SEEGENE ANYPLEX II HSV-1/2 ASSAY

K142156 · Seegene · OQO · Feb 13, 2015 · Microbiology

Device Facts

• Record ID: K142156
• Device Name: SEEGENE ANYPLEX II HSV-1/2 ASSAY
• Applicant: Seegene
• Product Code: OQO · Microbiology
• Decision Date: Feb 13, 2015
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3305
• Device Class: Class 2

Intended Use

The Anyplex™ II HSV-1/2 Assay is a real-time polymerase chain reaction (PCR)-based in vitro diagnostic test intended for the qualitative detection and differentiation of Herpes Simplex Virus Type-1 (HSV-1) and Herpes Simplex Virus Type-2 (HSV-2) DNA from female skin lesions from anogenital sites. The test is intended for use as an aid in the diagnosis of anogenital HSV infection in symptomatic patients. Warning: Anyplex™ II HSV-1/2 Assay is not FDA cleared for use with cerebrospinal fluid (CSF). The assay is not intended to be used for prenatal screening.

Device Story

The Anyplex™ II HSV-1/2 Assay is an in vitro diagnostic test for qualitative detection and differentiation of HSV-1 and HSV-2 DNA. Input: female anogenital skin lesion specimens. Process: manual DNA extraction using QIAGEN QIAamp DNA Mini Kit; real-time PCR amplification on Cepheid SmartCycler® II Dx instrument; detection via fluorescently-labeled duplex Catcher probes. Output: qualitative detection of HSV-1 and HSV-2. Used in clinical laboratories by trained personnel. Results aid clinicians in diagnosing anogenital HSV infection in symptomatic patients. Includes positive, negative, and blank negative controls to ensure run validity and correct sample preparation.

Clinical Evidence

Clinical performance evaluated at three US sites using 656 prospective female anogenital specimens compared to ELVIS® HSV ID/Typing Test System. HSV-1 sensitivity: 98.9% (95% CI: 94.1-99.8%), specificity: 93.7% (95% CI: 91.0-95.6%). HSV-2 sensitivity: 97.2% (95% CI: 92.0-99.0%), specificity: 93.6% (95% CI: 91.3-95.4%). Discordant analysis performed via bidirectional sequencing.

Technological Characteristics

Real-time PCR-based assay. Uses fluorescently-labeled duplex Catcher probes for detection. Manual DNA extraction via QIAamp DNA Mini Kit. Instrumentation: Cepheid SmartCycler II Dx. Controls: HSV-1/2 plasmid positive control, internal control (IC) for extraction/process monitoring, and negative controls. Qualitative detection and differentiation of HSV-1 and HSV-2 DNA.

Indications for Use

Indicated for symptomatic female patients with anogenital skin lesions to qualitatively detect and differentiate HSV-1 and HSV-2 DNA. Not for use with 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

• IMDx HSV-1/2 for Abbott m2000 assay (K140198)

Reference Devices

• ELVIS® HSV ID/Typing Test System (K971662)

Submission Summary (Full Text)

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Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features the department's name encircling a symbol. The symbol consists of a stylized caduceus, with three human profiles facing right, representing health and well-being. The text reads "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA".

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

August 17, 2015

SEEGENE C/O FRAN WHITE, REGULATORY CONSULTANT MDC ASSOCIATES, LLC 180 CABOT STREET BEVERLY MA 01915

Re: K142156

Trade/Device Name: Anyplex™ II HSV-1/2 Assay Regulation Number: 21 CFR 866.3305 Regulation Name: Herpes Simplex Virus Nucleic Acid Amplification Assay Regulatory Class: II Product Code: OQO Dated: February 10, 2015 Received: February 11, 2015

Dear Dr. White:

This letter corrects our substantially equivalent letter of February 13, 2015.

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, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of

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medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) 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 you desire specific advice for your device on our labeling regulation (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number [phone] or [phone] or at its Internet address

http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR 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 [phone] or [phone] or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

# 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

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# Indications for Use

510(k) Number (if known) K142156

Device Name AnyplexTM II HSV-1/2 Assay

#### Indications for Use (Describe)

The AnyplexTM II HSV-1/2 Assay is a real-time polymerase chain reaction (PCR)-based in vitro diagnostic test intended for the qualitative detection and differentiation of Herpes Simplex Virus Type-1 (HSV-1) and Herpes Simplex Virus Type-2 (HSV-2) DNA from female skin lesions from anogenital sites. The test is intended for use as and in the diagnosis of anogenital HSV infection in symptomatic patients.

WARNING: The AnyplexTM II HSV-1/2 Assay is not indicated for use with cerebrospinal fluid (CSF). The assay is not intended to be used for prenatal screening.

| Type of Use (Select one or both, as applicable) | |
|-------------------------------------------------|--|
| | |

X Prescription Use (Part 21 CFR 801 Subpart D)

| | Over-The-Counter Use (21 CFR 801 Subpart C)

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## 510(k) Summary

| Date of Summary: | February 9, 2015 |
|----------------------------|---------------------------------------------------------------------------------------------------|
| Product Name | Anyplex™ II HSV-1/2 Assay |
| Sponsor | Seegene<br>Taewon Building, 91 Ogeum-ro Songpa-Gu<br>Seoul, South Korea, 138-050 |
| Correspondent | MDC Associates, LLC<br>Fran White, Regulatory Consultant<br>180 Cabot Street<br>Beverly, MA 01915 |
| Device Identification | |
| Trade or Proprietary Name: | Anyplex™ II HSV-1/2 Assay |
| Common or Usual Name: | HSV-1/2 Assay |
| Product Code: | OQO |
| Regulation Section: | 21 CFR 866.3305 |
| Product Classification: | Class II |

### Intended Use

The Anyplex 110 II HSV-1/2 Assay is a real-time polymerase chain reaction (PCR)-based in vitro diagnostic test intended for the qualitative detection and differentiation of Herpes Simplex Virus Type-1 (HSV-1) and Herpes Simplex Virus Type-2 (HSV-2) DNA from female skin lesions from anogenital sites. The test is intended for use as an aid in the diagnosis of anogenital HSV infection in symptomatic patients.

WARNING: The Anyplex™ II HSV-1/2 Assay is not indicated for use with cerebrospinal fluid (CSF). The assay is not intended to be used for prenatal screening.

### Device Description

The Anyplex™ II HSV-1/2 Assay uses PCR to generate amplified product from HSV-1 and HSV-2 present in clinical specimens. The presence of HSV-1 and/or HSV-2 target DNA is indicated by the fluorescent signal generated through the use of fluorescently-labeled oligonucleotide probes (duplex Catcher) on the Cepheid SmartCycler® II Dx instrument. The

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probes do not generate a signal unless they are specifically bound to the amplified product. A preparation of HSV-1 and HSV-2 plasmids is included as the positive control in the Anyplex™ II HSV-1/2 Assay. Run as a separate control, the positive control serves to demonstrate that the HSV-1/2 PCR reagents are functional, and discriminate the validity of the run. In addition, the positive control functions as a process control, demonstrating that sample preparation has proceeded correctly during the run. An internal control (IC) is also included in the assay kit. The IC is added to each sample specimen during sample preparation, and is also used to create a Blank Negative Control by adding a set amount to viral transport media to serve as an extraction control. In addition, the RNase-free water is used to create the Negative Control by adding a set volume to the prepared master mix. Users are instructed to include all three controls, Positive, Negative, and Blank Negative Control with each test run.

### Substantial Equivalency

The Anyplex™ II HSV-1/2 Assay is substantially equivalent to the IMDx HSV-1/2 for Abbott m2000 assay. Table 1 compares the characteristics of the Anyplex™ II HSV-1/2 Assay (New Device) and the IMDx HSV-1/2 for Abbott m2000 assay (Predicate Device).

| Similarities | | |
|----------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Characteristic | IMDx HSV-1/2 for<br>Abbott m2000 Assay<br>(Predicate Device) | Anyplex™ II HSV-1/2 Assay<br>(New Device) |
| 510(k) | K140198 | K142156 |
| Regulation | 21 CFR 866.3305 | 21 CFR 866.3305 |
| Product Code | OQO | OQO |
| Device Class | Class II | Class II |
| Intended use | The IMDx HSV-1/2 for Abbott m2000 assay is an <i>in vitro</i> diagnostic test for the direct, qualitative detection and differentiation of herpes simplex viruses type 1 (HSV-1) and type 2 (HSV-2) DNA from male and female skin lesions from anogenital or oral sites. The test is intended for use as an aid in the diagnosis of HSV infection in symptomatic patients. The assay is intended to be run on the Abbott m2000 instrument system.<br><br><b>Warning:</b> The IMDx HSV-1/2 for Abbott m2000 assay is not FDA cleared for use with cerebral spinal | The Anyplex™ II HSV-1/2 Assay is a real-time polymerase chain reaction (PCR)-based <i>in vitro</i> diagnostic test intended for the qualitative detection and differentiation of Herpes Simplex Virus Type-1 (HSV1) and Herpes Simplex Virus Type-2 (HSV2) DNA from female skin lesions from anogenital sites. The test is intended for use as an aid in the diagnosis of anogenital HSV infection in symptomatic patients.<br><br><b>WARNING:</b> The Anyplex™ II HSV-1/2 Assay is not indicated for use with cerebrospinal fluid (CSF). The |

### Table 1: Substantial Equivalence

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| Similarities | | | | | | |
|----------------------|------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------|--|--|--|--|
| Characteristic | IMDx HSV-1/2 for<br>Abbott m2000 Assay<br>(Predicate Device) | Anyplex™ II HSV-1/2 Assay<br>(New Device) | | | | |
| | fluid (CSF) or for pre-natal screening. | assay is not intended to be used for<br>prenatal screening. | | | | |
| | | | | | | |
| Test Principle | Real-time PCR DNA amplification | Real-time PCR DNA amplification | | | | |
| Assay Results | Qualitative detection and<br>differentiation of HSV-1 and HSV-2 | Qualitative detection and<br>differentiation of HSV-1 and HSV-2 | | | | |
| Differences | | | | | | |
| Characteristic | IMDx HSV-1/2 for<br>Abbott m2000 Assay<br>(Predicate Device) | Anyplex™ II HSV-1/2 Assay<br>(New Device) | | | | |
| Instrumentation | Sample extraction and real-time PCR<br>amplification/detection using the<br>Abbott m2000 system. | Real-time PCR<br>amplification/detection using the<br>Cepheid SmartCycler II DX system. | | | | |
| Extraction<br>Method | Automated on Abbott m2000 system | Manual extraction using the QIAGEN<br>QIAamp® DNA Mini Kit | | | | |
| Detection<br>Method | Double-labeled (fluorophore and<br>quencher) hydrolysis probes.<br>Measures increase in assay<br>fluorescence with each PCR cycle. | Double-labeled (fluorophore and<br>quencher) duplex Catcher. Measures<br>increase in assay fluorescence with<br>each PCR cycle. | | | | |
| Sample type | Male and female skin lesions from<br>anogenital or oral sites | Female skin lesions from anogenital<br>sites only | | | | |

### Performance Characteristics

## Analytical Performance

# Precision/Repeatability:

A precision study was conducted in-house by testing 5 different panels consisting of HSV-1/2 Negative, HSV-1 Low Positive (1X LoD), HSV-1 High Positive (10X LoD), HSV-2 Low Positive (1X LoD) and HSV-2 High Positive (10X LoD). All panels were tested twice per day for twenty days by one operator. Samples were tested in triplicates for each run (for a total of 600 data points for the 40 runs).

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| Panel Member | Level | Agreement<br>with expected<br>results | 95%<br>Confidence<br>Interval | Avg. Ct | SD Ct | %CV Ct |
|------------------------|---------|---------------------------------------|-------------------------------|---------|-------|--------|
| HSV-1 Low<br>Positive | 1X LoD | 100.00%<br>(120/120) | 96.90% -<br>100.00% | 42.93 | 0.75 | 1.74% |
| HSV-1 High<br>Positive | 10X LoD | 100.00%<br>(120/120) | 96.90% -<br>100.00% | 40.32 | 0.71 | 1.76% |
| HSV-2 Low<br>Positive | 1X LoD | 100.00%<br>(120/120) | 96.90% -<br>100.00% | 41.31 | 1.06 | 2.56% |
| HSV-2 High<br>Positive | 10X LoD | 100.00%<br>(120/120) | 96.90% -<br>100.00% | 38.19 | 0.51 | 1.34% |
| Negative | N/A | 100.00%<br>(120/120) | 96.90% -<br>100.00% | N/A | N/A | N/A |

Table 2: Precision Study Average Ct Values

Mean: Average Ct of positive results only.

### Precision/Reproducibility Results

Reproducibility studies were performed at three sites (one internal and two external clinical sites) using 3 lots of the reagent kits. Each test site tested one lot of the reagent kit. The test panel of seven members was prepared blinded and randomized and tested for five (5) days nonconsecutive days by two operators with each operator running the panel, in triplicate, once a day. Variables in the reproducibility study included between run-to-run, operator-to-operator, site-tosite and lot-to-lot.

Table 3 below shows the Percent (%) Agreement and Average of Ct for each site as well as the Total (%) agreement and Average of Ct for all 3 sites combined.

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# Table 3: Reproducibility Study Summary

| | | Site 1 | | Site 2 | | Site 3 | | All 3 sites Combined | |
|----------------|---------------|-----------------------------------------------------|------------------|--------------------------------------------------|------------------|-----------------------------------------------------|------------------|-----------------------------------------------------|------------------|
| Panel Member | Concentration | % Agreement<br>Agreement<br>with expected<br>result | Avg. Ct<br>(%CV) | % Agreement<br>Agreement with<br>expected result | Avg. Ct<br>(%CV) | % Agreement<br>Agreement<br>with expected<br>result | Avg. Ct<br>(%CV) | % Agreement<br>Agreement<br>with expected<br>result | Avg. Ct<br>(%CV) |
| HSV-1 High- | 3X LoD | 100.0%<br>30/30 | 36.4<br>(2.0%) | 100.0%<br>30/30 | 35.9<br>(2.6%) | 100.0%<br>30/30 | 36.1<br>(2.9%) | 100.0%<br>90/90 | 36.1<br>(2.6%) |
| Positive* | | | | | | | | | |
| HSV-1 Low- | 1X LoD | 100.0%<br>30/30 | 39.9<br>(2.2%) | 90.0%<br>27/30 | 39.8<br>(3.3%) | 100.0%<br>30/30 | 39.2<br>(1.7%) | 96.67%<br>87/90 | 39.6<br>(2.6%) |
| Positive* | | | | | | | | | |
| HSV-1 High- | <1X LoD | 6.67%<br>2/30 | 42.9<br>(3.2%) | 43.33%<br>13/30 | 42.6<br>(3.9%) | 3.33%<br>1/30 | 42.4<br>(2.4%) | 17.78%<br>16/90 | 42.6<br>(3.1%) |
| Negative** | | | | | | | | | |
| HSV-2 High- | 3X LoD | 100.0%<br>30/30 | 36.5<br>(1.6%) | 100.0%<br>30/30 | 35.7<br>(2.2%) | 100.0%<br>30/30 | 36.4<br>(1.7%) | 100.0%<br>90/90 | 36.2<br>(2.0%) |
| Positive* | | | | | | | | | |
| HSV-2 Low- | 1X LoD | 100.0%<br>30/30 | 39.8<br>(2.3%) | 93.3%<br>28/30 | 39.6<br>(2.4%) | 100.0<br>30/30 | 39.9<br>(2.4%) | 97.8%<br>88/90 | 39.8<br>(2.3%) |
| Positive* | | | | | | | | | |
| HSV-2 High- | <1X LoD | 30.0%<br>9/30 | 42.3<br>(2.8%) | 46.67%<br>14/30 | 42.1<br>(4.0%) | 16.67%<br>5/30 | 42.2<br>(2.5%) | 31.1%<br>28/90 | 42.2<br>(3.0%) |
| Negative** | | | | | | | | | |
| HSV Negative** | N/A | 100.0%<br>60/60 | 0.0 | 98.33%<br>59/60 | 42.7 | 100.0%<br>60/60 | 0.0 | 99.4%<br>179/180 | 42.7 |

*Expected result is positive. %CV calculated from results with non-zero Ct values.

** Expected result is negative. %CV calculated from results with non-zero Ct values.

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## Analytical Sensitivity (Limit of Detection)

The LoD is determined as the HSV-1/2 titer (TCIDs(mL) detected with a positivity rate of 95% or greater. The LoD of the Anyplex™ II HSV-1/2 Assay was determined for two strains of HSV-1 and two strains of HSV-2. The results, representative of the analytical sensitivity of the AnyplexTM II HSV-1/2 Assay, are summarized in Table 4.

| Strain | Limit of Detection (95% CI) |
|-----------------|-----------------------------|
| HSV-1 MacIntyre | 3.75X102 |
| HSV-1 HF | 1.88X102 |
| HSV-2 MS | 3.75x101 |
| HSV-2 G | 3.75x101 |

## Table 4: Limit of Detection

### Cross-Reactivity and Microbial Interference

A panel of 50 organisms was tested for cross-reactivity and interference with the Anyplex™ II HSV-1/2 Assay. Intermediate stocks of bacteria, yeast, viruses, and organism genomic DNA were prepared from quantitated stocks and then diluted to their final test concentration. All samples were prepared by diluting organisms or DNA into M4 viral transport medium. No strains tested were positive for HSV-1 or HSV-2 using the Anyplex™ II HSV-1/2 Assay.

### Table 5: Cross-Reactivity and Microbial Interference Panel

| Organism | Organism |
|------------------------------------|------------------------------------|
| Atopobium vaginae | Gardnerella vaginalis |
| Bacteroides fragilis | Human Herpes 6B virus (Z29 strain) |
| Candida albicans | Human Herpes 7 virus (SB strain) |
| Candida glabrata Z007 | Human Papilloma virus-16 (Caski) |
| Candida guilliemondii Z008 | Human Papilloma virus-18 (Hela) |
| Candida krusei Z009 | Klebsiella pneumoniae Z026 |
| Candida lusitaniae Z010 | Lactobacillus acidophlus |
| Candida parapsilosis Z011 | Lactobacillus crispatus |
| Candida tropicalis Z012 | Lactobacillus gasseri |
| Chlamydia trachomatis (D-UW3) | Lactobacillus jensenii |
| Chlamydia trachomatis (serotype E) | Moraxella catarrhalis Ne 11 |
| Chlamydia trachomatis (serotype F) | Mycoplasma hominis |
| Chlamydia trachomatis (serotype G) | Neisseria gonorrhoeae |
| Chlamydia trachomatis (serotype H) | Rubella virus |
| Chlamydia trachomatis (serotype I) | Serratia marcescens |
| Chlamydia trachomatis (serotype J) | Staphylococcus saprophyticus |
| Chlamydia trachomatis (serotype K) | Streptococcus mitis |
| Cytomegalovirus (AD169) | Streptococcus mutans Z072 |

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| Organism | Organism |
|-----------------------------------|----------------------------------|
| Enterococcus casseliflavus | Streptococcus oralis |
| Enterococcus faecalis | Streptococcus pneumoniae 19F |
| Enterococcus faecium | Streptococcus pyogenes Rosenbach |
| Enterococcus gallinarum | Toxoplasma gondii |
| Enterovirus (Type 71) | Trichomonas vaginalis |
| Epstein-Barr virus (B95-8 strain) | Ureaplasma urealyticum |
| Escherichia coli | Varicella Zoster virus |

### Interfering Substances

The interference study was conducted with the Anyplex™ II HSV-1/2 assay using a panel of twenty two (22) interfering substances that could be present in the female anogenital swab lesion specimens and interfere with the performance of the Anyplex " II HSV-1/2 assay.

The interfering substances were tested at concentrations at or above physiological levels or typical usage levels with HSV strains (HSV-1 MacIntyre and HSV-2 MS) at 3X LoD. None of the 22 substances showed detectable effect of interference, resulting in all samples being positive for HSV1 or HSV2.

### Clinical Performance Characteristics

The performance of the Anyplex™ II HSV-1/2 Assay was evaluated at three geographicallydiverse locations within the United States from 2013-2014. A total of 656 valid specimens was included in the final data set and analyzed for product performance as compared to results obtained from the ELVIS® (Enzyme Linked Virus Inducible System) HSV ID and D Typing Test System (Diagnostic Hybrids, Athens, OH). The reference ELVIS viral culture method used in this study is unable to detect co-infected specimens and cannot identify HSV-1 if HSV-2 is identified first. Consequently, if a specimen was positive for HSV-2, it was removed from the calculation of the HSV-1 clinical performance.

Assay performance for anogenital specimens is shown below for the 656 prospective specimens included in the study.

| HSV-1 Performance | | Reference Method | | |
|-------------------------------|-------|----------------------|-----|-------|
| | | POS | NEG | Total |
| Anyplex™ II HSV-<br>1/2 Assay | POS | 91 | 29a | 120 |
| | NEG | 1b | 429 | 430 |
| | Total | 92 | 458 | 550 |
| Sensitivity; 95% CI | | 98.9% (91/92); | | |
| | | 95% CI [94.1%-99.8%] | | |
| Specificity; 95% CI | | 93.7% (429/458); | | |
| | | 95% CI [91.0%-95.6%] | | |

### Table 6: HSV-1 Anogenital Results

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ª Discordant analysis (bidirectional sequencing) was performed on all 29 samples identified as HSV-1 positive by the Anyplex™ II HSV-1/2 Assay. HSV-1 was detected in 18 of the 29 samples. The remaining 11 specimens remained discordant (HSV-1 was not detected).

b Discordant analysis (bidirectional sequencing) was performed on the one sample identified as negative by the Anyplex™ II HSV-1/2 Assay. HSV-1 was detected in this sample.

| HSV-2 Performance | | Reference Method | | |
|---------------------------------------------|-------|------------------------------------------|-----|-------|
| | | POS | NEG | Total |
| Anyplex TM II HSV-<br>1/2 Assay | POS | 103 | 35c | 138 |
| | NEG | 3d | 515 | 518 |
| | Total | 106 | 550 | 656 |
| Sensitivity; 95% CI | | 97.2% (103/106);<br>95% CI [92.0%-99.0%] | | |
| Specificity; 95% CI | | 93.6% (515/550);<br>95% CI [91.3%-95.4%] | | |

#### Table 7: HSV-2 Anogenital Results

^ Discordant analysis (bidirectional sequencing) was performed on all 35 discordant specimens identified as HSV-2 positive by the Anyplex™ II HSV-1/2 Assay. HSV-2 was detected in 21 of the 35 specimens. The remaining 14 remained discordant (HSV-2 was not detected).

d Discordant analysis (bidirectional sequencing) was conducted for the three specimens identified as HSV-2 negative by the Anyplex™ II HSV-1/2 Assay. HSV-2 was not detected in any specimen.

#### Conclusions

The submitted information in this premarket notification is complete and supports a substantial equvalence decision.