ELVIS HSV ID AND D3 TYPING TEST SYSTEM

K091753 · Diagnostic Hybrids, Inc. · GQL · Aug 28, 2009 · Microbiology

Device Facts

• Record ID: K091753
• Device Name: ELVIS HSV ID AND D3 TYPING TEST SYSTEM
• Applicant: Diagnostic Hybrids, Inc.
• Product Code: GQL · Microbiology
• Decision Date: Aug 28, 2009
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3305
• Device Class: Class 2

Intended Use

The ELVIS®HSV ID and D3 Typing Test System provides Cells, Replacement Medium and Test Reagents for the culture, qualitative identification and typing of herpes simplex virus (HSV) from cutaneous or mucocutaneous specimens as an aid in the diagnosis of HSV type 1 (HSV-1) and HSV type 2 (HSV-2) infections. The performance characteristics of this assay have not been established for antiviral therapy, prenatal monitoring or use with cerebral spinal fluid specimens.

Device Story

System uses genetically engineered Baby Hamster Kidney (BHK) cells to culture HSV from patient cutaneous/mucocutaneous specimens. Infected cells express endogenous beta-galactosidase; HSV-1/HSV-2 infection also produces type-specific proteins. Procedure involves specimen inoculation into cell monolayers, incubation, and staining. HSV-1 detected via non-labeled monoclonal antibodies; HSV-2 detected via fluorescein-labeled monoclonal antibodies. Results visualized via fluorescence microscopy; blue-stained cells indicate HSV presence. Used in clinical laboratories by trained personnel. Output aids physician diagnosis of HSV-1/HSV-2 infections. Benefits include rapid, specific viral identification to guide clinical management.

Clinical Evidence

Method comparison study across three sites with 719 clinical specimens. Compared subject device to predicate. Results: PPA for HSV isolation 99.6% (250/251); NPA 98.9% (463/468). HSV-2 typing PPA 99.3% (145/146); NPA 94.2% (98/104). HSV-1 typing PPA 100% (32/32); NPA 87.5% (7/8). Analytical studies included reproducibility (100% concordance), limit of detection (comparable to predicate), and specificity (no cross-reactivity with common respiratory viruses or bacteria, except S. aureus protein A binding).

Technological Characteristics

System components: genetically engineered BHK cells, replacement medium, test reagents (Solution 1 fixative, Solution 2T chromogenic/MAb stain, Solution 3 secondary stain). Detection: β-galactosidase enzyme activity (chromogenic) and immunofluorescence (FITC-labeled MAbs). Format: shell vials or multi-well plates. Centrifugation-enhanced inoculation. Manual microscopic examination. No electronic connectivity or software algorithms.

Indications for Use

Indicated for qualitative identification and typing of HSV-1 and HSV-2 from cutaneous or mucocutaneous specimens in patients with clinical suspicion of HSV infection. Not for use in antiviral therapy monitoring, prenatal monitoring, or CSF specimens.

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

• ELVIS®HSV ID/Typing Test System (k971662)

Submission Summary (Full Text)

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K091753

# ELVIS®HSV ID & D3 Typing Test System

08/14/2009 Page 1 of 14

# 510(k) Summary

Applicant:

AUG 2 8 2009

DIAGNOSTIC HYBRIDS, INC. 1055 East State Street Suite 100 Athens, OHIO 45701

#### Contact Information:

Ronald H. Lollar, Senior Director Product Realization, Management, and Marketing 1055 East State Street Suite 100 Athens, Ohio 45701 740-589-3300 - Corporate number 740-589-3373 - Desk phone [phone] - Fax [e-mail]

### Date of preparation of 510(k) summary:

June 12, 2009

#### Device Name:

Trade name – ELVIS®HSV ID and D3 Typing Test System Common name - HSV Culture and Typing Classification name -- Antisera, fluorescent, herpesvirus hominis 1,2 Product Code - GQL Regulation - 21 CFR Sec. 866.3305 Herpes simplex virus serological assays; Panel - Microbiology (83)

### Legally marketed devices to which equivalence is claimed:

ELVIS®HSV ID/Typing Test System (k971662)

Intended Use:

The ELVIS®HSV ID and D3 Typing Test System provides Cells, Replacement Medium and Test Reagents for the culture, qualitative identification and typing of Herpes simplex virus (HSV) from cutaneous or mucocutaneous specimens collected from patients with clinical suspicion of HSV infection. The performance characteristics of this assay have not been established for antiviral therapy, prenatal monitoring or CSF specimens.

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### Device Description:

The ELVIS®HSV ID and D3 Typing Test System provides Cells, Replacement Medium and Test Reagents for the culture, qualitative identification and typing of herpes simplex virus (HSV) from cutaneous or mucocutaneous specimens as an aid in the diagnosis of HSV type 1 (HSV-1) and HSV type 2 (HSV-2) infections. The performance characteristics of this assay have not been established for antiviral therapy, prenatal monitoring or use with cerebral spinal fluid specimens.

ELVIS HSV Cells are genetically engineered Baby Hamster Kidney (BHK) cells, which, when infected with either HSV-1 or HSV-2, are induced to generate and accumulate an endogenous, intracellular bacterial enzyme, ßgalactosidase. Other related viruses (e.g., Varicella zoster) are not capable of inducing the formation of this enzyme. HSV infection of the ELVIS®HSV Cells also results in the formation of HSV-type-specific proteins. The presence of these proteins can be detected microscopically when fluorescent labeled HSV-type-specific antibodies are used. The two Type 1 monoclonal antibodies used in ELVIS® are directed against specific to epitopes on the HSV-1 protein. The three Type 2 monoclonal antibodies are directed against the HSV-2 glycoproteins C, G and a recombinant glycoprotein G that occur in the cytoplasm of infected cells.

The ELVIS®HSV ID and D2 Typing Test System consists of:

- 1. ELVIS®HSV Cells: The ELVIS®HSV Cells have a routine use period of 7 days from customer receipt while all other components have a shelflife of months (see expiration date on label of each component). ELVIS® HSV Cells are provided as 75% to 95% confluent monolavers in shell-vials with or without coverslips, or in multi-well plates with or without coverslips, and up to 24 monolayers per plate. Each monolayer is covered by at least 0.75-mL of Eagle's Minimum Essential Medium (EMEM) with fetal bovine serum (FBS), penicillin, and streptomycin. Cells are characterized by isoenzyme analysis and have been tested and found free of Mycoplasma spp. and other adventitious organisms.
- 2. ELVIS HSV Replacement Medium: Sterile EMEM containing FBS, Penicillin, Streptomycin and Amphotericin B. ELVIS®HSV Replacement Medium is for use with ELVIS®HSV Shell-Vials and Multi-well Plates.
- 3. ELVIS® HSV Solution 1 (Cell Fixative): an aqueous acetone solution.
- 4. ELVIS® HSV Solution 2T (Staining Buffer): A diluted solution of X-Gal (5-Bromo-4-Chloro-3-Indolyl-B-D-Galactopyranoside), N.N-Dimethylformamide, iron, sodium and magnesium salts, fluorescein-labeled HSV-2-specific murine MAbs (directed against

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HSV-2 glycoproteins C, G, and a recombinant glycoprotein G) and nonlabeled HSV-1-specific murine MAbs (specific to epitopes on the HSV-1 protein UL42), penicillin, streptomycin, bovine serum albumin and Evans Blue in an aqueous, buffered solution.

- 5. ELVIS®HSV Solution 3: An aqueous, stabilized, buffered solution containing fluorescein-labeled, affinity purified goat-anti-mouse IgG antibody and Evans Blue with sodium azide as preservative.
- ELVIS®HSV Mounting Fluid (Buffered Glycerol): Aqueous, stabilized, 6. buffered glycerol (pH 7.3 +/- 0.5), containing sodium azide as preservative.
- 7. 40X PBS Concentrate. 25-mL: One bottle of a 40X PBS concentrate consisting of 0.4% sodium azide (0.1% sodium azide after dilution to 1X using de-mineralized water).

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ELVIS®HSV ID & D3 Typing Test System

8/25/2009 Section 05, Page 4 of 14

Image /page/3/Figure/3 description: This image is a flowchart outlining the ELVIS® Procedure. The flowchart is divided into four main sections: SET UP, VIRUS AMPLIFICATION, COLOR DEVELOPMENT, and EXAMINATION FOR RESULT. The SET UP section involves pre-incubating ELVIS HSV cell cultures for 2 to 16 hours at 35°C to 37°C, processing the specimen, removing culture maintenance medium, adding replacement medium, and inoculating cells with prepared specimen material. The EXAMINATION FOR RESULT section involves determining if blue-stained cells are present, and if not, the specimen is negative for HSV, and if so, the specimen is positive for HSV and the procedure continues.

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### Intended Use:

The ELVIS® HSV ID and D3 Typing Test System provides Cells, Replacement Medium and Test Reagents for the culture, qualitative identification and typing of herpes simplex virus (HSV) from cutaneous or mucocutaneous specimens as an aid in the diagnosis of HSV type 1 (HSV-1) and HSV type 2 (HSV-2) infections. The performance characteristics of this assay have not been established for antiviral therapy, prenatal monitoring or use with cerebral spinal fluid specimens.

| Table 5.1: Subject Device and Predicate Device Characteristics | | |
|----------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------|
| Similarities | | |
| Item | Subject Device | Predicate Device |
| Intended Use | The <i>ELVIS®HSV</i> ID and D3<br>Typing Test System provides<br>Cells, Replacement Medium and<br>Test Reagents for the culture,<br>qualitative identification and<br>typing of <i>Herpes simplex</i> virus<br>(HSV). | Same |
| Assay Format | Shell-vials or Multi-well plates | Same |
| Assay principle | Genetically engineered Baby<br>Hamster Kidney (BHK) cells,<br>which, when infected with either<br>HSV-1 or HSV-2, are induced to<br>generate and accumulate an<br>endogenous, intracellular<br>bacterial enzyme, β-<br>galactosidase. | Same |
| Labeling Method | Direct Method –<br>Using fluorescein isothiocyanate<br>(FITC) to label HSV-2 Specific<br>monoclonal antibodies, and goat-<br>anti-mouse IgG antibody | Same |
| Differences | | |
| Item | Subject Device | Predicate Device |
| Monoclonal<br>Antibodies<br>(MAbs) | HSV-1: non-labeled specific to<br>epitopes on the HSV-1 protein<br>UL42 | HSV-1: non-labeled specific to<br>HSV-1 viral protein occurring in<br>the nuclei of infected cells and an<br>HSV-1 glycoprotein C |
| | HSV-2: FITC labeled specific | HSV-2: FITC labeled specific for |

### Technological Characteristics, Compared to Predicate Device:

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| Table 5.1: | Subject Device and Predicate Device Characteristics | |
|------------|----------------------------------------------------------------|------------------------------|
| | for HSV-2 glycoproteins C, G, and a recombinant glycoprotein G | HSV-2 glycoproteins C, and G |

## Performance Testing - Non-Clinical

# A. Analytical Sensitivity

Analytical detection limits for HSV-1 and HSV-2 were addressed with results reported in numbers of blue staining cells per cell monolayer. Each master stock (~1e7-TCID50 per mL) virus preparation underwent a series of ten-fold dilutions, which were subsequently inoculated into a 96-well ELVIS®HSV cell culture plate. The plates were centrifuged at 700xg for 60 minutes, and then incubated at 35°C to 37°C for 17-hours. Each well was stained with the subject and predicate devices then examined at 200X magnification and the number of blue staining cells counted. Table 5.2. below lists the results for each virus strain tested.

| Table 5.2: Limit of Detection compared between ELVIS Subject (D³ ELVIS) and | | | |
|-----------------------------------------------------------------------------|--------------------|----------------------------|-------------------------|
| Predicate (Current ELVIS Kit Formulation) Typing Systems | | | |
| Virus strain | Virus per Inoculum | ELVIS Predicate | ELVIS Subject |
| HSV-1 Strain F<br>ATCC VR-733 | 65-TCID50 | 74, 67, 65, 69, 70, 64 | 76, 70, 63, 68, 72, 71 |
| | 6.5-TCID50 | 9, 8, 11, 7, 7, 12 | 10, 9, 9, 11, 7, 13 |
| | 0.65-TCID50 | 1, 2, 1, 1, 3, 3 | 3, 2, 4, 3, 1, 1 |
| | 0.065-TCID50 | 0, 0, 3, 1, 1, 0 | 0, 0, 1, 2, 0, 0 |
| | 0.0065-TCID50 | 0, 0, 0, 0, 0, 0 | 0, 0, 0, 0, 0, 0 |
| HSV-1 CW0H0062<br>Clinical Isolate<br>Passage 2 | 85-TCID50 | 70, 79, 75, 72, 80, 67 | 82, 77, 72, 65, 76, 85 |
| | 8.5-TCID50 | 10, 7, 7, 6, 9, 6 | 11, 10, 8, 6, 7, 7 |
| | 0.85-TCID50 | 0, 1, 3, 0, 0, 1, 0 | 2, 0, 0, 0, 2, 2 |
| | 0.085-TCID50 | 0, 0, 0, 0, 1, 0 | 1, 0, 0, 0, 1, 0 |
| | 0.0085-TCID50 | 0, 0, 0, 0, 0, 0 | 0, 0, 0, 0, 0, 0 |
| HSV-1 CWOH0085<br>Clinical Isolate<br>Passage 2 | 60-TCID50 | 39, 47, 52, 41, 42, 48 | 46, 48, 37, 42, 47, 50 |
| | 6.0-TCID50 | 6, 10, 11, 8, 7, 15 | 7, 14, 9, 8, 11, 7 |
| | 0.6-TCID50 | 2, 0, 2, 0, 0, 1 | 1, 1, 0, 0, 0, 1 |
| | 0.06-TCID50 | 0, 0, 0, 0, 0, 0 | 0, 0, 0, 0, 0, 0 |
| | 100-TCID50 | 92, 102, 95, 91, 97,<br>90 | 95, 96, 97, 98, 89, 103 |
| HSV-2 G Strain<br>ATCC VR-734 | 10-TCID50 | 12, 11, 17, 9, 9, 10 | 12, 12, 7, 16, 13, 12 |
| | 1.0-TCID50 | 3, 2, 1, 1, 3, 4 | 5, 1, 2, 2, 1, 3 |
| | 0.1-TCID50 | 0, 1, 0, 1, 0, 0 | 1, 0, 0, 0, 1, 1 |
| | 0.01-TCID50 | 0, 0, 0, 0, 0, 0 | 0, 0, 0, 0, 0, 0 |

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ELVIS®HSV ID & D3 Typing Test System

8/25/2009 Section 05, Page 7 of 14

| Table 5.2: Limit of Detection compared between ELVIS Subject (D³ ELVIS) and | | | |
|-----------------------------------------------------------------------------|---------------|------------------------|------------------------|
| Predicate (Current ELVIS Kit Formulation) Typing Systems | | | |
| | 80-TCID50 | 70, 67, 73, 78, 70, 62 | 76, 77, 64, 80, 70, 69 |
| HSV-2 CWOH0082 | 8.0-TCID50 | 8, 7, 10, 11, 6, 5 | 7, 8, 14, 11, 11, 9 |
| Clinical Isolate | 0.8-TCID50 | 1, 0, 3, 3, 2, 2, 1 | 2, 1, 1, 3, 1, 0 |
| Passage 2 | 0.08-TCID50 | 0, 0, 1, 0, 0, 0 | 0, 1, 0, 0, 0, 0 |
| | 0.008-TCID50 | 0, 0, 0, 0, 0, 0 | 0, 0, 0, 0, 0, 0 |
| | 55-TCID50 | 53, 61, 55, 62, 67, 65 | 70, 62, 55, 57, 53, 59 |
| HSV-2 CWOH0091 | 5.5-TCID50 | 3, 7, 7, 9, 2, 4 | 4, 4, 7, 8, 10, 3 |
| Clinical Isolate | 0.55-TCID50 | 1, 0, 0, 2, 2, 1 | 3, 1, 0, 0, 2, 2 |
| Passage 2 | 0.055-TCID50 | 0, 0, 0, 1, 0, 0 | 1, 0, 0, 0, 0, 0 |
| | 0.0055-TCID50 | 0, 0, 0, 0, 0, 0 | 0, 0, 0, 0, 0, 0 |

In this study, the detection limit for the test is defined as the lowest inoculum level at which positive wells (i.e., containing blue staining cells) are observed, in terms of TCID50. The results presented in Table 5.2 above indicate that detection limit for both subject and predicate devices averages between 0.65and 8.5-TCID50 for HSV-1 and 0.1 and 8.0-TCID50 for HSV-2 depending on the strain.

### B. Cross Reactivity

The specificity of the MAbs used in the device was assessed using the organisms listed in Table 5.3. The subject device Solution 2T at 2X concentration was tested in duplicate on the prepared slides. After 1-hour at 37°C, the slides were rinsed with PBS and the subject device Solution 3 secondary stain was added and incubated at 37℃ for 15 minutes. After rinsing and applying Mounting Fluid, the slides were examined at 400X using a fluorescence microscope.

| Organism | Strain or Type | ELVIS HSV Typing<br>Reagent at 2X<br>concentration<br>[Positive (+) or<br>Negative (-) for<br>Reactivity] | Concentrations of<br>targets (viruses:<br>TCID50 inoculum<br>level; bacteria:<br>CFU) |
|-------------------|-------------------------|-----------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------|
| Viruses | | | |
| Adenovirus | Type 1 | - | 1000-TCID50 |
| | Type 3 | - | 1000-TCID50 |
| | Type 5 | - | 1000-TCID50 |
| | Type 6 | - | 1000-TCID50 |
| | Type 7 | - | 1000-TCID50 |
| | | | |
| | Type 8 | - | 1000-TCID50 |
| | Type 10 | - | 1000-TCID50 |
| | Type 13 | - | 1000-TCID50 |
| | Type 14 | - | 1000-TCID50 |
| | Type 18 | - | 1000-TCID50 |
| | Type 31 | - | 1000-TCID50 |
| | Aichi (H3N2) | - | 1000-TCID50 |
| | Mal (H1N1) | - | 1000-TCID50 |
| | Hong Kong (H3N2) | - | 1000-TCID50 |
| | Denver (H1N1) | - | 1000-TCID50 |
| Influenza A | Port Chalmers<br>(H3N2) | - | 1000-TCID50 |
| | Victoria (H3N2) | - | 1000-TCID50 |
| | New Jersey<br>(HSWN1) | - | 1000-TCID50 |
| | WS (H1N1) | - | 1000-TCID50 |
| | PR (H1N1) | - | 1000-TCID50 |
| | Hong Kong | - | 1000-TCID50 |
| | Maryland | - | 1000-TCID50 |
| | Mass | - | 1000-TCID50 |
| Influenza B | GL | - | 1000-TCID50 |
| | Taiwan | - | 1000-TCID50 |
| | JH-001 Isolate | - | 1000-TCID50 |
| | Russia | - | 1000-TCID50 |
| | Long | - | 1000-TCID50 |
| RSV | Wash | - | 1000-TCID50 |
| | 9320 | - | 1000-TCID50 |
| Parainfluenza 1 | C-35 | - | 1000-TCID50 |
| Parainfluenza 2 | Greer | - | 1000-TCID50 |
| Parainfluenza 3 | C-243 | - | 1000-TCID50 |
| Parainfluenza 4 | M-25 | - | 1000-TCID50 |
| Parainfluenza 4b | CH-19503 | - | 1000-TCID50 |
| CMV | AD169 | - | Control Slide |
| Varicella-zoster | Webster | - | Control Slide |
| Echovirus 7 | ODH-594684 | - | Control Slide |
| Coxsackievirus A9 | ODH-36685 | - | Control Slide |
| Coxsackievirus B2 | ODH-185 | - | Control Slide |
| Enterovirus 71 | ODH 02-89 | - | Control Slide |
| Bacteria* | | | |

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ELVIS®HSV ID & D³ Typing Test System

Section 05, Page 8 of 14

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ELVIS®HSV ID & D3 Typing Test System

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|--------------------------|
| Section 05. Page 9 of 14 |

| Acinetobacter<br>calcoaceticus | | | 3.6x109 CFU |
|----------------------------------|--------|----|---------------|
| Bordetella bronchiseptica | | | 1.1x1010 CFU |
| Bordetella pertussis | | | 4.3x109 CFU |
| Chlamydia trachomatis | LGV-II | | Control Slide |
| Corynebacterium<br>diphtheriae | | | 5.7x107 CFU |
| Escherichia coli | | | 7.5x108 CFU |
| Haemophilis influenzae<br>type A | | | 4.1x109 CFU |
| Klebsiella pneumoniae | | | 1.2x109 CFU |
| Moraxella cartarrhalis | | | 1.2x1010 CFU |
| Mycoplasma hominis | | | 3.5x1010 CFU |
| Mycoplasma orale | | | 6.6x109 CFU |
| Mycoplasma pneumoniae | | | 7.9x1010 CFU |
| Mycoplasma salivarium | | | 7.7x108 CFU |
| Proteus mirabilis | | | 3.6x109 CFU |
| Pseudomonas aeruginosa | | | 1.0x108 CFU |
| Salmonella enteriditis | | | 8.7x109 CFU |
| Salmonella typhimurium | | | 7.5x109 CFU |
| Staphylococcus aureus | | +† | 6.3x109 CFU |
| Streptococcus agalactiae | | | 5.5x108 CFU |
| Streptococcus pneumoniae | | | 6.7x109 CFU |
| Streptococcus pyogenes | | | 6.9x109 CFU |
| Yeast* | | | |
| Candida glabrata | | | 1.6x106 CFU |

* Turbidity or a color change to yellow indicates possible bacterial contamination and may render a test result unreliable, due either to a technical contamination during the culture setup or to a contaminated specimen. We recommend the original specimen be filtered and re-cultured.

* Light background fluorescent staining may occur with specimens contaminated with Staphylococcus aureus strains containing large amounts of protein A binds to the Fc portions of the conjugated antibodies. Such binding can be distinguished from viral antigen binding on the basis of morphology, e.g., S. aureus-bound fluorescence appears as small (~1 micron diameter), bright dots.

### C. Reproducibility Testing

The reproducibility of the device was assessed by creating ten panels of proficiency-level frozen virus suspensions. The panels were processed at each testing site. Each panel was inoculated and stained once according to the ELVIS®HSV ID and D3 Typing Test System instructions for use. Two panels per day were tested on separate plates for 5-days (10 total runs).

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Panel members were manufactured by diluting high-titered master stocks. The dilutions were made with the same lot of EMEM with 10% Fetal Bovine Serum used as the negative control. These dilutions were frozen at -70°C and sent to the testing labs. The dilution's titer was confirmed pre- and post freezing and found to fall within the expected infectivity range for the study: low level should exhibit less than 10% of the cells showing fluorescence; high level should exhibit greater than 10% but less than 50% of the cells showing fluorescence.

| Table 5.4: Panel Member Discriptions | |
|--------------------------------------|---------------------------------------------|
| Panel Member | Description |
| HSV-1 low level | SF029* lab adapted QC strain; 200 TCID50/mL |
| HSV-1 high level | SF029 lab adapted QC strain; 1000 TCID50/mL |
| HSV-2 low level | SF028† lab adapted QC strain; 200 TCID50/mL |
| HSV-2 high level | SF028 lab adapted QC strain; 1000 TCID50/mL |
| Negative | EMEM with 10% Fetal Bovine Serum |

*Isolate confirmed as HSV-1 by 2 FDA cleared IVD devices *Isolate confirmed as HSV-2 by 2 FDA cleared IVD devices

Table 5.5 presents the daily results from each panel member at each site.

| Table 5.5: Daily Results | | | | | | | | | | | |
|--------------------------|--------|-------|-------|-------|-------|------------|-------|-------|------------|-------|-------|
| Panel<br>Member | | Day 1 | Day 2 | Day 3 | Day 4 | Day 5 | | | | | |
| | | Run 1 | Run 2 | Run 1 | Run 2 | Run 1 | Run 2 | Run 1 | Run 2 | Run 1 | Run 2 |
| HSV-1<br>low level | Site 1 | +/- | +/- | +/- | +/- | 1+ | +/- | +/- | 1+ | 1+ | +/- |
| | Site 2 | +/- | 1+ | 1+ | 1+ | +/- | 1+ | 1+ | 1+ | +/- | 1+ |
| | Site 3 | 1+ | 1+ | 1+ | 1+ | 1+ | 1+ | 1+ | 1+ | +/- | 1+ |
| HSV-1<br>high level | Site 1 | 1+ | 1+ | 1+ | 1+ | 1 to<br>2+ | 1+ | 1+ | 1+ | 1+ | 1+ |
| | Site 2 | 1+ | 1+ | 1+ | 2+ | 1+ | 1+ | 3+ | 2+ | 1+ | 2+ |
| | Site 3 | 2+ | 2+ | 2+ | 2+ | 2+ | 2+ | 2+ | 3+ | 1+ | 2+ |
| HSV-2<br>low level | Site 1 | 1+ | +/- | 1+ | +/- | 1+ | +/- | +/- | +/- | 1+ | +/- |
| | Site 2 | +/- | 1+ | 1+ | 1+ | 1+ | 1+ | 2+ | 1 to<br>2+ | 1+ | 2+ |
| | Site 3 | 1+ | 1+ | 1+ | 1+ | 1+ | 1+ | 1+ | 1+ | +/- | 1+ |
| HSV-2<br>high level | Site 1 | 1+ | +/- | 1+ | +/- | 1+ | 1+ | 1+ | 1+ | 1+ | 1+ |
| | Site 2 | 2+ | 2+ | 2+ | 2+ | 2+ | 1+ | 3+ | 3+ | 3+ | 2+ |
| | Site 3 | 2+ | 3+ | 3+ | 3+ | 2+ | 3+ | 2+ | 3+ | 1+ | 2+ |
| Negative | Site 1 | NEG | NEG | NEG | NEG | NEG | NEG | NEG | NEG | NEG | NEG |
| | Site 2 | NEG | NEG | NEG | NEG | NEG | NEG | NEG | NEG | NEG | NEG |

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NEG | NEG Site 3 NEG NEG | NEG | NEG | NEG NEG NEG | NEG

The presence of HSV was reported in 100% (120/120) of the wells in which infected cells were present and the expected type was reported 100% (60/60) for HSV-1 and 100% (60/60) for HSV-2. The absence of HSV was reported in 100% (30/30) of the vials in which no virus was present. Controls performed as expected during each run.

| | Table 5.6: Reproducibility Study Summary Results | | | | | | |
|--------|--------------------------------------------------|-----------------------------|-----------------------------|-----------------------------|-----------------------------|---------------------------|----------------------|
| | Panel<br>Member | HSV-1<br>SF029<br>Low Level | HSV-1<br>SF029<br>Mid Level | HSV-2<br>SF028<br>Low Level | HSV-2<br>SF028<br>Mid Level | Negative<br>Control | Total %<br>Agreement |
| | Concentration | 200<br>TCID50/<br>mL | 1000<br>TCID50/<br>mL | 200<br>TCID50/<br>mL | 1000<br>TCID50/m<br>L | Non-<br>infected<br>cells | |
| Site 1 | Agreement with<br>Expected result | 10/10<br>(100%) | 10/10<br>(100%) | 10/10<br>(100%) | 10/10<br>(100%) | 10/10<br>(100%) | 50/50<br>(100%) |
| Site 2 | Agreement with<br>Expected result | 10/10<br>(100%) | 10/10<br>(100%) | 10/10<br>(100%) | 10/10<br>(100%) | 10/10<br>(100%) | 50/50<br>(100%) |
| Site 3 | Agreement with<br>Expected result | 10/10<br>(100%) | 10/10<br>(100%) | 10/10<br>(100%) | 10/10<br>(100%) | 10/10<br>(100%) | 50/50<br>(100%) |
| | Total Agreement<br>with Expected<br>result | 30/30<br>(100%) | 30/30<br>(100%) | 30/30<br>(100%) | 30/30<br>(100%) | 30/30<br>(100%) | 150/150<br>(100%) |
| | 95% CI | 88.4%-<br>100% | 88.4%-<br>100% | 88.4%-<br>100% | 88.4%-100% | 88.4%-<br>100% | 97.6%-100% |

## Performance Testing - Clinical

Studies were performed at three locations using 735 specimens submitted, April through May, 2009, for HSV culture. The number of specimens cultured at each of the three sites: Study site 1 - 299 specimens; Study site 2 - 136 specimens; and Study site 3 - 300 specimens. The specimens were cultured in duplicate and stained concurrently with both devices. The data generated by each site was similar and has been combined for presentation. Of these 735 specimens, 16 were excluded from the final analysis for the reasons listed in Table 5.7.

| Table 5.7: Combined Study Sites Rejected Specimens/Samples | |
|------------------------------------------------------------|----|
| Exclusion criteria - Toxic to cell culture | 13 |
| Exclusion criteria - Contaminated | 3 |
| Grand Total | 16 |

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| Table 5.8: Combined Study Sites - Age and Gender Distribution<br>(720 Specimens) | | | |
|----------------------------------------------------------------------------------|---------------------------------------------------------|---------|---------|
| Age Range | Values are # Positive (based on Subject Device) / Total | | |
| | Male | Female | Total |
| 0 to 1 month | 0/9 | 1/9 | 1/18 |
| >1 month to 2 years | 0/1 | 0/1 | 0/2 |
| >2 to 12 years | 1/7 | 4/7 | 5/14 |
| >12 to 21 years | 4/22 | 54/110 | 58/132 |
| 22 to 30 years | 9/34 | 71/146 | 80/180 |
| 31 to 40 years | 10/37 | 44/121 | 54/158 |
| 41 to 50 years | 8/22 | 18/64 | 26/86 |
| 51 to 60 years | 3/14 | 15/50 | 18/64 |
| >60 years | 3/18 | 9/47 | 12/65 |
| Unknown age | 0/0 | 0/0 | 0/0 |
| Grand Total | 38/165 | 216/555 | 254/719 |

Table 5.8 shows the age and gender distribution for individuals included in the Study:

Table 5.9 shows the specimen source distribution for the Study:

| Source | Total<br>Specimens | Unknown +/- | Genital +/- | Penis +/- | Vaginal +/- | Labia +/- | Cervical +/- | Wound +/- | Perineum * +/- | Vulva +/- | Urethra +/- | Lesion +/- | Face** +/- | Mouth ** +/- | Skin *+/- | Bartholin Cyst +/- |
|------------------------------------------------------|--------------------|-------------|-------------|-----------|-------------|-----------|--------------|-----------|----------------|-----------|-------------|------------|------------|--------------|-----------|--------------------|
| | 254/<br>719 | 66/<br>175 | 18/<br>50 | 14/<br>44 | 45/<br>105 | 23/<br>47 | 18/<br>50 | 0/4 | 16/<br>40 | 23/<br>66 | 0/<br>12 | 5/<br>14 | 4/<br>32 | 9/<br>37 | 13/<br>42 | 1/1 |
| * Perineum: anal, groin, buttock, perianal, tailbone | | | | | | | | | | | | | | | | |
| ** Mouth: mouth, lip, throat, NP Wash, Tongue | | | | | | | | | | | | | | | | |

Skin: skin, arm, back, breast, finger, foot, leg, thigh, breast, abdomen, hand

** Face: cheek, chin, eye, nasal

.

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Table 5.10 shows the comparison of the Subject device with the Predicate device for the isolation and detection of HSV at Study Sites Combined:

| Table 5.10: Combined Study Sites - Subject Device compared to<br>Predicate Device for the Isolation of HSV | | | |
|------------------------------------------------------------------------------------------------------------|------------|-----------------------------------------------------|-----------------|
| Specimen (719 specimens) | | Predicate Device<br>(Current ELVIS Kit Formulation) | |
| | | Pos | Neg |
| Subject Device (D3 ELVIS) | Pos | 250 | 5 |
| | Neg | 1 | 463 |
| Positive Percent Agreement (PPA) | | 99.6% (250/251) | |
| | 95% CI-PPA | 97.8 - 100% | |
| Negative Percent Agreement (NPA) | | | 98.9% (463/468) |
| | 95% CI-NPA | | 97.5 – 99.7% |

Table 5.11 shows the comparison of the Subject device with the Predicate device for the identification of HSV-2 at Study Sites Combined:

| Table 5.11: Combined Study Sites - Subject Device compared<br>to Predicate Device for the Typing of HSV-2 | | | |
|-----------------------------------------------------------------------------------------------------------|-----------------------------------------------------------|-----------------|----------------|
| Specimen (106 specimens) | Predicate Device HSV-2<br>(Current ELVIS Kit Formulation) | | |
| | Pos | Neg | |
| Subject Device HSV-2<br>(D³ ELVIS) | Pos | 145 | 6 |
| | Neg | 1 | 98 |
| Positive Percent Agreement (PPA) | | 99.3% (145/146) | |
| 95% CI-PPA | | 96.2 – 100% | |
| Negative Percent Agreement (NPA) | | | 94.2% (98/104) |
| 95% CI-NPA | | | 87.9 – 97.9% |

Table 5.12 shows the comparison of the Subject device with the Predicate device for the identification of HSV-2 at Study Sites Combined:

| Table 5.12: Combined Study Sites - Subject Device compared<br>to Predicate Device for the Typing of HSV-1 | | | |
|-----------------------------------------------------------------------------------------------------------|-----------------------------------------------------------|-----|---|
| Specimen (36 specimens) | Predicate Device HSV-1<br>(Current ELVIS Kit Formulation) | | |
| | Pos | Neg | |
| Subject Device HSV-1<br>(D³ ELVIS) | Pos | 90 | 1 |
| | Neg | 0 | 7 |
| Positive Percent Agreement (PPA) | 100% (32/32) | | |

{13}------------------------------------------------

ELVIS®HSV ID & D3 Typing Test System

| 95% CI-PPA | 96.0 – 100% |
|----------------------------------|--------------|
| Negative Percent Agreement (NPA) | 87.5% (7/8) |
| 95% CI-NPA | 47.3 – 99.7% |

The analytical testing and study results from the combined sites demonstrate that the ELVIS®HSV ID and D3 Typing Test System results when compared to the results obtained with the FDA-cleared ELVIS®HSV ID/Typing Test System demonstrated adequate performance to be considered substantially equivalent for the qualitative isolation and identification of HSV-1 and HSV-2 in ELVIS®HSV cell cultures.

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Image /page/14/Picture/0 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features a stylized depiction of an eagle or bird-like figure with three curved lines representing its wings or body. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular pattern around the emblem. The logo is in black and white and appears to be slightly pixelated.

#### DEPARTMENT OF HEALTH & HUMAN SERVICES

Public Health Service

Food and Drug Administration 10903 New Hampshire Avenue Building 66 Silver Spring, MD 20993

### AUG 2 8 2009

Ronald H. Lollar Diagnostic Hybrids, Inc. 1055 East State Street Suite 100 Athens, Ohio 45701

Re: K091753

Trade/Device Name: ELVIS HSV ID and D3 Typing Test System Regulation Number: 21 CFR 866.3305 Regulation Name: Herpes simplex virus serological assays Regulatory Class: Class II Product Code: GQL Dated: June 12, 2009 Received: June 16, 2009

Dear Mr. Lollar:

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

{15}------------------------------------------------

### Page 2 - Mr. Lollar

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 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 vours.

Fally atter
Fally Haiat, M.Sc., Ph.D.

Sally How 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): k091753

# Device Name: ELVIS® HSV ID and D3 Typing Test System

Indications For Use:

The ELVIS®HSV ID and D3 Typing Test System provides Cells, Replacement Medium and Test Reagents for the culture, qualitative identification and typing of herpes simplex virus (HSV) from cutaneous or mucocutaneous specimens as an aid in the diagnosis of HSV type 1 (HSV-1) and HSV type 2 (HSV-2) infections. The performance characteristics of this assay have not been established for antiviral therapy, prenatal monitoring or use with cerebral spinal fluid specimens.

Prescription Use X (Part 21 CFR 801 Subpart D) AND/OR

Over-The-Counter Use (21 CFR 807 Subpart C)

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

Concurrence of CDRH, Office of In Vitro Diagnostic Devices (OIVD)

K.W. ROUTH for US
Division Sign-Off

Jivision Sign

Office of In Vitro Diagnostic Device Office of Safety

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510(k) 091753