ISOAMP HSV ASSAY

K111951 · Biohelix Corporation · OQO · Sep 27, 2011 · Microbiology

Device Facts

• Record ID: K111951
• Device Name: ISOAMP HSV ASSAY
• Applicant: Biohelix Corporation
• Product Code: OQO · Microbiology
• Decision Date: Sep 27, 2011
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3305
• Device Class: Class 2
• Attributes: Pediatric

Intended Use

The IsoAmp® HSV Assay is an in vitro diagnostic test for the direct, qualitative detection of the Herpes Simplex Virus (HSV-1 & HSV-2) DNA in male and female genital and oral lesions. The test is intended for use as an aid in diagnosis of HSV infection in symptomatic patients. Warning: The IsoAmp® HSV Assay is not FDA cleared for use with cerebrospinal fluid (CSF). The assay does not provide specific typing information to differentiate HSV-1 and HSV-2. The assay is not intended to be used for prenatal screening.

Device Story

IsoAmp® HSV Assay is an in vitro diagnostic test for qualitative detection of HSV-1 and HSV-2 DNA in genital and oral lesions. Process: 1) specimen dilution in viral transport medium; 2) isothermal Helicase-Dependent Amplification (HDA) of HSV glycoprotein B (gB) gene at 64°C using biotinylated primers; 3) detection via lateral-flow strip in a self-contained disposable cassette. Cassette contains anti-FITC and anti-DIG antibodies for visual colorimetric readout of Test (T) and Control (C) lines. Used in clinical settings; operated by laboratory personnel. Provides qualitative results (positive/negative/invalid) to aid clinicians in diagnosing HSV infection. Benefits include rapid, isothermal detection without complex thermal cycling equipment.

Clinical Evidence

Clinical performance evaluated at 5 US sites (2010-2011) using 994 samples (962 prospective, 32 retrospective). Compared against ELVIS® HSV ID/Typing Test System. For genital samples (n=803): sensitivity 97.1% (94.3-98.5% CI), specificity 93.4% (91.0-95.2% CI). For oral samples (n=159): sensitivity 93.8% (83.2-97.9% CI), specificity 87.4% (79.9-92.3% CI).

Technological Characteristics

Isothermal Helicase-Dependent Amplification (HDA) technology. Employs biotinylated primers and lateral-flow DNA detection strips coated with anti-FITC and anti-DIG antibodies. Self-contained Type II BESt™ disposable cassette. Qualitative colorimetric readout. No software or electronic components.

Indications for Use

Indicated for symptomatic male and female patients with genital or oral lesions suspected of HSV infection. Not for use with CSF, prenatal screening, or typing of HSV-1 vs HSV-2.

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)

Reference Devices

• ELVIS® HSV ID/Typing Test System

Submission Summary (Full Text)

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તાવિકા

#### 1.0 Submitter

Date of Summary: September 24, 2011 SEP 2 7 2011

IsoAmp® HSV Assay Product Name: Sponsor: BioHelix Corporation 500 Cummings Center Suite 5550 Beverly, MA 01915

Correspondent : MDC Associates, LLC Fran White, Regulatory Consultant 180 Cabot Street Beverly, MA 01915

#### 2.0 Device Identification

Trade or Proprietary Name: IsoAmp® HSV Assay Herpes simplex virus Assay Common or Usual Name: Product Code: oqo Regulation Section: 21 CFR 866.3305 Product Classification: Class II

#### 3.0 Substantial Equivalency

IsoAmp® HSV Assay is substantially equivalent to the EraGen Biosciences MultiCode®-RTx Herpes Simplex Virus 1 & 2 Kit (K100336). The table below identifies the characteristics of BioHelix Corporation's IsoAmp® HSV Assay (K111951) and the EraGen Biosciences MultiCode®-RTx Herpes Simplex Virus 1 & 2 Kit (Predicate Device).

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| Features | | BioHelix Corporation<br>IsoAmp® HSV Assay<br>(New Device) | SIMILARITIES | | EraGen Bioscience<br>Multicode® RTx Herpes Simplex Virus 1 & 2 Kit<br>(Predicate Device) |
|---------------------------------|-------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------|--|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| | | K111951 | | | K100336 |
| Intended use | | The IsoAmp® HSV Assay is an in vitro diagnostic test for<br>the direct, qualitative detection of herpes simplex virus<br>(HSV-1 & HSV-2) DNA in male and female genital and oral<br>lesions. The test is intended for use as an aid in diagnosis<br>of HSV infection in symptomatic patients.<br><br>Warning: The IsoAmp® HSV Assay is not FDA cleared for<br>use with cerebrospinal fluid (CSF). The assay does not<br>provide specific typing information to differentiate HSV-<br>1 and HSV-2. The assay is not intended to be used for<br>prenatal screening. | | | The MultiCode®-RTx HSV-1&2 Kit is a polymerase chain<br>reaction (PCR) -based qualitative in vitro diagnostic test for<br>the detection and typing of herpes simplex virus (HSV-1&2<br>DNA in vaginal lesions. It is indicated for use in the<br>detection and typing of HSV-1 or HSV-2 in vaginal lesion<br>swab specimens from symptomatic female patients as an<br>aid in the diagnosis of genital herpes infection.<br><br>Warning: The device is no FDA cleared for the use with<br>cerebral spinal fluid (CSF) or any lesions other than<br>vaginal. This assay is not intended to be used for male<br>penile specimens, for prenatal screening, or for females<br>under the age of 18 years. |
| Assay Results | | Qualitative | | | Qualitative |
| Analysis Software Provided | | No | | | Yes |
| Printed Results Report Provided | | No | | | No |
| Detection of HSV-1 and HSV-2 | | Yes | | | Yes |
| | DIFFERENCES | | | | |
| Methodology | | HDA (Helicase-Dependent Amplification) | | | Real-Time PCR |
| Typing of HSV-1 and HSV-2 | | No | | | Yes |
| Packaging | | The product is supplied as two (2) separate labeled<br>boxes. | | | The product is supplied in labeled, sterile tubes. The<br>outer container is a labeled box. |
| Kit Reagent Storage Conditions | | AKRC: ≤-15°C<br>NKC: 15-30°C | | | -15°C to -30°C |

Comparison of New Device with Predicate Device

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# 4.0 Product Description

The IsoAmp® HSV Assay consists of three major steps: 1) specimen preparation: 2) isothermal Helicase-Dependent Amplification (HDA) of the HSV glycoprotein B (gB) gene using biotinylated primers; and 3) detection of the amplified DNA by a target-specific hybridization probe via a colorimetric reaction on a lateral-flow strip which is embedded in a self-contained disposable cassette to prevent amplicon contamination.

Specimen preparation includes a simple dilution step in which specimens in viral transport medium are diluted 40-fold in dilution buffer. The diluted samples are mixed with HDA reagents. Incubation at 64°C results in the release of the HSV DNA and subsequent isothermal amplification of the target sequence. A competitive internal control (IC) is included in the Amplification Reagents to monitor inhibitory substances in negative samples, reagent failure or device failure.

After incubation for one hour, the amplified DNA is detected by two detection probes, one labeled with fluorescein isothiocyanate (FITC) for hybridizing to the HSV target and the other labeled with digoxigenin (DIG) for binding to the IC target. The hybrid of FITC-labeled probe and HSV amplicon is captured at the Test Line (T-Line) on the lateral-flow strip by anti-FITC antibodies, while the DIG-labeled IC amplicon is captured at the Control Line (C-Line) on the strip by anti-DIG antibodies. The biotin label in each amplicon captures the streptavidinconjugated color particles for visualization and the test result is shown as colored lines that are visually read.

The self-contained Type II BESt™ cassettes contain lateral-flow DNA detection strips coated with anti-FITC antibodies and anti-DIG antibodies that serve as T line and C line respectively in the assay. A positive result (detection of HSV DNA) is reported when the T line is visible through the detection window of the cassette. A negative result (no detection of HSV DNA) is reported when only the C line is displayed. The assay result is regarded as invalid when both the T line and C line are not present and the assay should be repeated.

#### 5.0 Indications for Use and Intended Use

The IsoAmp® HSV Assay is an in vitro diagnostic test for the direct, qualitative detection of herpes simplex virus (HSV-1 & HSV-2) DNA in male and female genital and oral lesions. The test is intended for use as an aid in diagnosis of HSV infection in symptomatic patients.

Warning: The IsoAmp® HSV Assay is not FDA cleared for use with cerebrospinal fluid (CSF). The assay does not provide specific typing information to differentiate HSV-1 and HSV-2. The assay is not intended to be used for prenatal screening.

# 6.0 Analytical Performance

l.

:

- Precision/Reproducibility:
The Precision/Reproducibility of the IsoAmp® HSV Assay was evaluated at three (3) test sites. A panel of seven (7) members was prepared containing one negative control sample (HSV negative pooled swab specimens) and six simulated HSV-1 and HSV-2 samples that included High Negative (below the assay limit of detection), Low Positive (near the assay limit of detection) and Moderate Positive (three times the assay limit of detection) samples. The panel (which included one replicate of each panel member), along with external HSV-1 and HSV-2 positive and negative controls, was tested at each site for five (5) days

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by two operators with each operator running the panel two times a day using a single lot of the IsoAmp® HSV Assay. One (1) site tested the panel using three (3) lots. Results of the Precision/Reproducibility study for the IsoAmp® HSV at three sites are presented in the table below.

| Overall Reproducibility Study | | | | | |
|------------------------------------|-------------------|-------------------|-------------------|---------------------------|-------------------------|
| Category | Site #1* | Site #2 | Site #3 | Overall Percent Agreement | 95% Confidence Interval |
| | Percent Agreement | Percent Agreement | Percent Agreement | | |
| HSV-1 High Negative | 13/60<br>22% | 13/20<br>65% | 6/20<br>30% | 32/100<br>32% | 24% - 42% |
| HSV-1 Low Positive | 60/60<br>100% | 19/20<br>95% | 20/20<br>100% | 99/100<br>99% | 94% - 100% |
| HSV-1 Moderate Positive | 60/60<br>100% | 20/20<br>100% | 20/20<br>100% | 100/100<br>100% | 96% - 100% |
| HSV-2 High Negative | 19/60<br>32% | 7/20<br>35% | 6/20<br>30% | 32/100<br>32% | 24% - 42% |
| HSV-2 Low Positive | 60/60<br>100% | 18/20<br>90% | 18/20<br>89% | 96/100<br>96% | 90% - 98% |
| HSV-2 Moderate Positive | 60/60<br>100% | 20/20<br>100% | 20/20<br>100% | 100/100<br>100% | 96% - 100% |
| Negative1 | 60/60<br>100% | 20/20<br>100% | 19/20<br>95% | 99/100<br>99% | 96% - 100% |
| HSV-1 Positive Control | 60/60<br>100% | 20/20<br>100% | 20/20<br>100% | 100/100<br>100% | 96% - 100% |
| HSV-2 Positive Control | 60/60<br>100% | 20/20<br>100% | 20/20<br>100% | 100/100<br>100% | 96% - 100% |
| Assay Negative Control2 | 60/60<br>100% | 20/20<br>100% | 20/20<br>100% | 100/100<br>100% | 96% - 100% |
| *Site#1 tested two additional lots | | | | | |

# Overall Reproducibility Study

# II. Linearity/Assay Reportable Range: Not Applicable

# III. Level of Detection (LoD)

A Limit of Detection (LoD) study was performed to determine the analytical sensitivity of the IsoAmp HSV Assay using two representative strains of HSV-1 and two representative strains of HSV-2. Quantified (TCID50/mL) cultures of the HSV-1 and HSV-2 strains were serially diluted to five (5) concentrations in HSVnegative matrix pools and tested in replicates of ten (10) on three (3)

Negative pooled serum control

² Remel M4 transport media

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reagent lots. The observed LoD of a HSV strain was determined as the lowest concentration level that had a positivity rate of >95%. Since two (2) representative strains of HSV-1 and HSV-2 were used in the study, the higher LoD value was defined as the observed LoD for HSV-1 and HSV-2 respectively. Since the IsoAmp® HSV Assay does not differentiate viral types, the final assay LoD is defined as the higher of the HSV-1 and HSV-2 concentrations where 95% positivity was observed.

In addition, LoD confirmation studies were conducted to confirm the observed LoD for HSV-1 and HSV-2. The first confirmatory study included testing the four (4) representative HSV-1 and HSV-2 strains 20 times each at the corresponding observed LoD. Each strain was tested by three (3) reagent lots, and all four strains showed a positivity rate of 100%. In addition, twenty (20) HSV-1 and 20 HSV-2 clinical isolates were cultured and quantified in TCIDso/mL. Each isolate was diluted to the corresponding LoD in HSV-negative matrix and tested in triplicate. IsoAmp® HSV Assay was able to detect all 20 HSV-1 and 20 HSV-2 clinical isolates.

- a. HSV-1
The LoD for HSV-1 Strain 1 was determined to be 3.7 x 10° TCIDso/mL. At this concentration, 97% of samples were detected with a 95% Confidence Interval of 83.33% - 99.41%. The LoD for HSV-1 Strain 2 was determined to be 1.1 x 105 TCID50/mL. At this concentration, 100% of samples were detected with a 95% Confidence Interval of 88.65% - 100%. Therefore, the LoD for HSV-1 is 1.1 x 10° TCID50/mL.

| Strain 1<br>(TCID50/mL) | Positive/Total | Positivity Rate | 95% CI | |
|-------------------------|----------------|-----------------|--------|---------|
| 3.3 x 105 | 30/30 | 100% | 88.65% | 100.00% |
| 1.1 x 105 | 30/30 | 100% | 88.65% | 100.00% |
| 3.7 x 104 | 29/30 | 97% | 83.33% | 99.41% |
| 1.2 x 104 | 18/30 | 60% | 42.32% | 75.41% |
| 4.1 x 103 | 10/30 | 33% | 19.23% | 51.22% |
| Strain 2<br>(TCID50/mL) | Positive/Total | Positivity Rate | 95% CI | |
| 3.3 x 105 | 30/30 | 100% | 88.65% | 100.00% |
| 1.1 x 105 | 30/30 | 100% | 88.65% | 100.00% |
| 3.7 x 104 | 28/30 | 93% | 78.68% | 98.15% |
| 1.2 x 104 | 19/30 | 63% | 45.51% | 78.13% |
| 4.1 x 103 | 9/30 | 30% | 16.66% | 47.88% |

### b. HSV-2

The LoD for HSV-2 Strain 1 was determined to be 1.1 x 10° TCID50/mL. At this concentration, 100% of samples were detected with a 95% Confidence Interval of 88.65% - 100%. The LoD for HSV-2 Strain 2 was

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determined to be 3.7 x 103 TCID50/mL. At this concentration, 100% of samples were detected with a 95% Confidence Interval of 88.30% – 100%. Therefore, the LoD for HSV-2 is 1.1 x 104 TCIDso/mL.

| Strain 1<br>(TCID50/mL) | Positive/Total | Positivity Rate | 95% CI | |
|-------------------------|----------------|-----------------|--------|---------|
| 3.3 x 104 | 30/30 | 100% | 88.65% | 100.00% |
| 1.1 x 104 | 30/30 | 100% | 88.65% | 100.00% |
| 3.7 x 103 | 26/30 | 87% | 70.32% | 94.69% |
| 1.2 x 103 | 14/30 | 47% | 30.23% | 63.86% |
| 4.1 x 103 | 8/30 | 27% | 14.18% | 44.45% |
| Strain 2<br>(TCID50/mL) | Positive/Total | Positivity Rate | 95% CI | |
| 3.3 x 104 | 30/30 | 100% | 88.65% | 100.00% |
| 1.1 x 104 | 30/30 | 100% | 88.65% | 100.00% |
| 3.7 x 103 | 29/29 | 100% | 88.30% | 100.00% |
| 1.2 x 103 | 25/30 | 83% | 66.44% | 92.66% |
| 4.1 x 102 | 8/30 | 27% | 14.18% | 44.45% |

#### Assay LoD C.

Since the IsoAmp® HSV Assay does not differentiate viral types, the final assay LoD is defined as the higher of the HSV-1 and HSV-2 concentrations where 95% positivity was observed. The final assay LoD claim is 1.1 x 10-TCID50/mL.

#### IV. Cross Reactivity Testing (Analytical Specificity)

A cross-reactivity study was performed to determine if any organisms which may present with the same clinical symptoms as HSV, which are associated with bacterial vaginosis or which are commonly found in the genital track and oral area could give positive results with the IsoAmp® HSV Assay. Forty-eight (48) specificity panel members including purified DNA and cultured organisms were tested with the IsoAmp® HSV assay in triplicate following instructions in the Package Insert. No cross-reactivity was observed with any panel member tested at clinically significant concentrations.

| Organisms | Member Type (GD1,<br>QC2, IHC3) | Test Concentration |
|---------------------------------------------------------|---------------------------------|------------------------|
| Acinetobacter calcoaceticus var. anitratus (ATCC 51432) | IHC | $1.0 x 10^6$ CFU/mL |
| Acinetobacter lwoffi (ATCC 17925) | IHC | $1.0 x 10^7$ CFU/mL |
| Adenovirus 2 | QC | $1.0 x 10^6$ TCID50/mL |
| Bacteroides fragilis | QC | $1.0 x 10^7$ CFU/mL |
| Candida albicans (ATCC 14053) | IHC | $1.0 x 10^7$ CFU/mL |
| Organisms | Member Type (GD1,<br>QC2, IHC3) | Test Concentration |
| Candida glabrata | QC | 1.0 x 107 CFU/mL |
| Candida guilliermondii | QC | 1.0 x 107 CFU/mL |
| Candida krusei | QC | 1.0 x 106 CFU/mL |
| Candida lusitaniae | QC | 1.0 x 107 CFU/mL |
| Candida parapsilosis | QC | 1.0 x 107 CFU/mL |
| Candida tropicalis | QC | 1.0 x 107 CFU/mL |
| Chlamydia trachomatis LGV-I1434 | GD | 1.0 x 107 cp/mL |
| Cytomegalovirus | QC | 1.0 x 106 TCID50/mL |
| Enterobacter cloacae (ATCC 13047) | IHC | 1.0 x 107 CFU/mL |
| Enterovirus (Type 71) | QC | 1.0 x 105 TCID50/mL |
| Epstein-Barr Virus | GD | 1.0 x 106 cp/mL |
| Escherichia coli (ATCC 25922) | IHC | 1.0 x 107 CFU/mL |
| Fusobacterium nucleatum (ATCC 25586) | IHC | 1.0 x 107 CFU/mL |
| Gardnerella vaginalis (ATCC 14018) | IHC | 1.0 x 107 CFU/mL |
| Haemophilus ducreyi | QC | 8.5 x 105 CFU/mL |
| Human Herpes 6 virus (Z29 strain) | QC | 1.0 x 106 TCID50/mL |
| Human Herpes 7 virus (SB strain) | QC | 1.0 x 106TCID50/mL |
| Human papilloma virus 16 (HPV16) | GD | 1.0 x 108 cp/mL |
| Human papilloma virus 18 (HPV18) | GD | 1.0 x 105 cp/mL |
| Klebsiella pneumoniae | QC | 1.0 x 107 CFU/mL |
| Lactobacillus acidophilus Z048 | QC | 1.0 x 107 CFU/mL |
| Mobiluncus curtisii V125 [DSM 2711] | QC | 1.0 x 107 CFU/mL |
| Mobiluncus mulieris BV 64-5 | QC | 1.0 x 106 CFU/mL |
| Moraxella catarrhalis | QC | 1.0 x 107 CFU/mL |
| Mycoplasma hominis (ATCC 23114) | IHC | 1.0 x 107 CFU/mL |
| Neisseria gonorrhoeae (ATCC 21823) | IHC | 1.0 x 107 CFU/mL |
| Neisseria meningitides | QC | 1.0 x 107 CFU/mL |
| Prevotella melaninogenica | QC | 1.0 x 107 CFU/mL |
| Rubella virus | QC | 4.17 x 105 TCID50/mL |
| Simian Virus type 40 (SV40) | QC | 1.0 x 106 TCID50/mL |
| Staphylococcus aureus MRSA (ATCC 33591) | IHC | 1.0 x 107 CFU/mL |
| Staphylococcus aureus MSSA (ATCC 25923) | IHC | 1.0 x 107 CFU/mL |
| Staphylococcus epidermidis MRSE (ATCC700566) | IHC | 1.0 x 107 CFU/mL |
| Staphylococcus saprophyticus MRSE (ATCC 15305) | IHC | 1.0 x 107 CFU/mL |
| Streptococcus mitis clinical isolate | QC | 1.0 x 107 CFU/mL |
| Streptococcus mutans Z072 | QC | 1.0 x 106CFU/mL |
| Organisms | Member Type (GD1,<br>QC2, IHC3) | Test Concentration |
| Streptococcus pneumoniae | QC | 1.0 x 107 CFU/mL |
| Streptococcus pyogenes: (ATCC19615) | IHC | 1.0 x 107 CFU/mL |
| Streptococcus salivarius (ATCC BAA-1024) | IHC | 1.0 x 107 CFU/mL |
| Toxoplasma gondii | QC | 6.6 x 106 CFU/mL |
| Treponema pallidum | QC | 1.0 x 107 TP/mL |
| Trichomonas vaginalis | QC | 1.0 x 106 CFU/mL |
| Varicella-Zoster Virus (VZV) | GD | 1.0 x 106 cp/mL |

Genomic DNA

² Quantified Cultures

³ In-House Culture

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·

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# V. Interfering Substances

Potentially interfering substances i.e. viral transport media, substances that might be present in clinical samples, and organisms/cross reactive panel members listed under cross reactivity were tested to confirm that they did not interfere with the performance of the IsoAmp® HSV Assay.

All interference testing was carried out in the presence of HSV-1 and HSV-2 at three times the observed LoD (3 x LoD). All test runs were conducted in triplicate. Controls were tested with each run.

# a. Interfering Substances

Performance of the IsoAmp® HSV Assay was characterized in the presence of twenty-four (24) potentially interfering substances which could reasonably be expected to be present in genital and oral swab specimens. Interfering substances were tested at the highest ("worst case") concentration expected in clinical samples. The panel was also tested in triplicate in the absence of HSV to see if the potentially interfering substances interfere with the detection of the internal control. No interference was observed in the presence of the potential interfering substances tested.

| Substances (active ingredients) | Calculated Concentration |
|---------------------------------------------------|--------------------------|
| Whole blood with EDTA | 7% (v/v) |
| Female Urine | 7% (v/v) |
| Male Urine | 7% (v/v) |
| Acyclovir (Acycloguanosine) 10% | 7 mg/mL |
| Albumin | 3.3 mg/mL |
| Casein | 7 mg/mL |
| K-Y Brand Jelly | 7% (w/v) |
| Douche (Decyl Glucoside; Octoxynol-9) | 7% (v/v) |
| Contraceptive Jelly | 7% (w/v) |
| YeastGard (Phosphoricum Acidum 4X) | 7% (w/v) |
| Monistat 1 (Miconazole Nitrate cream (2%)) | 7% (w/v) |
| Vagisil Crème (Benzocaine (20%), Resorcinol (3%)) | 7% (w/v) |
| Monistat 3 (Miconazole Nitrate Cream (4%)) | 7% (w/v) |

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| Triconazole 1 (Tioconazole (300 mg) (6.5%)) | 7% (w/v) |
|---------------------------------------------------------------------------|----------|
| Balneol Hygienic Cleansing Lotion | 7% (w/v) |
| Clotrimazole 3 Vaginal Cream (Clotrimazole 100 mg (2%)) | 7% (w/v) |
| CVS Anti-Itch Cream (Benzocaine 5%; Benzalkonium Chloride 0.13%) | 7% (w/v) |
| Listerine Antiseptic Mouth Wash | 7% (v/v) |
| Abreva (Docosanol 10%) | 7% (w/v) |
| Carmex Cold Sore Lip Balm (Menthol (0.7%), Camphor (1.7%), Phenol (0.4%)) | 7% (w/v) |
| Releev cold sore treatment (Benzalkonium Chloride (0.13%)) | 7% (w/v) |
| Lip clear Lysine+ (Zinc Oxide (1.2%)) | 7% (w/v) |
| Toothpaste | 7% (w/v) |
| Buffy coat | 7% (v/v) |

# b. Viral Transport Media

The performance of the IsoAmp® HSV Assay was assessed with Remel M4, Remel M5, Remel M4RT, Bartels VTM, and BD Universal Viral Transport (UVT). Each medium was tested after spiking with HSV-1 and HSV-2 strain to a final concentration of approximately 3 x LoD to determine if the viral transport media interferes with the detection of HSV targets in positive samples. The media were tested in the absence of HSV-1 and HSV-2 (medium only) to see if the viral transport media interferes with the detection of the internal control in negative samples. There was no interference observed with the Remel M4, Remel M4RT, Remel M5, Bartels VTM, and BD UVT media for the detection of HSV-1 and HSV- 2 target or the internal control. M4, M4RT, M5, Bartels VTM, and BD UVT did not interfere with the detection of HSV-1 and HSV-target or the internal control.

Cross-Reactivity Panel Members C.

> The performance of the IsoAmp® HSV Assay was characterized by testing the organisms that were evaluated for analytical specificity and cross reactivity in the presence of HSV-1 and HSV-2 strains at 3xLoD separately to see if the presence of these organisms interferes with the detection of HSV target. Each panel member was tested in triplicate. None of the cross reactivity panel members interfered with the detection of HSV-1 and HSV-2 target.

# VI. Carry-Over and Cross Contamination

Carry-over/Cross Contamination Study was done only with HSV-1 target since both HSV-1 and HSV-2 share a single set of primers and probes for target amplification and detection. The HSV-1 Strain 1 was used directly without dilution. Viral transport media was used as the negative sample. Ten (10) replicates of negative sample together with assay controls were run by two (2) operators to confirm that negative samples generate a negative result 100% of the time. Five (5) replicates of high-concentration positive and negative samples were tested in a series, alternating sample types. All results were

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as expected. Negative samples tested were negative (10/10) and positive samples were positive (10/10).

- VII. Assay Cut-Off: Not Applicable
# Clinical Performance 7.0

The performance of the IsoAmp® HSV Assay was evaluated at five geographically diverse locations within the United States from 2010 - 2011. A total of nine hundred and ninetyfour (994) swab samples obtained from male and female genital and oral lesions were collected in Viral Transport Media (Remel M4, Remel M4RT, BD Universal Viral Transport and Bartels VTM) from the patient population ranging from <1 year to 92 years, and evaluated. Of the 994 specimens, 962 prospective samples and 32 retrospective samples were tested. Of the 962 prospective samples, 803 genital samples and 159 oral samples were tested. Of the 32 retrospective samples, 15 genital and 17 oral samples were tested at a single study site. Genital swab specimens were collected from vaginal, labial, penile and rectal lesions. Oral swab specimens were collected from lips, gums, and mouth.

The performance of the IsoAmp HSV Assay was compared with a gold standard/reference method i.e., Cell Culture based ELVIS® HSV ID/Typing Test System using an enzyme linked virus inducible system.

| | Reference Method | | | |
|-------------------|------------------|-----|-------------------------|-----|
| | POS | NEG | Total | |
| IsoAmp° HSV Assay | POS | 264 | 351 | 299 |
| | NEG | 82 | 496 | 504 |
| | Total | 272 | 531 | 803 |
| | | | | |
| | Value | | 95% Confidence Interval | |
| Sensitivity | 97.1% (264/272) | | 94.3 – 98.5% | |
| Specificity | 93.4% (496/531) | | 91.0 – 95.2% | |

# 1. Prospective Sample Data

- Genital Samples Only a.
^{* 35} samples were tested using bidirectional sequence analysis detected HSV target in 29 (6 HSV-1, 23 HSV-2) of the 35 discordant samples identified as HSV Positive by the Issay. Sequence analysis did not detect HSV in six (6) of the discordant samples.

² Eight (8) samples were tested using bidirectional sequence analysis did not detect HSV target in four (4) of the 8 samples identified as HSV Negative by the IsoAmp® HSV Assay. Sequence analysis did detect HSV in four (4) samples (2 HSV-2)

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#### b. Oral Samples Only

| | | Reference Method | | |
|------------------|----------------|------------------|-------------------------|-------|
| | | POS | NEG | Total |
| IsoAmp HSV Assay | POS | 45 | 14¹ | 59 |
| | NEG | 3² | 97 | 100 |
| | Total | 48 | 111 | 159 |
| | | | | |
| | Value | | 95% Confidence Interval | |
| Sensitivity | 93.8% (45/48) | | 83.2 - 97.9% | |
| Specificity | 87.4% (97/111) | | 79.9 - 92.3% | |

#### Retrospective Sample Data II.

All of the 32 retrospective samples, 15 genital and 17 oral samples were shown positive by both the IsoAmp HSV Assay and the reference assay.

#### 8.0 Statement of Supporting Data

The results of the analytical and clinical performance studies submitted in this premarket notification are complete and demonstrate that the IsoAmp® HSV Assay is substantially equivalent to the predicate device.

^{ี 14} samples were tested using bidirectional sequence analysis detected HSV target in 13 [12 HSV-2] of the 14 discordant samples identified as HSV Positive by the IsoAmp® HSV Assay. Sequence analysis did not detect HSV in one (1) of the discordant samples.

⁷ Three (3) samples were tested using bidirectional sequence analysis did not detect HSV target in two (2) of the 3 samples identified as HSV Negative by the IsoAmp® HSV Assay. Sequence analysis did detect HSV in one (1) samples (1 HSV-1)

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Image /page/11/Picture/1 description: The image shows the logo for the U.S. Department of Health and Human Services. The logo consists of a circular border with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the circumference. Inside the circle is an abstract symbol resembling a stylized human figure with outstretched arms, possibly representing care or protection.

Food and Drug Administration 10903 New Hampshire Avenue Silver Spring, MD 20993

BioHelix Corporation c/o Fran White Regulatory Consultant 500 Cummings Center Suite 5550 Beverly, MA 01915

27 230

Re: K111951

Trade/Device Name: IsoAmp® HSV Assay Regulation Number: 21 CFR \$866.3305 Herpes Simplex Virus Nucleic Acid Amplification Assay Regulation Name: Regulatory Class: Class II Product Code: 000 Dated: July 6, 2011 Received: July 8, 2011

Dear Ms. White:

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.

If your device is classified (see above) into class II (Special Controls), it may be subject to such additional controls. Existing major regulations affecting your device can be found in Title 21, Code of Federal Regulations (CFR), Parts 800 to 895. 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 medical device-related adverse events) (21 CFR 803); and good manufacturing practice

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requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820). This letter will allow you to begin marketing your device as described in your Section 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.

If you desire specific advice for your device on our labeling regulation (21 CFR Parts 801 and 809), please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at [phone]. 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 Small Manufacturers, International and Consumer Assistance at its toll-free number [phone] or [phone] or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html.

Sincerely yours,

Wine Salt for

Sally A. Hojvat, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

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# Indications for Use

510(k) Number (if known): K11951

lsoAmp® HSV Assay Device Name:

Indications for Use:

The IsoAmp® HSV Assay is an in vitro diagnostic test for the direct, qualitative detection of herpes simplex virus (HSV-1 & HSV-2) DNA in male and female genital and oral lesions. The test is intended for use as an aid in diagnosis of HSV infection in symptomatic patients.

Warning: The IsoAmp® HSV Assay is not FDA cleared for use with cerebrospinal fluid (CSF). The assay does not provide specific typing information to differentiate HSV-1 and HSV-2. The assay is not intended to be used for prenatal screening.

Prescription Use _____________________________________________________________________________________________________________________________________________________________ × (Part 21 CFR 801 Subpart D)

AND/OR

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

(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE OF NEEDED)

Concurrence of CDRH, Office of In Vitro Diagnostic Device Evaluation and Safety (OIVD)

Tiawad, Feldsly

Division Sign-Off Office of In Vitro Diagnostic Device Evaluation and Safety

510(k) 111951

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