Predicate and Subject Device
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PERFORMANCE DATA VII.
The Aptima BV assay 100 Test Kit and the 250 Test Kit (subject device) share the same intended use, operating principle, formulation, and accessories. Based on these similarities, a risk analysis determined the analytical studies performed for the 100 Test Kit also support the Aptima BV 250 Test Kit configuration performance. A Limit of Detection (LoD) Confirmation study was performed to verify that the Aptima BV 250 Test Kit configuration meets the established Aptima BV 100 Test Kit LoD. The Aptima BV 100 Test Kit LoD was confirmed in the Aptima BV 250 Test Kit configuration, and thus all other previously validated analytical and clinical performance testing performed with the Aptima BV 100 Test Kit configuration is valid and appropriate to show performance for the Aptima BV 250 Test Kit configuration.
Analytical Sensitivity
To confirm the Limit of Detection (LoD) for L. crispatus (LC), L. gasseri (LG), L. jensenii (LJ) and the BV Positivity Limit (C95) for A.vaginae (AV) and G. vaginalis (GV), dilutions of cell lysates were prepared in SVSM. Dilutions were then tested with at least 20 replicates per concentration, per reagent lot, using three lots over the course of three days (7 replicates per day) for a total of at least 60 replicates per strain. The results verified that the LoD for each strain is as follows: LC is 143 CFU/mL, LG is 2,207 CFU/mL, and LJ is 10 CFU/mL. The BV Positivity Limit (C95) for AV and GV were verified to be 5.10 log CFU/mL (128,397 CFU/mL), and 4.86 log CFU/mL (72,836 CFU/mL) respectively.
CONCLUSIONS VIII.
The performance study results demonstrate that the Aptima BV assay 250 Test Kit 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 250 Test Kit on the Panther system will perform as intended.
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Image /page/11/Picture/0 description: The image shows the word "HOLOGIC" in a bold, sans-serif font. The letters are a dark blue color. There is a registered trademark symbol to the upper right of the "C".
510(k) SUMMARY Aptima CV/TV Assay
I. SUBMITTER
Hologic, Inc. 10210 Genetic Center Drive San Diego, CA 92121
Contact Person: Gabriela McCoole, MS, RAC Regulatory Affairs Specialist Gabriela.McCoole@Hologic.com Phone: (619) 322-7535 Fax: (858) 410-5557
Date Prepared: October 25, 2024
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 Aptima CV/TV Assay (K190472; cleared February 26, 2019),
which is a 100 test kit configuration.
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DEVICE DESCRIPTION IV.
Like the Aptima CV/TV assay 100 test kit, the Aptima CV/TV assay 250 test kit is an in vitro nucleic acid amplification test for the detection and quantitation of RNA from microorganisms associated with vaginitis, trichomoniasis, and vulvovaginitis, in women with a clinical presentation consistent with vaginitis, trichomoniasis, and 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.
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.
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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 Cspp, 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
Assay Components
The Aptima CV/TV 250 test kit provides enough reagents to run 250 tests. Like the Aptima CV/TV assay 100 test kit, the Aptima CV/TV assay 250 test kit 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 (Aptima Assay Fluids Kit) required to run the assay, and one collection kit (Aptima Multitest Swab Specimen Collection Kit ) utilized for collection of specimens.
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| 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 CV/TV Assay 250 test kit
Instrumentation
Like the Aptima CV/TV assay 100 test kit, the 250 test kit configuration 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.
INDICATIONS FOR USE V.
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®) technology to detect and qualitatively report results for the following organisms:
- Candida species group (C. albicans, C. tropicalis, C. parapsilosis, C. dubliniensis)
- Candida glabrata (C. glabrata) ●
- Trichomonas vaginalis (TV)
The assay differentiates between C. 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 TV, the assay targets ribosomal RNA (rRNA) and
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differentiates the result from results for C. 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.
COMPARISON OF TECHNOLOGICAL VI. CHARACTERISTCS WITH THE PREDICATE DEVICE
Table 2 displays a comparison of the Aptima CV/TV Assay 250 Test Kit (subject device) with the Aptima CV/TV Assay 250 Test Kit (predicate device).
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| Item | Aptima CV/TV Assay 100 Test Kit(Predicate Device, K190472) | Aptima CV/TV Assay 250 Test Kit(Subject Device) | Predicate and SubjectDevice Comparison |
|---|---|---|---|
| Intended Use | The Aptima CV/TV assay is an in vitro nucleic acidamplification test for the detection of RNA frommicroorganisms associated with vaginitis,trichomoniasis, and vulvovaginitis. The assayutilizes real time transcription-mediatedamplification (TMA) to detect and qualitativelyreport results for the following organisms:• Candida species group (C. albicans, C. tropicalis,C. parapsilosis, C. dubliniensis)• Candida glabrata• Trichomonas vaginalisThe assay differentiates between Candida glabrataand the Candida species group (C spp) by targetingthe RNA component of RNAse P ribonucleoprotein;the assay does not differentiate among C spp. ForTrichomonas vaginalis, the assay targets ribosomalRNA (rRNA) and differentiates the result fromresults for Candida glabrata and C spp. The assay isintended to aid in the diagnosis of vulvovaginalcandidiasis and trichomoniasis on the automatedPanther system using clinician-collected and patient-collected vaginal swab specimens from women witha clinical presentation consistent with vaginitis,trichomoniasis, and/or vulvovaginitis. | The Aptima® CV/TV Assay is an in vitro nucleicacid amplification test for the detection of RNAfrom microorganisms associated withvulvovaginal candidiasis and trichomoniasis. Theassay utilizes real time Transcription-MediatedAmplification (TMA®) technology to detect andqualitatively report results for the followingorganisms:• Candida species group (C. albicans, C.tropicalis, C. parapsilosis, C.dubliniensis)• Candida glabrata (C. glabrata)• Trichomonas vaginalis (TV)The assay differentiates between C. glabrata andthe Candida species group (C spp) by targetingthe RNA component of RNAse Pribonucleoprotein; the assay does notdifferentiate among C spp. For TV, the assaytargets ribosomal RNA (rRNA) and differentiatesthe result from results for C. glabrata and C spp.The assay is intended to aid in the diagnosis ofvulvovaginal candidiasis and trichomoniasis onthe automated Panther® System using clinician-collected and patient-collected vaginal swabspecimens from females with a clinicalpresentation consistent with vaginitis orvulvovaginitis. | Same. Added trademarksand Candida glabrata andTrichomonas vaginalisabbreviations. |
| Item | Aptima CV/TV Assay 100 Test Kit(Predicate Device, K190472) | Aptima CV/TV Assay 250 Test Kit(Subject Device) | Predicate and SubjectDevice Comparison |
| OrganismsDetected | Candida species (C. albicans, C. tropicalis, C.parapsilosis, C. dubliniensis); Candida glabrata;Trichomonas vaginalis | Candida species (C. albicans, C. tropicalis, C.parapsilosis, C. dubliniensis); Candida glabrata;Trichomonas vaginalis | Same |
| Test Quantity | 100 Tests | 250 Tests | Different.The subject devicequantity is 250 Tests. |
| AssayControls | Incorporates an Internal Control in every test. Usesexternal positive and negative controls. | Incorporates an Internal Control in every test. Usesexternal positive and negative controls. | Same |
| PatientPopulation | Women with a clinical presentation of vaginitis,trichomoniasis, or vulvovaginitis | Women with a clinical presentation of vaginitis,trichomoniasis, or vulvovaginitis | Same |
| SpecimenTypes | Vaginal swabs in patients who are symptomatic forvaginitis, trichomoniasis, or vulvovaginitis | Vaginal swabs in patients who are symptomatic forvaginitis, trichomoniasis, or vulvovaginitis | Same |
| Analyte | ribosomal RNA | ribosomal RNA | Same |
| AssayCalibrators | Positive Calibrator | Positive Calibrator | Same |
| TechnologyPrinciple ofOperation | Real-time Transcription Mediated Amplification(TMA) | Real-time Transcription Mediated Amplification(TMA) | Same |
| UserComplexity | High | High | Same |
| Item | Aptima CV/TV Assay 100 Test Kit(Predicate Device, K190472) | Aptima CV/TV Assay 250 Test Kit(Subject Device) | Predicate and SubjectDevice Comparison |
| Platform | Panther System | Panther System | Same |
| PlatformSoftware | Panther System Software | Panther System Software | Same |
| AssaySoftware | Panther Aptima CV/TV 100 Test Kit AssaySoftware | Panther Aptima CV/TV 250 Test Kit AssaySoftware | Different.The subject device usesPanther Aptima CV/TV250 Test Kit AssaySoftware. |
Table 2 Comparison of Predicate and Subject Device
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PERFORMANCE DATA VII.
The Aptima CV/TV assay 100 Test Kit and the 250 Test Kit (subject device) share the same intended use, operating principle, formulation, and accessories. Based on these similarities, a risk analysis determined the analytical studies performed for the 100 Test Kit also support the Aptima CV/TV 250 Test Kit configuration performance. A Limit of Detection (LoD) Confirmation study was performed to verify that the Aptima CV/TV 250 Test Kit configuration meets the established Aptima CV/TV 100 Test Kit LoD. The Aptima CV/TV 100 Test Kit LoD was confirmed in the Aptima CV/TV 250 Test Kit configuration, and thus all other previously validated analytical and clinical performance testing performed with the Aptima CV/TV 100 Test Kit configuration is valid and appropriate to show performance for the Aptima CV/TV 250 Test Kit configuration.
Analytical Sensitivity
To confirm the Limit of Detection (LoD) or C95 of C. albicans, C. glabrata, and T. vaginalis, cell lysates were diluted into Natural Vaginal Swab Matrix (NVSM). Cell lysates of C. tropicalis and C. dubliniensis and suspension of C. parapsilosis were diluted into Simulated Vaginal Swab Matrix (SVSM). Dilutions were then tested with at least 20 replicates per concentration, per reagent lot, using three lots for a total of at least 60 replicates per strain.
The results verified that the LoD for each strain is as follows: The C95 (LoD) for C. albicans is 4439 CFU/mL, for C. parapsilosis is 9416 CFU/mL, for C. tropicalis is 811 CFU/mL, for C. dubliniensis is 1176 CFU/mL. The LoD for C. glabrata is 41 CFU/mL, and for T. vaginalis is 0.0024 cells/mL.
VIII. CONCLUSIONS
The performance study results demonstrate that the Aptima CV/TV assay 250 Test Kit 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
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demonstrate that the Aptima CV/TV assay 250 Test Kit 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.