(262 days)
No
The description details a standard nucleic acid amplification test (NAAT) with real-time detection based on fluorescence. The data analysis involves comparing fluorescence curves to fixed cut-off times, which is a deterministic algorithm, not AI/ML. There is no mention of AI, ML, or related concepts in the document.
No
This device is an in vitro diagnostic test for the detection of HSV, intended as an aid in diagnosis, not for treating or preventing disease.
Yes
The "Intended Use / Indications for Use" section explicitly states, "The Aptima Herpes Simplex Viruses 1 & 2 assay... is an in vitro diagnostic nucleic acid amplification test (NAAT)... The assay is intended for use as an aid in the diagnosis of HSV-1 and/or HSV-2 infections in symptomatic male and female patients."
No
The device is an in vitro diagnostic nucleic acid amplification test (NAAT) that utilizes reagents and the Panther® system hardware for sample processing, amplification, and detection. While software is involved in the analysis of results, the core functionality relies on physical components and chemical reactions.
Yes, this device is an IVD (In Vitro Diagnostic).
The text explicitly states in the "Intended Use / Indications for Use" section: "The Aptima Herpes Simplex Viruses 1 & 2 assay (Aptima HSV 1 & 2 assay) is an in vitro diagnostic nucleic acid amplification test (NAAT)..."
N/A
Intended Use / Indications for Use
The Aptima Herpes Simplex Viruses 1 & 2 assay (Aptima HSV 1 & 2 assay) is an in vitro diagnostic nucleic acid amplification test (NAAT), using real time transcription-mediated amplification (TMA), for the qualitative detection and differentiation of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) messenger RNA (mRNA) in clinician-collected swab specimens from anogenital skin lesions. The assay is intended for use with swab specimens placed in Aptima specimen transport medium (STM) or in viral transport media (VTM) that is immediately diluted into STM.
The Aptima HSV 1 & 2 assay is intended for use as an aid in the diagnosis of HSV-1 and/or HSV-2 infections in symptomatic male and female patients. The Aptima HSV 1 & 2 assay is indicated for use on the Panther® system.
Warning
The Aptima HSV 1 & 2 assay is not FDA-cleared for use with cerebrospinal fluid (CSF) or for prenatal screening.
Product codes
OQO
Device Description
The Aptima Herpes Simplex Virus 1 & 2 assay (Aptima HSV assay) is a nucleic acid amplification test (NAAT) developed for use on the fully automated Panther system that utilizes target capture, transcription mediated amplification (TMA), and real-time detection of HSV-1, HSV-2, and an internal control (IC). The Aptima HSV assay amplifies and detects mRNAs for HSV-1 and HSV-2. These RNAs are expressed from the viral genome during the infection cycle, and are packaged inside HSV-1 and HSV-2 viral particles prior to virus release from infected cells. The Aptima HSV assay therefore detects virus-infected cells and the mature virus particles themselves.
The Aptima HSV assay involves three main steps, which all take place in a single tube on the Panther® system: target capture, target amplification by TMA, and detection of the amplification products (amplicon) by the fluorescent labeled probes (torches). The assay incorporates an IC in every test to monitor targeted nucleic acid capture, amplification and detection.
When the Aptima HSV assay is performed, the targeted viral mRNA and IC are isolated using magnetic microparticles and target-specific capture oligomers, in a process called target capture. The capture oligomers contain sequences complementary to specific regions of the targeted RNA (HSV mRNA or IC) as well as a string of deoxyadenosine residues. During the hybridization step, the sequence-specific regions of the capture oligomers bind to specific regions of the RNA target molecules. The microparticles, including the captured RNA target molecules bound to them, are pulled to the side of the reaction tube using magnets and the supernatant is aspirated. The particles are washed to remove residual specimen matrix that may contain amplification inhibitors. After target capture steps are completed, the specimens are ready for amplification.
Target amplification occurs via TMA, which is a transcription-based nucleic acid amplification method that utilizes two enzymes, MMLV (Moloney murine leukemia virus) reverse transcriptase and T7 RNA polymerase. The reverse transcriptase is used to generate a DNA copy (containing a promoter sequence for T7 RNA polymerase) of the target sequence. 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 as it is designed to be in close proximity when not hybridized to the amplicon. When the torch binds to the amplicon, the quencher is moved farther away from the fluorophore and it will emit a signal at a specific wavelength when excited by a light source. More torch hybridizes when more amplicon is present. The increase in fluorescent signal from progressive amplification is detected by fluorometers within the Panther system. The Panther system can detect and discriminate between the three fluorescent signals corresponding to HSV-1, HSV-2 and IC amplification products. The fluorescence (measured in relative fluorescence units [RFU]) is monitored over time to produce a real-time fluorescence emergence curve for each reporter dye. The Panther system software compares the fluorescence emergence curves to fixed cut off times to report results (TTime) for HSV-1, HSV-2 and IC.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
anogenital skin lesions
Indicated Patient Age Range
Not Found
Intended User / Care Setting
in clinical laboratories by trained laboratory professionals
Description of the training set, sample size, data source, and annotation protocol
Not Found
Description of the test set, sample size, data source, and annotation protocol
Not Found
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Analytical Sensitivity and Limit of Detection (LoD)
The LoD in clinical specimen matrix was determined by testing dilution panels for each of two strains of HSV-1 virus (Macintyre and HF strains) and two strains of HSV-2 viral (MS and G) in both Aptima Specimen Transport Media (STM) and Viral Transport Media (VTM) negative clinical pools in the Aptima HSV assay. LoD for the assay was based on results from the reagent lot with the highest concentration for the 95% predicted detection limit. The probit-predicted 95% LoD values for HSV-1 ranged from 60.6 TCID50/mL (STM) to 186.9 TCID50/mL (VTM). For HSV-2, the LoD ranged from 18.2 TCID50/mL (STM) to 128.8 TCID50/mL (VTM).
Limit of Detection Verification
LoD of the Aptima HSV 1 & 2 assay was verified using two clinical isolates of HSV-1 and two clinical isolates of HSV-2. Each isolate was tested with the Aptima HSV 1 & 2 assay using 60 replicates each at 1X LoD, 3X LoD, and 10X LoD. Testing was completed in both STM and VTM matrix for all four clinical isolates and was conducted using 3 lots of reagents. All replicates for all clinical isolates at all three concentrations tested were detected by the Aptima HSV 1 & 2 assay.
Limit of Detection in Neat STM
The LoD in neat STM was determined by testing panels of HSV-1 and HSV-2 in vitro RNA transcripts (IVT) and viral particles. Results show a predicted 95% LoD of 68.3 copies/mL for HSV-1 IVT and 49.4 copies/mL for HSV-2 IVT. For viral particles, the predicted 95% LoD was 28.9 TCID50/mL for HSV-1 MacIntyre virus and 0.54 TCID50/mL for HSV-2 MS virus.
Interfering Substances
Potentially interfering substances were tested. No effect on assay performance was observed in the presence of various exogenous and endogenous substances at specified concentrations.
Co-Infection
A co-infection study was evaluated to assess the ability of the Aptima HSV 1 & 2 assay to detect HSV 1 and HSV 2 analytes when both are present in one specimen. Low and high titer concentrations of HSV-1 viral particles in STM were tested in combination with low, moderate, and high concentrations of HSV-2 virus. All testing resulted in 100% detection for both HSV-1 and HSV-2.
Cross-Reactivity
A study was performed to evaluate the cross-reactivity and interference with the Aptima HSV 1 & 2 assay in the presence of non-targeted microorganisms (48 microorganisms). No evidence of cross-reactivity or microbial interference was observed for any of the 48 test microorganisms, except for Streptococcus pneumoniae at 1x10^6 CFU/mL where cross-reactivity was observed.
Specimen Inhibition in Clinical Samples
An evaluation was conducted of the whole system failure rate by testing low positive specimens. Of 201 STM specimens tested, the false negative rate was 0% (0/201) for HSV-1 and 0.5% (1/201) for HSV-2. Of 202 VTM specimens tested, the false negative rate was 0% (0/202) for HSV-1 and 0% (0/202) for HSV-2.
Reproducibility and Within Laboratory Precision
One HSV negative panel and six HSV positive panels were tested using STM. Three replicates of each panel were tested by three operators in two runs each in three reagent lots over 13 days. Results showed acceptable precision and reproducibility.
Clinical Performance Study
A prospective, multicenter clinical study was conducted.
- Study Type: Clinical performance study with a composite reference method.
- Sample Size: 544 evaluable subjects (195 males and 349 females) with active anogenital lesions.
- Data Source: Subjects enrolled from 17 US clinical sites, including family planning, dermatology, pediatrics/adolescent, sexually transmitted infection, private practice, public health clinics, hospitals, universities, and clinical research sites.
- Annotation Protocol: Two swab specimens collected per subject (one in VTM and one in STM). The performance of the Aptima HSV-1 & 2 assay was evaluated relative to a composite reference, which used results from ELVIS HSV ID and D3 Typing Test system viral culture and a validated bidirectional PCR/sequencing procedure. A third FDA-cleared assay was used to determine the final composite reference interpretation when the ELVIS D3 culture and PCR/sequencing results did not agree or when PCR/sequencing detected both HSV-1 and HSV-2.
- Key Results:
- For HSV-1:
- VTM: Sensitivity 93.4% (95% CI: 85.5-97.2), Specificity 99.8% (95% CI: 98.8-99.9), PPV 98.6% (95% CI: 93.0-100), NPV 98.9% (95% CI: 97.6-99.6).
- STM: Sensitivity 94.7% (95% CI: 87.1-97.9), Specificity 99.6% (95% CI: 98.4-99.9), PPV 97.3% (95% CI: 91.1-99.6), NPV 99.1% (95% CI: 97.9-99.8).
- For HSV-2:
- VTM: Sensitivity 96.9% (95% CI: 94.0-98.4), Specificity 97.5% (95% CI: 94.9-98.8), PPV 97.3% (95% CI: 94.7-98.8), NPV 97.1% (95% CI: 94.6-98.7).
- STM: Sensitivity 98.4% (95% CI: 96.1-99.4), Specificity 92.8% (95% CI: 89.1-95.3), PPV 92.7% (95% CI: 89.4-95.3), NPV 98.5% (95% CI: 96.3-99.6).
- For HSV-1:
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
HSV-1 Performance (Combined Gender, All Ages, Clinical Performance Study):
- VTM Specimen Type:
- Sensitivity: 93.4% (95% CI: 85.5-97.2)
- Specificity: 99.8% (95% CI: 98.8-99.9)
- PPV: 98.6% (95% CI: 93.0-100)
- NPV: 98.9% (95% CI: 97.6-99.6)
- STM Specimen Type:
- Sensitivity: 94.7% (95% CI: 87.1-97.9)
- Specificity: 99.6% (95% CI: 98.4-99.9)
- PPV: 97.3% (95% CI: 91.1-99.6)
- NPV: 99.1% (95% CI: 97.9-99.8)
HSV-2 Performance (Combined Gender, All Ages, Clinical Performance Study):
- VTM Specimen Type:
- Sensitivity: 96.9% (95% CI: 94.0-98.4)
- Specificity: 97.5% (95% CI: 94.9-98.8)
- PPV: 97.3% (95% CI: 94.7-98.8)
- NPV: 97.1% (95% CI: 94.6-98.7)
- STM Specimen Type:
- Sensitivity: 98.4% (95% CI: 96.1-99.4)
- Specificity: 92.8% (95% CI: 89.1-95.3)
- PPV: 92.7% (95% CI: 89.4-95.3)
- NPV: 98.5% (95% CI: 96.3-99.6)
Prevalence (Clinical Study):
- VTM: HSV-1: 13.6% (72/528), HSV-2: 47.8% (255/533)
- STM: HSV-1: 13.7% (73/531), HSV-2: 51.0% (273/535)
Predicate Device(s)
Reference Device(s)
K003395, K032554, K063664, K063451, K043224, K122062, K111409
Predetermined Change Control Plan (PCCP) - All Relevant Information
Not Found
§ 866.3305 Herpes simplex virus serological assays.
(a)
Identification. Herpes simplex virus serological assays are devices that consist of antigens and antisera used in various serological tests to identify antibodies to herpes simplex virus in serum. Additionally, some of the assays consist of herpes simplex virus antisera conjugated with a fluorescent dye (immunofluorescent assays) used to identify herpes simplex virus directly from clinical specimens or tissue culture isolates derived from clinical specimens. The identification aids in the diagnosis of diseases caused by herpes simplex viruses and provides epidemiological information on these diseases. Herpes simplex viral infections range from common and mild lesions of the skin and mucous membranes to a severe form of encephalitis (inflammation of the brain). Neonatal herpes virus infections range from a mild infection to a severe generalized disease with a fatal outcome.(b)
Classification. Class II (special controls). The device is classified as class II (special controls). The special control for the device is FDA's revised guidance document entitled “Class II Special Controls Guidance Document: Herpes Simplex Virus Types 1 and 2 Serological Assays.” For availability of the guidance revised document, see § 866.1(e).
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Public Health Service
Image /page/0/Picture/2 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features a stylized depiction of an eagle or bird-like figure, composed of three curved lines that suggest the head and body. The logo is encircled by the text "DEPARTMENT OF HEALTH & HUMAN SERVICES • USA" in a circular arrangement.
Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
June 15, 2017
Hologic, Inc. Ron Domingo Regulatory Affairs Manager 10210 Genetic Center Dr. San Diego, California 92121
Re: K162673
Trade/Device Name: Aptima Herpes Simplex Viruses 1 & 2 Assay Regulation Number: 21 CFR 866.3305 Regulation Name: Herpes simplex virus serological assays Regulatory Class: Class II Product Code: OOO Dated: May 16, 2017 Received: May 17, 2017
Dear Mr. Domingo:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration. Isting of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements
1
as set forth in the quality systems (OS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
If vou desire specific advice for your device on our labeling regulation (21 CFR Part 801 and Part 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to
http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.
You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.
Sincerely,
Stephen J. Lovell –S for
Uwe Scherf, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
2
Indications for Use
510(k) Number (if known) K162673
Device Name Aptima Herpes Simplex Viruses 1 & 2 Assay
Indications for Use (Describe)
The Aptima Herpes Simplex Viruses 1 & 2 assay (Aptima HSV 1 & 2 assay) is an in vitro diagnostic nucleic acid amplification test (NAAT), using real time transcription-mediated amplification (TMA), for the qualitative detection and differentiation of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) messenger RNA (mRNA) in clinician-collected swab specimens from anogenital skin lesions. The assay is intended for use with swab specimens placed in Aptima specimen transport medium (STM) or in viral transport media (VTM) that is immediately diluted into STM.
The Aptima HSV 1 & 2 assay is intended for use as an aid in the diagnosis of HSV-1 and/or HSV-2 infections in symptomatic male and female patients. The Aptima HSV 1 & 2 assay is indicated for use on the Panther® system.
Warning
The Aptima HSV 1 & 2 assay is not FDA-cleared for use with cerebrospinal fluid (CSF) or for prenatal screening.
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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3
510(k) SUMMARY Aptima Herpes Simplex Viruses 1 & 2 Assay
I. SUBMITTER
Hologic, Inc. 10210 Genetic Center Drive San Diego, CA 92121
| Contact Person: | Ron Domingo, MS, RAC
Regulatory Affairs Manager |
|-----------------|----------------------------------------------------|
| | Ron.Domingo@Hologic.com |
| | Phone: (858) 410-8167 |
| | Fax: (858) 410-5557 |
Date Prepared: June 14, 2017
II. DEVICE
| Proprietary Name of Device: | Aptima Herpes Simplex Viruses 1 & 2 Assay | |
|---|---|---|
| Common or Usual Name: | Nucleic acid amplification test for Herpes Simplex viruses | |
| Classification Name: | Herpes simplex virus serological assays | |
| Regulation Number: | 21 CFR 866.3305 | |
| Regulatory Class: | Class II | |
| Product Code: | OQO |
III. PREDICATE DEVICE
The predicate device is the BD ProbeTec Herpes Simplex Viruses (HSV 1 & 2) Q* Amplified DNA Assays (K103798; cleared March 18, 2011; BD Diagnostics Systems, Sparks, MD).
IV. DEVICE DESCRIPTION
The Aptima Herpes Simplex Virus 1 & 2 assay (Aptima HSV assay) is a nucleic acid amplification test (NAAT) developed for use on the fully automated Panther system that utilizes target capture, transcription mediated amplification (TMA), and real-time detection of HSV-1, HSV-2, and an internal control (IC). The Aptima HSV assay amplifies and detects mRNAs for
4
HSV-1 and HSV-2. These RNAs are expressed from the viral genome during the infection cycle, and are packaged inside HSV-1 and HSV-2 viral particles prior to virus release from infected cells. The Aptima HSV assay therefore detects virus-infected cells and the mature virus particles themselves.
The Aptima HSV assay involves three main steps, which all take place in a single tube on the Panther® system: target capture, target amplification by TMA, and detection of the amplification products (amplicon) by the fluorescent labeled probes (torches). The assay incorporates an IC in every test to monitor targeted nucleic acid capture, amplification and detection.
When the Aptima HSV assay is performed, the targeted viral mRNA and IC are isolated using magnetic microparticles and target-specific capture oligomers, in a process called target capture. The capture oligomers contain sequences complementary to specific regions of the targeted RNA (HSV mRNA or IC) as well as a string of deoxyadenosine residues. During the hybridization step, the sequence-specific regions of the capture oligomers bind to specific regions of the RNA target molecules. The microparticles, including the captured RNA target molecules bound to them, are pulled to the side of the reaction tube using magnets and the supernatant is aspirated. The particles are washed to remove residual specimen matrix that may contain amplification inhibitors. After target capture steps are completed, the specimens are ready for amplification.
Target amplification occurs via TMA, which is a transcription-based nucleic acid amplification method that utilizes two enzymes, MMLV (Moloney murine leukemia virus) reverse transcriptase and T7 RNA polymerase. The reverse transcriptase is used to generate a DNA copy (containing a promoter sequence for T7 RNA polymerase) of the target sequence. 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 as it is designed to be in close proximity when not hybridized to the amplicon. When the torch binds to the amplicon, the quencher is moved farther away from the fluorophore and it will emit a signal at a specific wavelength when excited by a light source. More torch hybridizes when more
5
amplicon is present. The increase in fluorescent signal from progressive amplification is detected by fluorometers within the Panther system. The Panther system can detect and discriminate between the three fluorescent signals corresponding to HSV-1, HSV-2 and IC amplification products. The fluorescence (measured in relative fluorescence units [RFU]) is monitored over time to produce a real-time fluorescence emergence curve for each reporter dye. The Panther system software compares the fluorescence emergence curves to fixed cut off times to report results (TTime) for HSV-1, HSV-2 and IC.
Kit Components
The Aptima HSV assay is provided as a 100 test kit. The following 3 kits are required to perform the Aptima HSV assay on the Panther system:
- Aptima Herpes Simplex Viruses 1 & 2 Assay Master Kit: Consists of 2 boxes containing eight reagents packaged by storage conditions (1 refrigerated and 1 room temperature box).
- • Aptima Herpes Simplex Viruses 1 & 2 Controls Kit: Consists of 1 box containing a Positive and Negative Control.
- Aptima Assay Fluids Kit: This ancillary kit is required to run the assay but is procured . separately. The Aptima Assay Fluids Kit consists of 1 box containing 3 components. The ancillary kit previously received market clearance for use in other commercialized Aptima assays (Ref: K003395).
The Aptima HSV assay utilizes two collection and handling kits, which are sold independently of the assay. The collection kits described below have previously received market clearance for use in other commercialized Aptima assays:
- Aptima Multitest Swab Specimen Collection Kit (Ref: K032554) .
- . Aptima Specimen Transfer Kit (Ref: K063664, K063451, K043224)
Instrumentation
The Aptima HSV assay has been designed for and validated on the Panther system. The Panther system is an integrated hardware and software system that fully automates all steps of nucleic
6
acid testing necessary for diagnostic assays. The instrument provides automation of all assay steps including sample processing, amplification of nucleic acid, detection and quantitation, amplicon inactivation, and data reduction. The "Panther Instrument" refers to the analyzer and associated hardware combined with the software. The "Panther system" refers to the Panther Instrument operated in conjunction with the Aptima HSV Assay. The assay software in the computer workstation directs the analyzer modules to perform each sequential assay step.
The Panther system was previously FDA cleared for use with other Class II Aptima Assays (Ref: K122062 and K111409).
V. INDICATIONS FOR USE
Intended Use
The Aptima Herpes Simplex Viruses 1 & 2 assay (Aptima HSV 1 & 2 assay) is an in vitro diagnostic nucleic acid amplification test (NAAT), using real time transcription-mediated amplification (TMA), for the qualitative detection and differentiation of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) messenger RNA (mRNA) in clinician-collected swab specimens from anogenital skin lesions. The assay is intended for use with swab specimens placed in Aptima specimen transport medium (STM) or in viral transport media (VTM) that is immediately diluted into STM.
The Aptima HSV 1 & 2 assay is intended for use as an aid in the diagnosis of HSV-1 and/or HSV-2 infections in symptomatic male and female patients. The Aptima HSV 1 & 2 assay is indicated for use on the Panther® system.
Warning
The Aptima HSV 1 & 2 assay is not FDA-cleared for use with cerebrospinal fluid (CSF) or for prenatal screening.
7
VI. COMPARISON OF TECHNOLOGICAL CHARACTERISTICS WITH THE PREDICATE DEVICE
A comparison of the Aptima Herpes HSV assay to the predicate BD ProbeTec Herpes Simplex Viruses (HSV 1 & 2) Q* Amplified DNA Assays (K103798) is summarized in Table 1 (similarities) and Table 2 (differences).
| Item | Aptima HSV Assay
(Subject Device)
K162673 | BD ProbeTec Assay
(Predicate Device)
K103798 |
|------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended Use | The Aptima Herpes Simplex
Viruses 1 & 2 assay (Aptima HSV
1 & 2 assay) is an in vitro
diagnostic nucleic acid
amplification test (NAAT), using
real time transcription-mediated
amplification (TMA), for the
qualitative detection and
differentiation of herpes simplex
virus type 1 (HSV-1) and type 2
(HSV-2) messenger RNA (mRNA)
in clinician-collected swab specimens
from anogenital skin lesions. The
assay is intended for use with swab
specimens placed in Aptima
specimen transport medium (STM)
or in viral transport media (VTM)
that is immediately diluted into
STM.
The Aptima HSV 1 & 2 assay is
intended for use as an aid in the
diagnosis of HSV-1 and/or HSV-2
infections in symptomatic male and
female patients. The Aptima HSV 1
& 2 assay is indicated for use on
the Panther® system.
Warning
The Aptima HSV 1 & 2 assay is
not FDA-cleared for use with
cerebrospinal fluid (CSF) or for | The BD ProbeTec Herpes
Simplex Viruses (HSV 1 & 2)
QX Amplified DNA Assays,
when tested with the BD Viper
System in Extracted Mode, use
Strand Displacement
Amplification technology for
the direct, qualitative detection
and differentiation of Herpes
Simplex virus type 1 (HSV1)
and Herpes Simplex virus type
2 (HSV2) DNA in clinician-
collected external anogenital
lesion specimens. The assays
are indicated for use with
symptomatic individuals to aid
in the diagnosis of anogenital
HSV1 and HSV2 infections.
Warning: The BD ProbeTec
Herpes Simplex Viruses (HSV
1 & 2) QX Amplified DNA
Assays (HSV QX Assays) are
not FDA cleared for use with
cerebrospinal fluid (CSF). The
assays are not intended to be
used for prenatal screening or
for individuals under the age of
17 years. |
| Item | Aptima HSV Assay
(Subject Device)
K162673 | BD ProbeTec Assay
(Predicate Device)
K103798 |
| Qualitative
/Quantitative Assay | Qualitative | Qualitative |
| Controls | Positive and negative controls.
Provided as part of the Master Kit. | Per PI, positive and negative
controls. Provided as part of a
Master Kit. |
| Patient Population | Symptomatic male and female
patients | Symptomatic male and female
patients |
| Specimen Types | Clinician-collected swab specimens
from skin lesions in the anogenital
region. | Clinician-collected external
anogenital lesion specimens. |
Table 1: Comparison of Similarities Between Aptima HSV Assay and Predicate Device
8
| Table 2: Comparison of Differences Between Aptima HSV Assav and Predicate Device | ||||
|---|---|---|---|---|
| Item | Aptima HSV Assay
(Subject Device) | BD ProbeTec Assay
(Predicate Device) |
|------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Target | mRNA | DNA |
| Function | Qualitative detection and
differentiation of mRNA from
herpes simplex viruses 1 & 2 | Qualitative detection and
differentiation of DNA from
herpes simplex viruses 1 & 2 |
| Technology | The subject assay is comprised of a
nucleic acid amplification test
(NAAT) for use on the automated
Panther system, that utilizes target
capture to extract viral mRNA from
patient samples, and real-time
transcription mediated
amplification (TMA) and detection
of HSV-1, and HSV-2 mRNAs and
an internal control RNA. | The predicate assay is
comprised of an amplification
test for use on the automated
BD Viper System that utilizes
non-specific extraction of DNA
from clinical specimens,
followed by binding of DNA to
magnetic particles, washing of
the bound nucleic acid and
elution in an amplification-
compatible buffer. When
present, HSV1 and/or 2 DNA is
detected by Strand
Displacement Amplification
(SDA) of type-specific target
sequences in the presence of a
fluorescently labeled detector
probe. |
9
| Item | Aptima HSV Assay
(Subject Device) | BD ProbeTec Assay
(Predicate Device) |
|---------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Platform | The Panther system is an in vitro
diagnostic device intended for use
in clinical laboratories by trained
laboratory professionals for theautomation of NAATs. The Panther
system provides automation of all
assay steps, including sample
processing, amplification of target
nucleic acid, amplicon detection,
data reduction, and amplicon
inactivation. | BD Viper System is an in vitro
diagnostic device intended for
use in clinical laboratories by
trained laboratory professionals
for the automation of NAATs.
The BD Viper System provides
automated non-specific
extraction and amplification of
DNA from clinical specimens.
In Extracted Mode, it uses
Strand Displacement
Amplification technology.
When present, HSV 1 and/or 2
amplicon is detected by Strand
Displacement Amplification. |
| Specimen Collection | • Aptima Multitest Swab
Specimen Collection Kit (STM)
• Aptima Specimen Transfer Kit
(for use with specimens collected
in VTM) | • BD ProbeTecQˣ Collection
Kit for Endocervical or
Lesion Specimens
• BD Universal Viral
Transport Medium (VTM)
w/Swab Collection Kit
Copan Universal Transport
Medium w/Swab Collection Kit
(same as BD VTM Kit) |
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 studies (non-clinical) were conducted to support the clearance of the Aptima HSV Assay on the Panther system.
Analytical Sensitivity and Limit of Detection (LoD)
The LoD in clinical specimen matrix was determined by testing dilution panels for each of two strains of HSV-1 virus (Macintyre and HF strains) and two strains of HSV-2 viral (MS and G) in
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both Aptima Specimen Transport Media (STM) and Viral Transport Media (VTM) negative clinical pools in the Aptima HSV assay. LoD for the assay was based on results from the reagent lot with the highest concentration for the 95% predicted detection limit. Table 3 contains the probit-predicted 95% LoD values of the Aptima HSV assay for HSV-1 and HSV-2 in both specimen types.
| HSV Type/Strain | Specimen Type | LoD
TCID50/mL |
|-----------------|---------------|------------------|
| HSV-1 MacIntyre | STM | 60.6 |
| HSV-1 MacIntyre | VTM | 186.9 |
| HSV-1 HF | STM | 78.9 |
| HSV-1 HF | VTM | 159.3 |
| HSV-2 MS | STM | 18.2 |
| HSV-2 MS | VTM | 28.7 |
| HSV-2 G | STM | 18.8 |
| HSV-2 G | VTM | 128.8 |
Table 3: HSV 1 & 2 LoD in VTM and STM
Limit of Detection Verification
LoD of the Aptima HSV 1 & 2 assay was verified using two clinical isolates of HSV-1 and two clinical isolates of HSV-2 from HSV-positive clinical specimens that were cultured and quantitated in-house. Each isolate was tested with the Aptima HSV 1 & 2 assay using 60 replicates each at 1X LoD, 3X LoD, and 10X LoD, see Table 4 for LoD values. Testing was completed in both STM and VTM matrix for all four clinical isolates and was conducted using 3 lots of reagents. All replicates for all clinical isolates at all three concentrations tested were detected by the Aptima HSV 1 & 2 assay, demonstrating that the assay can accurately detect a range of both HSV-1 and HSV-2 isolates at the determined LoD.
| HSV Type | Media Type | LoD (TCID50/mL) |
|---|---|---|
| HSV-1 | STM | 78.9 |
| VTM | 186.9 | |
| HSV-2 | STM | 18.8 |
| VTM | 128.8 |
Table 4: HSV 1 and HSV 2 LoD
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Limit of Detection in Neat STM.
The LoD in neat STM (Aptima Specimen Transport Media) was determined by testing panels that consisted of dilutions of HSV-1 and HSV-2 in vitro RNA transcripts (IVT) and dilutions of HSV-1 and HSV-2 viral particles in neat STM matrix. Through Probit analysis, results show the Aptima HSV Assay has a predicted 95% LoD of 68.3 copies/mL for HSV-1 IVT and 49.4 copies/mL for HSV-2 IVT. The assay has a predicted 95% LoD of 28.9 TCID%ML for HSV-1 MacIntyre virus and 0.54 TCID50/mL for HSV-2 MS virus.
Interfering Substances
Potentially interfering substances listed in Table 4 were tested in the Aptima HSV 1 & 2 assay. Panels were built in STM and evaluated for potential effects on both assay sensitivity and specificity. Sensitivity performance was evaluated separately for both HSV-1 and HSV-2 by spiking viral particles into substance-containing panels at 3X the LoD. HSV-negative panels containing each substance were also evaluated for specificity.
No effect on assay performance was observed in the presence of a representative brand of the following exogenous and endogenous substances tested at the concentrations indicated in Table 5.
| Interfering Substance | Final Concentration |
|---|---|
| Vaginal lubricant | 5% V/V |
| Spermicide/Contraceptive Jelly | 4% W/V |
| Anti-fungal cream | 5% W/V |
| Douche | 5% V/V |
| Feminine Spray | 5% W/V |
| Body Lotion | 5% W/V |
| Powder | 5% W/V |
| Glacial Acetic Acid Wash Solution | 5% V/V |
| Hemorrhoidal Cream | 5% W/V |
| Urine | 5% V/V |
| Whole Blood | 0.5% V/V |
| Leukocytes | $4x10^5$ Cells/mL |
| Mucus | 0.3% W/V |
| Seminal Fluid | 5% V/V |
| Feces | 0.03% W/V |
| Protein | 4% W/V |
| Antiviral drug (Acyclovir) | 5% W/V |
Table 5: Potential Interfering Substances
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Co-Infection
A co-infection study was evaluated to assess the ability of the Aptima HSV 1 & 2 assay to detect HSV 1 and HSV 2 analytes when both are present in one specimen. Low and high titer concentrations of HSV-1 viral particles in STM were tested in combination with low, moderate, and high concentrations of HSV-2 virus. Panel composition and concentrations are listed in Table 6. All testing resulted in 100% detection for both HSV-1 and HSV-2.
| Panel Member | HSV-1 Concentration | HSV-2 Concentration |
|---|---|---|
| HSV-1 Low; HSV-2 Low | 86.7 TCID50/mL1 | 1.62 TCID50/mL2 |
| HSV-1 Low; HSV-2 Moderate | 86.7 TCID50/mL1 | 540 TCID50/mL3 |
| HSV-1 High; HSV-2 Low | 28,900 TCID50/mL4 | 1.62 TCID50/mL2 |
| HSV-1 High; HSV-2 Moderate | 28,900 TCID50/mL4 | 540 TCID50/mL3 |
| HSV-1 Low; HSV-2 High | 86.7 TCID50/mL1 | 9,000 TCID50/mL5 |
Table 6: Co-infected Panel
¹ 3X LoD HSV-1.
2 3X LoD HSV-2.
3 1000X LoD HSV-2.
4 1000X LoD HSV-1.
5 16667X LoD HSV-2.
Cross-Reactivity
A study was performed to evaluate the cross-reactivity and interference with the Aptima HSV 1 & 2 assay in the presence of non-targeted microorganisms that could be present in clinical specimens. A panel consisting of 48 microorganisms (Table 7) was tested in STM at a test concentration of 1 x 105 units/mL for viruses and 1 x 106 units/mL for all other organisms. Organisms were tested in the absence of HSV or in the presence of either HSV-1 or HSV-2 at 3X LoD. Thirty replicates were tested per panel condition. No evidence of cross-reactivity or microbial interference was observed for any of the 48 test microorganisms tested with the Aptima HSV 1 & 2 assay, at the following concentrations.
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| Microorganism | Concentration |
|---|---|
| Acinetobacter calcoaceticus | 1x106 CFU/mL1,2 |
| Acinetobacter lwoffii | 1x106 CFU/mL1,2 |
| Actinomyces israelii | 1x106 RNA copies /mL2 |
| Adenovirus type 1 | 1x105 TCID50/mL3 |
| Alcaligenes faecalis | 1x106 CFU/mL1 |
| Atopobium vaginae | 1x106 RNA copies /mL2 |
| Bacteroides fragilis | 1x106 CFU/mL1,2 |
| Bifidobacterium adolescentis | 1x106 CFU/mL1,2 |
| BK virus | 1x105 DNA copies/mL3 |
| Bordetella bronchiseptica | 1x106 CFU/mL1,2 |
| Bordetella pertussis | 1x106 CFU/mL1,2 |
| Campylobacter jejuni | 1x106 CFU/mL1,2 |
| Candida glabrata | 1x106 CFU/mL1,2 |
| Clostridium difficile | 1x106 CFU/mL1,2 |
| Clostridium perfringens | 1x106 CFU/mL1,2 |
| Corynebacterium genitalium | 1x106 CFU/mL1,2 |
| Cryptococcus neoformans | 1x106 CFU/mL1,2 |
| Enterobacter aerogenes | 1x106 CFU/mL1,2 |
| Enterobacter cloacae | 1x106 CFU/mL1,2 |
| Enterococcus faecium | 1x106 CFU/mL1,2 |
| Enterococcus faecalis | 1x106 CFU/mL1,2 |
| Epstein-Barr virus | 1x105 DNA copies/mL3 |
| Escherichia coli | 1x106 CFU/mL1,2 |
| Fusobacterium nucleatum | 1x106 CFU/mL1,2 |
| Gardnerella vaginalis | 1x106 CFU/mL1,2 |
| Haemophilus ducreyi | 1x106 CFU/mL1,2 |
| Hepatitis B virus | 1x105 IU/mL4,3 |
| Klebsiella pneumoniae | 1x106 CFU/mL1,2 |
| Lactobacillus crispatus | 1x106 CFU/mL1,2 |
| Moraxella catarrhalis | 1x106 CFU/mL1,2 |
| Mycoplasma hominis | 1x106 RNA copies /mL2 |
| Mycoplasma orale | 1x106 RNA copies /mL2 |
| Neisseria gonorrhoeae | 1x106 CFU/mL1,2 |
| Neisseria meningitidis | 1x106 CFU/mL1,2 |
| Parvovirus B19 | 1x105 TCID50/mL3 |
| Prevotella bivia | 1x106 CFU/mL1,2 |
| Microorganism | Concentration |
| Propionibacterium acnes | 1x106 CFU/mL1,2 |
| Proteus mirabilis | 1x106 CFU/mL1,2 |
| Proteus vulgaris | 1x106 CFU/mL1,2 |
| Pseudomonas aeruginosa | 1x106 CFU/mL1,2 |
| Staphylococcus aureus | 1x106 CFU/mL1,2 |
| Staphylococcus epidermidis | 1x106 CFU/mL1,2 |
| Staphylococcus saprophyticus | 1x106 CFU/mL1,2 |
| Streptococcus mitis | 1x106 CFU/mL1,2 |
| Streptococcus pneumoniae* | 1x105 CFU/mL1,2 |
| Streptococcus pyogenes | 1x106 CFU/mL1,2 |
| Varicella-zoster virus | 1x105 DNA copies/mL3 |
| West Nile virus | 1x105 TCID50/mL3 |
Table 7: Cross-Reactivity and Microbial Interference Panel
14
Table 7: Cross-Reactivity and Microbial Interference Panel
1 CFU = Colony Forming Units, 2Procured internally from Hologic, Inc., 3 Obtained from ZeptoMetrix Corporation (Buffalo NY), 4IU=International Units *Cross reactivity was observed in Streptococcus pneumoniae at 1x106 CFU/mL
Specimen Inhibition in Clinical Samples
An evaluation was conducted of the whole system failure rate leading to false negative or invalid results by testing low positive specimens. HSV negative VTM and STM samples were spiked with HSV-1 Macintyre or HSV-2 MS strain cultured virus at a test concentration at 3X LoD. Of 201 STM specimens tested, the false negative rate was 0% (0/201; 95% CI: 0.0-1.9) for HSV-1 and 0.5% (1/201; 95% CI: 0.1-2.8) for HSV-2. Of 202 VTM specimens tested, the false negative rate was 0% (0/202; 95% CI: 0.0-1.9) for HSV-1 and 0% (0/202; 95% CI: 0.0-1.9) for HSV-2.
Reproducibility and Within Laboratory Precision
To evaluate the reproducibility and within laboratory precision of the Aptima HSV assay, one HSV negative panel and six HSV positive panels were tested. The six HSV positive panels were spiked with HSV-1 cultured virus strain (MacIntyre) and/or HSV-2 cultured virus strain (MS). All panel members were made using STM. Three replicates of each panel was tested by three operators in two runs each in three reagent lots over 13 days.
15
Results showed that the Aptima HSV assay has acceptable precision and is reproducible with various instruments, reagent lots, and operators.
Brief Description of Clinical Studies
Clinical testing of the Aptima HSV assay on the Panther system included performance and reproducibility testing. Substantial equivalence is based in part on the performance study.
Reproducibility
Aptima HSV 1 & 2 assay reproducibility was evaluated at three external US sites. Testing was performed using three lots of assay reagents and six operators (two at each site). At each site, testing was performed for at least six days. Panel members were created by spiking HSV-1 and/or HSV-2 viral particles into STM. Final HSV-1 concentrations ranged from 0 TCID50mL to 86.96 TCID50/mL and final HSV-2 concentrations ranged from 0 TCID50/mL to 1.63 TCID50/mL.
HSV-negative panel members and panel members containing low (99.9) | 98.6
(93.0-100) | 98.9
(97.6-99.6) |
| | Male | 192 | 19 | 1 | 170 | 2 | 10.9 | 90.5
(71.1-97.3) | 99.4
(96.8-99.9) | 95.0
(78.6-99.8) | 98.8
(96.4-99.9) |
| | Female | 336 | 52 | 0 | 281 | 3 | 16.4 | 94.5
(85.1-98.1) | 100
(98.7-100) | 100
(93.7-100) | 98.9
(97.1-99.8) |
| STM | Combined | 531 | 71 | 25 | 454 | 46 | 14.1 | 94.7
(87.1-97.9) | 99.6
(98.4-99.9) | 97.3
(91.1-99.6) | 99.1
(97.9-99.8) |
| | Male | 192 | 20 | 2 | 169 | 1 | 10.9 | 95.2
(77.3-99.2) | 98.8
(95.8-99.7) | 90.9
(74.5-98.7) | 99.4
(97.2-100) |
| | Female | 339 | 51 | 0 | 285 | 3 | 15.9 | 94.4
(84.9-98.1) | 100
(98.7-100) | 100
(93.6-100) | 99.0
(97.2-99.8) |
Table 11. Summary of HSV-1 Results by VTM and STM Specimen Type
STM = Aptima Multitest Swab specimen, Prev = prevalence, VTM = VTM sample
1Score CI
2PPV 95% CI computed from the exact 95% CI for the positive likelihood ratio, NPV 95% CI computed from the exact 95% CI for the negative likelihood ratio
3The sample had a negative culture result.
4Two samples had negative culture results, one had a non-typable HSV positive culture result, and two were HSV-1 positive by culture. 5Both specimens had negative culture results.
6 One specimen had a negative culture result, one had a non-typable HSV positive culture result, and two were HSV-1 positive by culture.
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| Specimen
Type | Gender | N | TP | FP | TN | FN | Prev
(%) | Sensitivity %
(95% CI)1 | Specificity %
(95% CI)1 | PPV %
(95% CI)2 | NPV %
(95% CI)2 |
|------------------|----------|-----|-----|-----|-----|----|-------------|----------------------------|----------------------------|---------------------|---------------------|
| VTM | Combined | 533 | 248 | 73 | 270 | 84 | 48.0 | 96.9
(94.0-98.4) | 97.5
(94.9-98.8) | 97.3
(94.7-98.8) | 97.1
(94.6-98.7) |
| | Male | 194 | 79 | 2 | 110 | 3 | 42.3 | 96.3
(89.8-98.7) | 98.2
(93.7-99.5) | 97.5
(92.0-99.7) | 97.3
(93.0-99.4) |
| | Female | 339 | 169 | 5 | 160 | 5 | 51.3 | 97.1
(93.5-98.8) | 97.0
(93.1-98.7) | 97.1
(93.8-99.0) | 97.0
(93.4-99.0) |
| STM | Combined | 535 | 253 | 205 | 258 | 46 | 48.0 | 98.4
(96.1-99.4) | 92.8
(89.1-95.3) | 92.7
(89.4-95.3) | 98.5
(96.3-99.6) |
| | Male | 194 | 79 | 6 | 106 | 3 | 42.3 | 96.3
(89.8-98.7) | 94.6
(88.8-97.5) | 92.9
(86.5-97.1) | 97.2
(92.8-99.4) |
| | Female | 341 | 174 | 14 | 152 | 1 | 51.3 | 99.4
(96.8-99.9) | 91.6
(86.3-94.9) | 92.6
(88.5-95.7) | 99.3
(96.6-100) |
Table 12. Summary of HSV-2 Results by VTM and STM Specimen Type
STM = Aptima Multitest swab specimen, Prev = prevalence, VTM = VTM sample
1Score CI
2PV 95% CI computed from the exact 95% CI for the positive likelihood ratio, NPV 95% CI computed from the exact 95% CI for the negative likelihood ratio
3Six samples had negative culture results and one was HSV-1 positive by culture.
4All eight samples had negative culture results.
5Eighteen specimens had negative culture results and two were HSV-1 positive by culture.
6All four specimens had negative culture results.
Expected Values
Prevalence
The prevalence of HSV-1 and HSV-2 observed during a multi-center clinical study was calculated for the Aptima HSV 1 & 2 assay. The prevalence of HSV-1 and HSV-2 with the Aptima HSV 1 & 2 assay is summarized by age group, gender group, and specimen type in Table 13.
21
| Gender | Age Group | VTM | STM | ||
|---|---|---|---|---|---|
| HSV-1 | HSV-2 | HSV-1 | HSV-2 | ||
| Male | All ages | 10.4 (20/192) | 41.8 (81/194) | 11.5 (22/192) | 43.8 (85/194) |
| 60 years | 0.0 (0/6) | 50.0 (3/6) | 0.0 (0/6) | 66.7 (4/6) | |
| Female | All ages | 15.5 (52/336) | 51.3 (174/339) | 15.0 (51/339) | 55.1 (188/341) |
| 60 years | 0.0 (0/3) | 100 (4/4) | 0.0 (0/3) | 100 (4/4) | |
| Combined | All ages | 13.6 (72/528) | 47.8 (255/533) | 13.7 (73/531) | 51.0 (273/535) |
| 60 years | 0.0 (0/9) | 70.0 (7/10) | 0.0 (0/9) | 80.0 (8/10) |
Table 13. Gender and Age Distribution by VTM and STM Specimen Type¹
1 No subjects had positive Aptima HSV 1 & 2 assay results for both HSV-1 and HSV-2.
22
Positive and Negative Predictive Values for Hypothetical Prevalence Rates
The estimated positive and negative predictive values (PPV and NPV) of the Aptima HSV 1 & 2 assay for detection of HSV-1 and HSV-2 across different hypothetical prevalence rates are shown for each specimen type in Table 14. These calculations are based on the overall estimated sensitivity and specificity for each specimen type as determined in the clinical performance study.
| HSV-1 | HSV-2 | ||||
|---|---|---|---|---|---|
| Prevalence (%) | PPV (%) | NPV (%) | PPV (%) | NPV (%) | |
| Specimen Type | |||||
| VTM | 1 | 81.0 | 99.9 | 27.9 | 100 |
| 2 | 89.6 | 99.9 | 43.9 | 99.9 | |
| 5 | 95.7 | 99.7 | 66.9 | 99.8 | |
| 10 | 97.9 | 99.3 | 81.0 | 99.6 | |
| 20 | 99.1 | 98.4 | 90.6 | 99.2 | |
| 30 | 99.5 | 97.3 | 94.3 | 98.6 | |
| 40 | 99.6 | 95.8 | 96.2 | 97.9 | |
| 50 | 99.8 | 93.8 | 97.5 | 96.9 | |
| STM | 1 | 68.6 | 99.9 | 12.1 | 100 |
| 2 | 81.5 | 99.9 | 21.8 | 100 | |
| 5 | 91.9 | 99.7 | 41.9 | 99.9 | |
| 10 | 96.0 | 99.4 | 60.3 | 99.8 | |
| 20 | 98.2 | 98.7 | 77.4 | 99.6 | |
| 30 | 98.9 | 97.8 | 85.4 | 99.3 | |
| 40 | 99.3 | 96.6 | 90.1 | 98.9 | |
| 50 | 99.5 | 94.9 | 93.2 | 98.4 |
Table 14. Prevalence vs Hypothetical PPV1 and NPV2 for Detection of HSV-1 and HSV-2 by Specimen Type
STM = Aptima Multitest swab specimen, VTM = VTM sample
1PPV was calculated using:
(Sensitivity x Prevalence) / (Sensitivity x Prevalence + [1 - Specificity] x [1 - Prevalence]).
2NPV was calculated using:
(Specificity x [1 - Prevalence]) / ([1 - Sensitivity] x Prevalence + Specificity x [1 - Prevalence]).
23
TTime Distribution for Aptima HSV 1 & 2 Assay Positive Controls
The distribution of the TTime values for the Aptima HSV 1 & 2 assay positive control from all valid Aptima HSV 1 & 2 assay runs performed during the clinical performance study is presented in Table 15.
| TTime | ||
|---|---|---|
| Statistics | HSV-1 | HSV-2 |
| N | 107 | 107 |
| Mean | 20.03 | 22.01 |
| Median | 19.8 | 21.7 |
| SD | 1.198 | 1.612 |
| CV (%) | 6.0 | 7.3 |
| Minimum | 18.1 | 19.5 |
| Maximum | 22.9 | 26.2 |
Table 15. Distribution of TTimes for Aptima HSV 1 & 2 Assay Positive Controls
CV = coefficient of variation, SD = standard deviation
VIII. CONCLUSIONS
The analytical and clinical study results demonstrate that the Aptima HSV assay on the Panther system is substantially equivalent to the predicate device that is currently marketed for the same intended use. Hardware and software verification and validation demonstrate that the Aptima HSV assay on the Panther system will perform as intended.