artus HSV-1/2 QS-RGQ MDx Kit
Device Facts
| Record ID | K142738 |
|---|---|
| Device Name | artus HSV-1/2 QS-RGQ MDx Kit |
| Applicant | Qiagen |
| Product Code | OQO · Microbiology |
| Decision Date | Dec 19, 2014 |
| Decision | SESE |
| Submission Type | Traditional |
| Regulation | 21 CFR 866.3305 |
| Device Class | Class 2 |
| Attributes | Pediatric |
Indications for Use
The artus HSV-1/2 QS-RGQ MDx Kit is an in vitro real-time PCR DNA amplification assay performed on the QIAsymphony RGQ MDx system for the direct qualitative detection and differentiation of herpes simplex virus (HSV-1 and HSV-2) DNA in genital or oral vesicular lesions from male and female patients suspected of HSV infection. The assay is intended for use as an aid in diagnosis of HSV infection in symptomatic patients. Warning: The artus HSV-1/2 QS-RGQ MDx Kit is not FDA-cleared for use with cerebrospinal fluid (CSF) or for prenatal screening.
Device Story
The artus HSV-1/2 QS-RGQ MDx Kit is an in vitro real-time PCR assay for qualitative detection and differentiation of HSV-1 and HSV-2 DNA. Input samples consist of genital or oral lesion swabs. The workflow utilizes the QIAsymphony SP/AS instrument for automated DNA extraction and PCR setup, followed by amplification and detection on the Rotor-Gene Q MDx (RGQ MDx) instrument. The assay uses fluorescently labeled oligonucleotide probes; signal generation is inversely proportional to target concentration. An internal control (synthetic plasmid) is added during sample preparation to monitor extraction and PCR inhibition. The Rotor-Gene AssayManager software automatically analyzes results, reporting 'Signal detected', 'No signal', or 'INVALID'. The device is intended for use in clinical laboratories by trained personnel. Results aid clinicians in diagnosing HSV infection in symptomatic patients.
Clinical Evidence
Prospective clinical study of 662 samples (510 genital, 152 oral) compared the device to ELVIS HSV ID and D3 Typing Test System. For genital samples (N=414 for HSV-1, N=510 for HSV-2), HSV-1 sensitivity was 95.8% and specificity 93.9%; HSV-2 sensitivity was 96.9% and specificity 91.1%. For oral samples (N=152), HSV-1 sensitivity was 93.8% and specificity 81.7%. Discordant analysis using alternative PCR and sequencing supported the device results. Contrived sample studies provided additional data for HSV-2 in oral samples.
Technological Characteristics
Real-time PCR assay; utilizes fluorescently labeled oligonucleotide probes for target detection. Instrumentation: QIAsymphony SP/AS for extraction/setup; Rotor-Gene Q MDx for amplification/detection. Software: Rotor-Gene AssayManager. Internal control: synthetic plasmid DNA. Storage: -30°C to -10°C. Connectivity: Integrated system workflow.
Indications for Use
Indicated for symptomatic male and female patients suspected of HSV infection. Used for qualitative detection and differentiation of HSV-1 and HSV-2 DNA in genital or oral vesicular lesions. Not indicated for CSF or prenatal screening.
Regulatory Classification
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.
Special Controls
*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).
Predicate Devices
- MultiCode®-RTx Herpes Simplex Virus 1 & 2 Kit (K100336)
Related Devices
- K150962 — Simplexa HSV 1 & 2 Direct, Simplexa HSV 1 & 2 Positive Control Pack · Focus Diagnostics · Aug 28, 2015
- K142156 — SEEGENE ANYPLEX II HSV-1/2 ASSAY · Seegene · Feb 13, 2015
Submission Summary (Full Text)
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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY
A. 510(k) Number:
K142738
B. Purpose for Submission:
New device
C. Measurand:
Target DNA sequences from Herpes Simplex Virus type 1 (HSV-1) and Herpes Simplex Virus type 2 (HSV-2)
D. Type of Test:
Real-time PCR DNA amplification assay
E. Applicant:
QIAGEN
F. Proprietary and Established Names:
artus® HSV-1/2 QS-RGQ MDx Kit
G. Regulatory Information:
1. Regulation section:
21 CFR 866.3305
2. Classification:
Class II
3. Product code:
OQO
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4. Panel:
Microbiology (83)
H. Intended Use:
1. Intended use(s):
The artus HSV-1/2 QS-RGQ MDx Kit is an in vitro real-time PCR DNA amplification assay performed on the QIAsymphony RGQ MDx system for the direct qualitative detection and differentiation of herpes simplex virus (HSV-1 and HSV-2) DNA in genital or oral vesicular lesions from male and female patients suspected of HSV infection.
The assay is intended for use as an aid in diagnosis of HSV infection in symptomatic patients.
Warning: The artus HSV-1/2 QS-RGQ MDx Kit is not FDA-cleared for use with cerebrospinal fluid (CSF) or for prenatal screening.
2. Indication(s) for use:
Same as Intended Use
3. Special conditions for use statement(s):
For prescription use only
4. Special instrument requirements:
The artus HSV-1/2 QS-RGQ MDx Kit is performed on the QIAsymphony RGQ MDx system
I. Device Description:
The artus HSV-1/2 QS-RGQ MDx Kit is an in vitro PCR assay for the qualitative detection and differentiation of nucleic acids encoding the Glycoprotein D and UL30 genes isolated from HSV-1 and HSV-2 DNA present in genital or oral lesions from male and female patients. Samples are extracted and prepared for PCR using the QIAsymphony SP/AS instrument with the QIAsymphony DSP Virus/Pathogen Mini Kit. Amplification and detection are carried out using the artus HSV-1/2 QS-RGQ MDx Kit with the Rotor-Gene Q MDx (RGQ MDx) and Rotor-Gene AssayManager software. The presence of a HSV-1 or HSV-2 target sequence is indicated by the fluorescent signal generated through the use of fluorescently labeled oligonucleotide probes. The probes do not generate a signal unless they are specifically bound to the amplified product. The amplification cycle at which fluorescent signal is detected by the RGQ MDx is inversely proportional to the HSV-1 and/or HSV-2 target concentration present in the original specimen. A plasmid construct containing DNA
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unrelated to HSV-1 and HSV-2 is introduced into each specimen during sample preparation to serve as an internal control. Run as a separate control, the positive control serves to demonstrate that the HSV-1/2 PCR reagents are functional. In addition, the positive control functions as a process control, to demonstrate that sample preparation has proceeded correctly during the run.
# J. Substantial Equivalence Information:
1. Predicate device name(s):
MultiCode®-RTx Herpes Simplex Virus 1 & 2 Kit (Eragen)
2. Predicate 510(k) number(s):
K100336
3. Comparison with predicate:
| Similarities | | |
| --- | --- | --- |
| Item | Device | Predicate |
| Indications for use | The artus HSV-1/2 QS-RGQ MDx Kit is an in vitro real-time PCR DNA amplification assay performed on the QIAsymphony RGQ MDx system for the direct qualitative detection and differentiation of herpes simplex virus (HSV-1 and HSV-2) DNA in genital or oral vesicular lesions from male and female patients suspected of HSV infection.The assay is intended for use as an aid in diagnosis of HSV infection in symptomatic patients.Warning: The artus HSV-1/2 QS-RGQ MDx Kit is not FDA-cleared for use with cerebrospinal fluid (CSF) or for prenatal screening. | The MultiCode®-RTx Herpes Simplex Virus 1 & 2 Kit is a polymerase chain reaction (PCR)-based qualitative in vitro diagnostic test for the detection and typing of herpes simplex virus (HSV1&2) DNA in vaginal lesions. It is indicated for use in the detection and typing of HSV-1 or HSV-2 in vaginal lesion swab specimens from symptomatic female patients as an aid in the diagnosis of genital herpes infection.Warning: The device is not FDA cleared for the use with cerebral spinal fluid (CSF) or any lesions other than vaginal. The assay is not intended to be used for |
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| Similarities | | |
| --- | --- | --- |
| Item | Device | Predicate |
| | | male penile specimens, for prenatal screening, or females under the age of 18 years. |
| Technology | Real-time PCR | same |
| Target detected | HSV-1 and HSV-2 DNA | same |
| Differences | | |
| --- | --- | --- |
| Item | Device | Predicate |
| Sample type | Male and female genital or oral herpetic lesions | Female vaginal lesions |
# K. Standard/Guidance Document Referenced (if applicable):
Not Applicable
# L. Test Principle:
The HSV-1/2 Master A and HSV-1/2 Master B components contain reagents and enzymes for the specific amplification of target regions within the HSV-1 and HSV-2 genomes and for the direct detection of the specific amplicon in fluorescence channels Cycling Orange and Cycling Green respectively of the Rotor-Gene Q MDx.
In addition, the artus HSV-1/2 QS-RGQ MDx Kit contains a second heterologous control system to identify potential failures during the assay process. This is detected as an internal control (IC) in fluorescence channel Cycling Crimson of the Rotor-Gene Q MDx.
The contents of the artus HSV-1/2 QS-RGQ MDx Kit are sufficient for 72 tests in one to 3 batches of 24 reactions on the QIAsymphony RGQ MDx. The Rotor-Gene Q MDx rotor holds up to 72 reaction tubes.
Kit contents
| artus HSV-1/2 QS-RGQ MDx Kit | | (72) |
| --- | --- | --- |
| Catalog no. | | 4526346 |
| Number of reactions | | 72 |
| Blue | HSV-1/2 Master A | 3 x 330 μl |
| Violet | HSV-1/2 Master B | 3 x 600 μl |
| Green | HSV-1/2 Internal Control | 3 x 540 μl |
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| Red | HSV-1/2 Positive Control | | 3 x 330 μl |
| --- | --- | --- | --- |
| White | HSV-1/2 Negative Control | | 3 x 350 μl |
| artus HSV-1/2 QS-RGQ MDx Kit Instructions for Use (Handbook) | | | 1 |
## Materials Required but Not Provided
The following is a list of some of the major materials required but not provided with the kit.
1. Equipment for sample preparation and assay setup:
- QIAsymphony SP (module of the QIAsymphony RGQ MDx)*
- QIAsymphony AS (module of the QIAsymphony RGQ MDx)*
- QIAsymphony software version 4.0
2. Equipment for PCR
- Rotor-Gene Q MDx (module of the QIAsymphony RGQ MDx)*
- Rotor-Gene AssayManager® version 1.0 (module of the QIAsymphony RGQ MDx)
## Interpretation of Results
The artus HSV-1/2 QS-RGQ MDx Kit Assay Profile automatically analyzes samples, positive and negative controls, and run results. Every sample and control displays an independent result for each target: HSV-1, HSV-2, and Internal Control. Each result is reported as "Signal detected", "No signal", or "INVALID".
All targets for the positive control and negative control must be valid to confirm that the assay status is successful and that the test results may be reported. If any target of the positive control or negative control is invalid, results for every sample in the run will display "INVALID"; the entire assay run must then be retested.
The positive control must report a "Signal detected" result for HSV-1, HSV-2, and Internal Control. The negative control must report a "Signal detected" result for Internal Control and "No signal" for the specified HSV-1 and HSV-2 target.
It is expected that in some positive HSV-1/2 samples the Internal Control PCR may be inhibited due to competition from amplifying HSV-1/2, which will cause a "No signal" or "INVALID" result for Internal Control.
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Summary of results interpretation
| Target result | | | | HSV detected in sample |
| --- | --- | --- | --- | --- |
| HSV-1 | HSV-2 | Internal Control | Sample status | |
| Signal detected | Signal detected | Signal detected/ No signal/ INVALID | Valid | Yes |
| Signal detected | No signal | Signal detected/ No signal/ INVALID | Valid | Yes |
| No signal | Signal detected | Signal detected/ No signal/ INVALID | Valid | Yes |
| No signal | No signal | Signal detected | Valid | No |
| No signal | No signal | No signal/ INVALID | Invalid | Error, retest sample |
| Signal detected | INVALID | Signal detected/ No signal/ INVALID | Valid | Yes |
| INVALID | Signal detected | Signal detected/ No signal/ INVALID | Valid | Yes |
| INVALID | No signal | No signal/ INVALID | Invalid | Error, retest sample |
| No signal | INVALID | No signal/ INVALID | Invalid | Error, retest sample |
| No signal | INVALID* | Signal detected | Valid | Error, retest sample |
| INVALID* | No signal | Signal detected | Valid | Error, retest sample |
*If a target is reported as “INVALID” and the flag says CT_ABOVE_ACCEPTED_RANGE, this sample does not need to be retested and is considered “No signal”, if the Internal Control is valid.
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M. Performance Characteristics (if/when applicable):
1. Analytical performance:
a. Precision/Reproducibility:
The precision of the artus HSV-1/2 QS-RGQ MDx Kit was assessed using a seven-member precision panel consisting of 2 strains of HSV: HSV-1 MacIntyre and HSV-2 MS, diluted in M4RT viral transport medium. Panel members were formulated with a single HSV strain present (HSV-1 or HSV-2) at three concentrations; Positive (~2-3X LoD), Low Positive (1X LoD), and High Negative (<1X LoD, targeting a 20-80% positivity rate). A seventh panel member (Negative) was prepared using M4RT viral transport medium only. The data obtained were used to determine the mean $C_T$, standard deviation (ST DEV) and the coefficient of variation (%CV) for each target and the internal control.
For the within laboratory repeatability study, the seven-member panel was tested in replicates of three, once a day for a total of twelve days. The testing was conducted by two alternating operators using one QIAsymphony RGQ MDx (QS-RGQ MDx) instrument platform and one reagent kit lot.
Within-Laboratory Repeatability Study Results
| Sample | HSV-1 | | | HSV-2 | | | IC | | | Detected/Total |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | Mean $C_T$ | ST DEV | %CV | Mean $C_T$ | ST DEV | %CV | Mean $C_T$ | ST DEV | %CV | |
| HSV-1 Positive | 32.56 | 0.32 | 0.97% | N/A | N/A | N/A | 31.79 | 0.64 | 2.00% | 35/35* |
| HSV-1 Low Positive | 33.89 | 0.35 | 1.03% | N/A | N/A | N/A | 31.85 | 0.66 | 2.07% | 36/36 |
| HSV-1 High Negative | 37.52 | 0.63 | 1.69% | N/A | N/A | N/A | 31.73 | 0.59 | 1.87% | 23/36 |
| HSV-2 Positive | N/A | N/A | N/A | 33.73 | 0.41 | 1.22% | 31.72 | 0.63 | 1.98% | 36/36 |
| HSV-2 Low Positive | N/A | N/A | N/A | 36.73 | 1.05 | 2.86% | 31.69 | 0.63 | 2.00% | 36/36 |
| HSV-2 High Negative | N/A | N/A | N/A | 38.35 | 1.01 | 2.64% | 31.82 | 0.57 | 1.80% | 9/36 |
| HSV Negative | N/A | N/A | N/A | N/A | N/A | N/A | 31.88 | 0.52 | 1.64% | 0/36 |
* Total number of samples is less than 36 due to exclusion of 1 invalid sample
To measure site-to-site reproducibility, the 7-member panel was run by 2 users at each of 3 external sites. Each of the 2 users performed 5 runs on alternating testing days for a total of 90 test results per panel member. Panel members were tested in replicates of 3 that were randomized and blinded to the user. A single QIAsymphony RGQ MDx instrument platform and one lot of the artus HSV-1/2 QS-RGQ MDx Kit were used at each site to conduct the study.
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Site-to-Site Reproducibility Study Results
| Panel member | Site | HSV-1/HSV-2 | | | IC | | | Detected/Total |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | Mean CT | ST DEV | %CV | Mean CT | ST DEV | %CV | |
| HSV-1 Positive | Site 1 | 31.40 | 1.04 | 3.30% | 30.87 | 0.35 | 1.12% | 27/27 |
| | Site 2 | 31.44 | 0.35 | 1.10% | 30.80 | 0.37 | 1.19% | 29/29 |
| | Site 3 | 31.20 | 0.32 | 1.03% | 30.45 | 0.30 | 0.98% | 26/26 |
| | Overall | 31.35 | 0.66 | 2.09% | 30.71 | 0.38 | 1.24% | 82/82* |
| HSV-1 Low Positive | Site 1 | 33.01 | 0.40 | 1.20% | 30.91 | 0.34 | 1.11% | 30/30 |
| | Site 2 | 32.78 | 0.44 | 1.34% | 30.76 | 0.38 | 1.24% | 30/30 |
| | Site 3 | 32.37 | 0.89 | 2.74% | 30.47 | 0.35 | 1.13% | 30/30 |
| | Overall | 32.72 | 0.66 | 2.03% | 30.71 | 0.40 | 1.29% | 90/90 |
| HSV-1 High Negative | Site 1 | 36.34 | 0.88 | 2.41% | 30.85 | 0.29 | 0.93% | 19/30 |
| | Site 2 | 36.27 | 0.74 | 2.05% | 30.67 | 0.40 | 1.29% | 15/30 |
| | Site 3 | 36.77 | 1.06 | 2.88% | 30.44 | 0.30 | 1.00% | 17/30 |
| | Overall | 36.46 | 0.92 | 2.51% | 30.66 | 0.37 | 1.20% | 51/90 |
| HSV-2 Positive | Site 1 | 33.19 | 0.36 | 1.08% | 30.89 | 0.30 | 0.99% | 30/30 |
| | Site 2 | 32.85 | 0.35 | 1.05% | 30.75 | 0.35 | 1.15% | 30/30 |
| | Site 3 | 32.46 | 0.33 | 1.03% | 30.47 | 0.29 | 0.95% | 30/30 |
| | Overall | 32.83 | 0.45 | 1.38% | 30.70 | 0.36 | 1.17% | 90/90 |
| HSV-2 Low Positive | Site 1 | 36.12 | 0.96 | 2.65% | 30.84 | 0.29 | 0.95% | 30/30 |
| | Site 2 | 35.53 | 0.66 | 1.85% | 30.73 | 0.32 | 1.04% | 30/30 |
| | Site 3 | 35.81 | 0.92 | 2.58% | 30.49 | 0.30 | 0.97% | 30/30 |
| | Overall | 35.82 | 0.88 | 2.46% | 30.69 | 0.33 | 1.08% | 90/90 |
| HSV-2 High Negative | Site 1 | 38.09 | 0.55 | 1.44% | 30.91 | 0.31 | 0.99% | 5/30 |
| | Site 2 | 37.69 | 2.30 | 6.09% | 30.71 | 0.37 | 1.20% | 5/30 |
| | Site 3 | 37.27 | N/A | N/A | 30.50 | 0.32 | 1.04% | 1/30 |
| | Overall | 37.83 | 1.52 | 4.01% | 30.71 | 0.37 | 1.20% | 11/90 |
| HSV Negative | Site 1 | N/A | N/A | N/A | 30.88 | 0.30 | 0.96% | 0/30 |
| | Site 2 | N/A | N/A | N/A | 30.72 | 0.36 | 1.16% | 0/30 |
| | Site 3 | N/A | N/A | N/A | 30.53 | 0.32 | 1.04% | 0/30 |
| | Overall | N/A | N/A | N/A | 30.71 | 0.35 | 1.14% | 0/90 |
* Total number of samples is less than 90 due to exclusion of invalid samples
b. Linearity/assay reportable range:
Not applicable
c. Traceability, Stability, Expected values (controls, calibrators, or methods):
Traceability: Not applicable
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# Stability studies:
## On-Board Reagent Stability
Two HSV strains (HSV-1, McIntyre strain and HSV-2, MS strain from Zeptometrix Corporation (ZMC) were diluted to 2-3X LoD in M4RT viral transport medium. The samples and reagents were incubated on-board the QIAsymphony SP/AS for 65 minutes before initiating the QIAsymphony SP/AS run. Following completion of the QIAsymphony AS run, assembled PCRs were held in the instrument for an additional 65 minutes before transferring the assembled reactions to the RGQ MDx instrument and initiating PCR. All 35 replicates were detected for both HSV-1 and HSV-2 after 65 minutes of incubation before initiating the QIAsymphony SP/AS run followed by 65 minutes of incubation before initiating the PCR reaction in the RGQ MDx instrument.
## Sample Stability
One HSV-1 Isolate #15 strain and one HSV-2 MS strain, both from ZMC, were used to prepare contrived positive samples to assess specimen stability. Both strains were prepared in M4RT viral transport medium plus matrix at 2-3X LoD (54.6 TCID $_{50}$ /mL for HSV-1 Isolate #15 and 2.94 TCID $_{50}$ /mL for HSV-2 MS Strain) and $\geq$10X LoD (190 TCID $_{50}$ /mL for HSV-1 Isolate #15 and 12 TCID $_{50}$ /mL for HSV-2 MS Strain). Contrived negative specimens were also prepared, consisting of M4RT viral transport medium plus matrix alone. Six replicates of each strain at 2-3X LoD were stored at $2^{\circ}$C to $8^{\circ}$C, $-15^{\circ}$C to $-10^{\circ}$C, and $-30^{\circ}$C to $-20^{\circ}$C. Three replicates of each strain at $\geq$10X LoD as well as the negative specimen were also stored at the stated temperatures. Samples were removed at defined time intervals for testing. Testing was performed with samples stored at both ends of the temperature range (-15°C to -10°C and -30°C to -20°C) to support a specimen stability claim at -30°C to -10°C. A specimen was considered to be stable for a given time point/condition if the HSV-1 or HSV-2 target was detected for each positive sample replicate tested. The data from this study supports the HSV-1/2 specimen stability for 30 days at -30°C to -10°C and 7 days at $4^{\circ}$C and up to three freeze-thaw cycles in M4RT.
## Transport Stability
The artus HSV-1/2 QS-RGQ MDx Kit is a frozen product, stored at $-30^{\circ}\mathrm{C}$ to $-10^{\circ}\mathrm{C}$ and shipped on dry ice. The storage of the artus HSV-1/2 QS-RGQ MDx Kit at alternating temperatures for different time intervals was tested to simulate storage and transport conditions on dry ice for 72 hours. Individual kit lot materials were cycled from required storage conditions ($-30^{\circ}\mathrm{C}$ to $-10^{\circ}\mathrm{C}$ freezer) to shipping conditions ($-90^{\circ}\mathrm{C}$ to $-70^{\circ}\mathrm{C}$ freezer) and back to required storage conditions ($-30^{\circ}\mathrm{C}$ to $-10^{\circ}\mathrm{C}$ freezer), before testing the kit reagents. The artus HSV-1/2 QS-RGQ MDx Kit was tested at Day 0 in order to establish baseline performance. All replicates in each run (following 1X, 2X and 3X temperature cycles) passed stability testing.
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Controls:
# HSV-1/2 Internal Control:
The internal control consists of synthetic plasmid DNA. The Internal Control is input into the lysis buffer at a low concentration so that it is susceptible to the presence of potential inhibitors present in the sample matrix. Its function is to monitor the integrity of PCR reagents as well as the integrity of the PCR process without interfering with the target specific PCR.
# External controls:
HSV-1/2 positive and negative controls are loaded onto the QIAsymphony SP before DNA purification in place of a patient sample. They monitor the efficiency of sample preparation and assist in identifying contamination during sample preparation and/or the downstream assay. Positive Control should always be positive for HSV-1 and HSV-2 targets. Negative control should always be negative for HSV-1 and HSV-2 targets. However, the negative control may exhibit a signal in some cases but will still be considered negative as long as the signal obtained is outside the negative control $C_T$ cutoff.
# d. Detection limit:
# Analytical Sensitivity (Limit of Detection):
The limit of detection (LoD) was assessed for the artus HSV-1/2 QS-RGQ MDx Kit using two strains of HSV-1 (MacIntyre and Isolate #15 from ZMC) and two strains of HSV-2 (MS and Isolate #2 from ZMC). The LoD is defined as the HSV titer $(\mathrm{TCID}_{50} / \mathrm{mL})$ detected with a probability of $95\%$ or greater and was determined by probit analysis. The results, representative of the analytical sensitivity of the artus HSV-1/2 QS-RGQ MDx Kit, are summarized in the following table.
Limit of Detection
| Strain | LoD (95 % CI) TCID50/mL |
| --- | --- |
| HSV-1 Macintyre | 4.42 x 10^0 (2.81 x 10^0 - 9.14 x 10^0) |
| HSV-1 Isolate 15 | 1.82 x 10^1 (0.96 x 10^1 - 5.47 x 10^1) |
| HSV-2 MS | 9.78 x 10^-1 (6.6 x 10^-1 - 2.01 x 10^0) |
| HSV-2 Isolate 2 | 1.91 x 10^2 (1.26 x 10^2 - 3.55 x 10^2) |
# Analytical Reactivity:
The analytical reactivity of the artus HSV-1/2 QS-RGQ MDx Kit was assessed to determine whether the kit could detect a broad range of HSV-1 and HSV-2 strains. Strains were obtained from ZMC. A total of 39 strains (20 HSV-1 and 19 HSV-2) were diluted in M4RT viral transport medium to 2–3X LoD and tested with the artus HSV-1/2 QS-RGQ MDx Kit. The intended HSV-1 or HSV-2 was detected in all strains tested.
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Strains Tested in Analytical Reactivity
| Organism | Part No. | Organism | Part No. |
| --- | --- | --- | --- |
| HSV-1, Isolate #2 | 0810183CF | HSV-2, Isolate #3 | 0810204CF |
| HSV-1, Isolate #3 | 0810184CF | HSV-2, Isolate #4 | 0810205CF |
| HSV-1, Isolate #4 | 0810185CF | HSV-2, Isolate #5 | 0810206CF |
| HSV-1, Isolate #5 | 0810186CF | HSV-2, Isolate #6 | 0810207CF |
| HSV-1, Isolate #6 | 0810187CF | HSV-2, Isolate #7 | 0810208CF |
| HSV-1, Isolate #7 | 0810188CF | HSV-2, Isolate #8 | 0810209CF |
| HSV-1, Isolate #8 | 0810189CF | HSV-2, Isolate #9 | 0810210CF |
| HSV-1, Isolate #9 | 0810190CF | HSV-2, Isolate #10 | 0810211CF |
| HSV-1, Isolate #10 | 0810191CF | HSV-2, Isolate #11 | 0810212CF |
| HSV-1, Isolate #11 | 0810192CF | HSV-2, Isolate #12 | 0810213CF |
| HSV-1, Isolate #12 | 0810193CF | HSV-2, Isolate #13 | 0810214CF |
| HSV-1, Isolate #13 | 0810194CF | HSV-2, Isolate #14 | 0810215CF |
| HSV-1, Isolate #14 | 0810195CF | HSV-2, Isolate #15 | 0810216CF |
| HSV-1, Isolate #15 | 0810196CF | HSV-2, Isolate #16 | 0810217CF |
| HSV-1, Isolate #16 | 0810197CF | HSV-2, Isolate #17 | 0810218CF |
| HSV-1, Isolate #17 | 0810198CF | HSV-2, Isolate #18 | 0810219CF |
| HSV-1, Isolate #18 | 0810199CF | HSV-2, Isolate #19 | 0810220CF |
| HSV-1, Isolate #19 | 0810200CF | HSV-2, Isolate #20 | 0810221CF |
| HSV-1, Isolate #20 | 0810201CF | HSV-2, Isolate #21 | 0810222CF |
| HSV-1, Isolate #21 | 0810202CF | | |
# e. Analytical specificity:
# Cross Reactivity:
A panel of microorganisms that may be present in patient specimens was tested to determine whether these microorganisms interfered with the detection of HSV-1 or HSV-2 or were cross-reactive with the artus HSV-1/2 QS-RGQ MDx Kit. Organisms were tested at a target concentration of approximately $1 \times 10^{6}$ CFU/ml for bacteria (with the exception of Neisseria gonorrhea, which was tested at $3.5 \times 10^{5}$ CFU/mL due to the low titer of the stock culture) and fungi or $\geq 1 \times 10^{5}$ TCID $_{50}$ /ml for viruses separately in the presence of 2-3X LoD of each of three HSV strains: HSV-1 MacIntyre, HSV-1 Isolate #15, or HSV-2 MS. None of the potential interfering organisms cross-reacted or interfered with the detection of any of the HSV strains by the artus HSV-1/2 QS-RGQ MDx Kit.
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Organisms Tested in Cross Reactivity and Microbial Interference
| Organism | Source ID |
| --- | --- |
| Acinetobacter calcaceticus | ATCC 51432 |
| Acinetobacter lwoffii | ATCC 17925 |
| Adenovirus type 2 | ZMC 0810110CF |
| Bacteroides fragilis | ZMC 0601533 |
| Candida albicans | ATCC 10231 |
| Candida glabrata | ZMC Z007 |
| Candida guilliermondii | ZMC Z008 |
| Candida krusei | ZMC Z009 |
| Candida lusitaniae | ATCC 42720 |
| Candida parapsilosis | ZMC Z011 |
| Candida tropicalis | ZMC Z012 |
| Chlamydia trachomatis | ATCC VR-885 |
| Cytomegalovirus | ZMC 0810003CF |
| Enterobacter cloacae | ATCC 13047 |
| Enterovirus | ZMC 0810047CF |
| Epstein-Barr Virus | ZMC 0810008CF |
| Escherichia coli | ATCC 23571 |
| Fusobacterium nucleatum | ATCC 25586 |
| Gardnerella vaginalis | ATCC 14019 |
| Haemophilus ducreyi | ATCC 700724D-5 |
| Human Genomic DNA | Promega G3041 |
| Human Herpes Virus 6 | ZMC 0810003CF |
| Human Herpes Virus 7 | ZMC 0810071CF |
| Human papilloma virus 16 | ATCC 45113 |
| Human papilloma virus 18 | ATCC 45152 |
| Herpes Simplex Virus 1 (HSV-1), isolate 20 | 0810201CF |
| Herpes Simplex Virus 2 (HSV-2), isolate 20 | 0810221CF |
| Klebsiella pneumoniae | ATCC 13883 |
| Lactobacillus acidophilus | ATCC 4356 |
| Mobiluncus curtsii | ATCC 43063 |
| Mobiluncus mulieris | ATCC 35240 |
| Moraxella catarrhalis | ATCC 8176 |
| Mycoplasma hominis | ATCC 23114D |
| Neisseria gonorrhea | ATCC 9793 |
| Neisseria meningitides | ATCC 13077 |
| Prevotella melaninogenica | ATCC 25845 |
| Rubella Virus | ZMC 0810048CF |
| Simian Virus type 40 (SV40) | VRMC-2 |
{12}
| Organism | Source ID |
| --- | --- |
| Staphylococcus aureus (MRSA) | ATCC 43300 |
| Staphylococcus aureus (MSSA) | ATCC 29213 |
| Staphylococcus epidermidis (MRSE) | ATCC 51625 |
| Staphylococcus saprophyticus | ATCC 15305 |
| Streptococcus mitis | ZMC Clinical isolate |
| Streptococcus mutans | ZMC Z072 |
| Streptococcus pneumoniae | ZMC 19F-Z022 |
| Streptococcus pyogenes | ATCC 8669 |
| Streptococcus salivarius | ATCC 13419 |
| Toxoplasma gondii | ZMC 0810007CF |
| Treponema pallidum | ATCC 632912 |
| Trichomonas vaginalis | ATCC PRA-98D |
| Varicella-Zoster Virus (VZV) | ZMC 0810171CF |
# Target Carryover Study:
Absence of carryover between samples for the entire workflow was demonstrated by performing 5 runs with alternating high positive $(\geq 1.0 \times 10^{5} \mathrm{TCID}_{50} / \mathrm{mL})$ and negative samples. All samples were detected correctly, generating a carryover rate of $0.0\%$ .
# Interfering Substances:
A panel of 24 substances that may be present in oral/genital patient specimens was tested to determine whether these substances interfered with the performance of the artus HSV-1/2 QS-RGQ MDx Kit. Two strains of HSV: HSV-1 MacIntyre and HSV-2 MS, were diluted to approximately 2–3X LOD in M4RT viral transport medium and spiked with each potentially inhibitory substance. None of the substances showed an inhibitory effect on the detection of HSV-1 or HSV-2 by the artus HSV-1/2 QS-RGQ MDx Kit.
Potentially Interfering Substances Tested
| Substance | Potential Interferent/Active Ingredient | Concentration Of Substance Added To Reaction | Concentration Of Active Ingredient Added To Reaction |
| --- | --- | --- | --- |
| Whole blood with EDTA | Hemoglobin, lactoferin | 100% | 5% v/v |
| Buffy coat | White blood cells | 100% | 5% v/v |
| Acyclovir | Acycloguanosine | 5 mg/mL | 5 mg/mL |
| Albumin | Albumin | 5 mg/mL | 5 mg/mL |
| Casein | Casein | 5 mg/mL | 5 mg/mL |
{13}
| Substance | Potential Interferent/Active Ingredient | Concentration Of Substance Added To Reaction | Concentration Of Active Ingredient Added To Reaction |
| --- | --- | --- | --- |
| Female urine* | Urea | 100% | Not listed |
| Male urine* | Urea | 100% | Not listed |
| K-Y® Brand jelly | Glycerin, hydroxyethyl cellulose, chlorhexidine, gluconate, gluconolactone, methylparaben, sodium hydroxide | 100% | Not listed |
| Douche* | Octoxynol-9, citric acid, sodium benzoate, disodium EDTA | 100% | Not listed |
| Spermicide* | Nonoxynol-9 | 100% | 7% |
| Yeast·Gard®* | Candida albicans ×27 HPUS†, Candida parapsilosis ×27 HPUS, Pulsatilla ×27 HPUS | 100% | Not listed |
| Monistat® 1* | Miconazole nitrate | 100% | 2% v/v |
| Vagisil® Cream* | Benzocaine, resorcinol | 100% | Benzocaine 20% Resorcinol 3% |
| Monistat 3* | Miconazole nitrate | 100% | 2% v/v |
| Triconazole 1* | Triconazole | 100% | 6.5% v/v |
| Rite Aid Feminine Wash, Sensitive Skin* | Ammonium laureth sulfate, ammonium lauryl sulfate, decyl glucoside,cocamidopropyl betaine | 100% | Not listed |
| Clotrimazole-7 vaginal cream* | Clotrimazole | 100% | 1% v/v |
| Anti-itch cream* | Benzocaine | 100% | 5% v/v |
| Listerine® antiseptic mouthwash* | Eucalyptol, menthol, methyl salicylate, thymol | 100% | Eucalyptol 0.920% Menthol 0.042% Methyl salicylate 0.060% Thymol 0.064% |
| Abreva®* | Docosanol | 100% | 10% v/v |
| Carmex® lip balm* | Menthol, camphor, phenol | 100% | Menthol 0.7% Camphor 1.7% Phenol 0.4% |
| Releev® cold sore treatment* | Benzalkonium chloride | 100% | 0.13% v/v |
| Rubber® | Methyl salicylate | 100% | 0.02% mL |
| Spermicide® | Methyl salicylate | 100% | 0.02% mL |
{14}
| Substance | Potential Interferent/Active Ingredient | Concentration Of Substance Added To Reaction | Concentration Of Active Ingredient Added To Reaction |
| --- | --- | --- | --- |
| Lip Clear® lysine+* | Zinc oxide | 100% | 1.2% v/v |
| Toothpaste* | Stannous fluoride | 100% | 0.454% v/v |
*: Applied directly to sample by swab.
†: HPUS: Homeopathic Pharmacopeia of the United States.
f. Assay cut-off:
See limit of detection.
2. Comparison studies:
a. Method comparison with predicate device:
Not applicable, see clinical study
b. Matrix comparison:
Not Applicable
3. Clinical studies:
a. Clinical Sensitivity:
See other clinical supportive data
b. Clinical specificity:
See other clinical supportive data
c. Other clinical supportive data (when a. and b. are not applicable):
Prospective Study:
The performance of the artus HSV-1/2 QS-RGQ MDx Kit was evaluated at 3 testing sites in 2013-2014, using samples from 5 geographically diverse locations within the United States. A total of 662 male and female genital or oral lesion swabs (510 genital and 152 oral) prospectively collected from symptomatic patients was evaluated. Results from the artus HSV-1/2 QS-RGQ MDx Kit were compared to results obtained from the ELVIS® (Enzyme Linked Virus Inducible System) HSV ID and D3 Typing Test System (Diagnostic Hybrids, Athens, OH).
15
{15}
Ninety-six (96) prospective specimens identified as HSV-2 positive by ELVIS viral culture were removed from the initial 510 genital specimens for the calculation of the HSV-1 clinical performance. As a result, there were 566 samples (414 genital and 152 oral) used to determine clinical performance for HSV-1. The clinical performance for HSV-1 and HSV-2 mucocutaneous lesions was also assessed. Prospectively collected samples that were classified as mucocutaneous included oral, cervical, vaginal, rectal, and any sample taken from a vesicle or lesion. The performance information is presented in the following tables:
HSV-1 Results for Genital Samples (N=414)
| HSV-1 | ELVIS | | | |
| --- | --- | --- | --- | --- |
| | | POS | NEG | Total |
| artus HSV-1/2 QS-RGQ MDx Kit | POS | 69 | 21* | 90† |
| | NEG | 3** | 321 | 324 |
| | TOTAL | 72 | 342 | 414 |
| 95% CI | | | | |
| Sensitivity – 95.8% (69/72) | | 88.5% – 98.6% | | |
| Specificity – 93.9% (321/342) | | 90.8% – 96.0% | | |
| Positive Predictive Value – 76.7% | | 67.0% – 84.2% | | |
| Negative Predictive Value – 99.1% | | 97.3% – 99.7% | | |
| Prevalence – 17% | | 14% – 21% | | |
*15 of the 21 discordant specimens (artus HSV-1/2 QS-RGQ MDx Positive, ELVIS Negative) were positive for HSV-1 by alternative PCR followed by bi-directional sequencing.
**All 3 discordant specimens (artus HSV-1/2 QS-RGQ MDx Negative, ELVIS Positive) were negative for HSV-1 by alternative PCR followed by bi-directional sequencing.
† 5 samples were undetermined for ELVIS out of a total of 95 HSV-1 artus positive genital samples.
HSV-2 Results for Genital Samples (N=510)
| HSV-2 | ELVIS | | | |
| --- | --- | --- | --- | --- |
| | | POS | NEG | Total |
| artus HSV-1/2 QS-RGQ MDx Kit | POS | 93 | 37* | 130 |
| | NEG | 3** | 377 | 380 |
| | TOTAL | 96 | 414 | 510 |
| 95% CI | | | | |
| Sensitivity – 96.9% (93/96) | | 91.2% – 98.9% | | |
| Specificity – 91.1% (377/414) | | 87.9% – 93.5% | | |
| Positive Predictive Value – 71.5% | | 63.3% – 78.6% | | |
| Negative Predictive Value – 99.2% | | 97.7% – 99.7% | | |
| Prevalence – 19% | | 16% – 22% | | |
*27 of the 32 discordant specimens (artus HSV-1/2 QS-RGQ MDx Positive, ELVIS Negative) were positive for HSV-2 by alternative PCR followed by bi-directional sequencing. The remaining 5 samples were not available for discordant analysis.
**All 3 discordant specimens (artus HSV-1/2 QS-RGQ MDx Negative, ELVIS Positive) were negative for HSV-2 by alternative PCR followed by bi-directional sequencing.
{16}
HSV-1 Results for Oral Samples (N=152)
| HSV-1 | ELVIS | | | |
| --- | --- | --- | --- | --- |
| | | POS | NEG | Total |
| artus HSV-1/2 QS-RGQ MDx Kit | POS | 45 | 19* | 64 |
| | NEG | 3** | 85 | 88 |
| | TOTAL | 48 | 104 | 152 |
| 95% CI | | | | |
| Sensitivity – 93.8% (45/48) | | 83.2% – 97.9% | | |
| Specificity – 81.7% (85/104) | | 73.2% – 88.0% | | |
| Positive Predictive Value – 70.3% | | 58.2% – 80.1% | | |
| Negative Predictive Value – 96.6% | | 90.5% – 98.8% | | |
| Prevalence – 32% | | 25% – 39% | | |
*13 of the 19 discordant specimens (artus HSV-1/2 QS-RGQ MDx Positive, ELVIS Negative) were positive for HSV-1 by alternative PCR followed by bi-directional sequencing.
** 2 discordant specimens (artus HSV-1/2 QS-RGQ MDx Negative, ELVIS Positive) were negative for HSV-1 by alternative PCR followed by bi-directional sequencing. The remaining 1 specimen was unavailable for discordant analysis testing.
HSV-2 Results for Oral Samples (N=152)
| HSV-2 | ELVIS | | | |
| --- | --- | --- | --- | --- |
| | | POS | NEG | Total |
| artus HSV-1/2 QS-RGQ MDx Kit | POS | 0 | 0 | 0 |
| | NEG | 0 | 152 | 152 |
| | TOTAL | 0 | 152 | 152 |
| 95% CI | | | | |
| Sensitivity – N/A | | N/A | | |
| Specificity – 100% (152/152) | | 97.5% – 100% | | |
| Positive Predictive Value – N/A | | N/A | | |
| Negative Predictive Value – 100% | | 97.5% – 100% | | |
| Prevalence – 0% | | 0% – 2% | | |
HSV-1 Mucocutaneous Samples (N=281)
| HSV-1 | ELVIS | | | |
| --- | --- | --- | --- | --- |
| | | POS | NEG | Total |
| artus HSV-1/2 QS-RGQ MDx Kit | POS | 62 | 23 | 85 |
| | NEG | 4 | 192 | 196 |
| | TOTAL | 66 | 215 | 281 |
| 95% CI | | | | |
| Sensitivity – 93.9% (62/66) | | 85.4% – 97.6% | | |
| Specificity – 89.3% (192/215) | | 84.5% – 92.8% | | |
| Positive Predictive Value – 72.9% | | 62.7% –81.2% | | |
| Negative Predictive Value – 98.0% | | 94.9% – 99.2% | | |
| Prevalence – 23% | | 19% – 29% | | |
{17}
HSV-2 Mucocutaneous Samples (N=320)
| HSV-2 | ELVIS | | | |
| --- | --- | --- | --- | --- |
| | | POS | NEG | Total |
| artus HSV-1/2 QS-RGQ MDx Kit | POS | 37 | 16 | 53 |
| | NEG | 2 | 265 | 267 |
| | TOTAL | 39 | 281 | 320 |
| | | 95% CI | | |
| Sensitivity – 94.9% (37/39) | | 83.1% – 98.6% | | |
| Specificity – 94.3% (265/281) | | 91.0% – 96.5% | | |
| Positive Predictive Value – 69.8% | | 56.5% – 80.5% | | |
| Negative Predictive Value – 99.3% | | 97.3% – 99.8% | | |
| Prevalence – 12% | | 9% – 16% | | |
HSV-2 Oral Retrospective Sample Study and Contrived Sample Study:
A retrospective study was conducted using oral samples for HSV2 detection. A total of 38 oral retrospective specimens were tested with the artus HSV-1/2 QS-RGQ MDx assay and ELVIS HSV ID and D' Typing Test System. There were no HSV-2 positive specimens detected in 38 oral specimens.
A contrived specimen study was performed to provide additional performance data for detection of HSV-2 in oral samples. A panel of seventy (70) individual samples consisting of 15 HSV-1/2 negative oral samples, 10 HSV-1 positive oral samples, and 45 HSV-1/2 negative oral samples spiked with HSV-2 at a concentration from 3X LoD to 1000X LoD was tested with the artus HSV-1/2 QS-RGQ MDx Kit. The HSV-1 positive oral samples and HSV-1/2 negative oral samples obtained from the method comparison study were used to make the panel. All samples were randomized and blinded to the operator prior to testing. HSV-2 was detected in all 45 contrived samples at all concentrations tested, supporting the claim for detection of HSV-2 in oral samples by the artus HSV-1/2 QS-RGQ MDx Kit.
4. Clinical cut-off:
Not Applicable
5. Expected values/Reference range:
The observed expected values for HSV-1 and HSV-2 during a multi-center clinical trial were calculated for the artus HSV-1/2 QS-RGQ MDx kit. The expected values for the patients ages 18 and older are shown for genital lesion samples and for all ages for the oral samples. The observed prevalence rates for HSV-1 were estimated as 18.6% (91/489) for genital samples and 42.1% (64/152) for oral samples. The prevalence rates for HSV-2 were estimated as 26.2% (128/489) for genital samples and 0% (0/152) for oral samples. Gender and age distribution is provided in the following tables.
{18}
Age Distribution for Genital Specimens
| Age (Years) | Total # of specimens | HSV-1 Positive | HSV-2 Positive | Negative |
| --- | --- | --- | --- | --- |
| 18-20 | 48 | 11 | 13 | 24 |
| 21-30 | 167 | 32 | 40 | 95 |
| 31-40 | 111 | 18 | 31 | 62 |
| 41-50 | 69 | 12 | 15 | 42 |
| 51-60 | 48 | 10 | 15 | 23 |
| 61-70 | 31 | 4 | 12 | 15 |
| 71-80 | 11 | 4 | 2 | 5 |
| 81-90 | 3 | 0 | 0 | 3 |
| 91-97 | 1 | 0 | 0 | 1 |
| Total | 489 | 91/489 | 128/489 | 270/489 |
Age Distribution for Oral Specimens
| Age (Years) | Total # of specimens | HSV-1 Positive | HSV-2 Positive | Negative |
| --- | --- | --- | --- | --- |
| < 1 | 1 | 1 | 0 | 1 |
| 1-10 | 21 | 15 | 0 | 6 |
| 11-20 | 20 | 6 | 0 | 14 |
| 21-30 | 25 | 8 | 0 | 17 |
| 31-40 | 23 | 9 | 0 | 14 |
| 41-50 | 16 | 3 | 0 | 13 |
| 51-60 | 14 | 5 | 0 | 9 |
| 61-70 | 15 | 6 | 0 | 9 |
| 71-80 | 11 | 6 | 0 | 5 |
| 81-90 | 2 | 2 | 0 | 0 |
| 91-97 | 4 | 3 | 0 | 1 |
| Total | 152 | 64/152 | 0/152 | 88/152 |
Positive and Negative Predictive Value: Hypothetical positive and negative predictive values (PPV & NPV) for the artus HSV-1/2 QS-RGQ MDx kit are shown in the table below. These calculations are based on hypothetical prevalence and overall sensitivity and specificity per specimen type as determined in the clinical study.
For HSV-1, these calculations are based upon an overall sensitivity and specificity of 96% and 94%, respectively, for genital swabs and 94% and 82%, respectively, for oral swabs.
For HSV-2, these calculations are based upon an overall sensitivity and specificity of 97% and 91%, respectively, for genital swabs and 0.0% and 100%, respectively, for oral swabs.
PPV was calculated using: (Sensitivity x Prevalence) / (Sensitivity x Prevalence + [1 - Specificity] x [1 - Prevalence]).
NPV was calculated using: (Specificity x [1 - Prevalence]) / ([1 - Sensitivity] x Prevalence + Specificity x [1 - Prevalence]).
{19}
Positive and Negative Predictive Values (PPV & NPV) for the artus assay based on sample type
| Prevalence (%) | Genital Swabs | | | | Oral Swabs | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | HSV-1 | | HSV-2 | | HSV-1 | | HSV-2 | |
| | PPV (%) | NPV (%) | PPV (%) | NPV (%) | PPV (%) | NPV (%) | PPV (%) | NPV (%) |
| 2 | 24.6% | 99.9% | 18.0% | 99.9% | 9.6% | 99.9% | N/A | 98.0% |
| 5 | 45.7% | 99.8% | 36.2% | 99.8% | 21.6% | 99.6% | N/A | 95.0% |
| 10 | 64.0% | 99.5% | 54.5% | 99.6% | 36.7% | 99.2% | N/A | 90.0% |
| 20 | 80.0% | 98.9% | 72.9% | 99.2% | 56.6% | 98.2% | N/A | 80.0% |
| 30 | 87.3% | 98.2% | 82.2% | 98.6% | 69.1% | 97.0% | N/A | 70.0% |
| 40 | 91.4% | 97.2% | 87.8% | 97.8% | 77.7% | 95.3% | N/A | 60.0% |
| 50 | 94.1% | 95.9% | 91.5% | 96.8% | 83.9% | 93.2% | N/A | 50.0% |
N/A = Not Applicable
Mucocutaneous Lesions: A subset of samples from the clinical study was identified as mucocutaneous. Those classified as mucocutaneous included: oral, cervical, vaginal, rectal. The following table lists the specific locations for the mucocutaneous lesions that were reported as such in the study along with the total number of samples from each specific location and number of positives.
Mucocutaneous Lesion Sites
| Breakdown of Mucocutaneous Samples | | HSV-1 Culture Positive | HSV-1 artus Positive | HSV-1 Concordant Positive | HSV-2 Culture Positive | HSV-2 artus Positive | HSV-2 Concordant Positive |
| --- | --- | --- | --- | --- | --- | --- | --- |
| Location | Total | | | | | | |
| Bottom Lip | 1 | | | | | | |
| Cervical | 21 | 3 | 3 | 2 | 1 | 2 | 1 |
| Clitoral | 2 | | | | | | |
| Corners Of Lips | 1 | | | | | | |
| Penis foreskin | 1 | | | | | | |
| Genital | 8 | 2 | 3 | 2 | | 1 | |
| Groin Vesicles | 1 | | | | | | |
| In-Mouth | 1 | 1 | 1 | 1 | | | |
| Vaginal Introitus | 1 | | | | | | |
| L Nares | 1 | | | | | | |
| Labia | 47 | 3 | 5 | 3 | 9 | 12 | 9 |
| Labia Major | 1 | | | | 1 | 1 | 1 |
| Left Labia | 5 | | | | 1 | 1 | 1 |
| Left Outer Labia | 1 | | | | | | |
| Left Upper Palate | 1 | | | | | | |
| Left Vulvar | 1 | | | | | | |
{20}
| Breakdown of Mucocutaneous Samples | | HSV-1 Culture Positive | HSV-1 artus Positive | HSV-1 Concordant Positive | HSV-2 Culture Positive | HSV-2 artus Positive | HSV-2 Concordant Positive |
| --- | --- | --- | --- | --- | --- | --- | --- |
| Location | Total | | | | | | |
| Lip | 31 | 17 | 20 | 17 | | | |
| Mons Pubic, Clitoris | 1 | | | | | 1 | |
| Mouth | 35 | 10 | 14 | 10 | | | |
| Oral Blister | 3 | | 2 | | | | |
| Palate | 2 | 1 | | | | | |
| Penile Lesion | 2 | | | | | 1 | |
| Rectal | 3 | | | | 1 | 1 | 1 |
| Right Labia Papule | 2 | | | | 1 | 1 | 1 |
| Right Side Of Mouth | 1 | 1 | 1 | 1 | | | |
| Throat | 7 | 2 | 1 | 1 | | | |
| Tongue | 6 | 2 | 3 | 2 | | | |
| Tooth | 1 | 1 | 1 | 1 | | | |
| Ulcer In Mouth | 1 | 1 | 1 | 1 | | | |
| Upper Hard Palate | 1 | | 1 | | | | |
| Upper Lip | 4 | 2 | 3 | 2 | | | |
| Urethral | 8 | | 1 | | 2 | 2 | 2 |
| Urogenital | 4 | 1 | 1 | 1 | | | |
| Vagina | 65 | 11 | 11 | 10 | 17 | 20 | 15 |
| Vaginal Rectal | 9 | 1 | 2 | 1 | | 1 | |
| Vesicle | 6 | | 2 | | | 1 | |
| Vulva | 34 | 7 | 9 | 7 | 6 | 8 | 6 |
| Total | 320 | 66 | 85 | 62 | 39 | 53 | 37 |
# N. Proposed Labeling:
The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.
# O. Conclusion:
The submitted information in this premarket notification is complete and supports a substantial equivalence decision.
