EUROIMMUN ANTI-HSV-2 ELISA (IGG) AND ANTI-HSV-1 ELISA (IGG) KIT

K061239 · Euroimmun Us, LLC · MYF · Jun 28, 2007 · Microbiology

Device Facts

• Record ID: K061239
• Device Name: EUROIMMUN ANTI-HSV-2 ELISA (IGG) AND ANTI-HSV-1 ELISA (IGG) KIT
• Applicant: Euroimmun Us, LLC
• Product Code: MYF · Microbiology
• Decision Date: Jun 28, 2007
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3305
• Device Class: Class 2

Indications for Use

The EUROIMMUN Anti-HSV-1 ELISA (IgG) is intended for the qualitative determination of IgG class antibodies against Herpes simplex virus type 1 (HSV-1) specific glycoprotein C1 in human serum. It is intended for the presumptive diagnosis of type specific HSV-1 infection with EUROIMMUN Anti-HSV-2 EUSA (IgG) in persons suspected of herpes viral infection. The EUROIMMUN Anti-HSV-2 ELISA (IgG) is intended for the qualitative determination of IgG class antibodies against herpes simplex virus type 2 (HSV-2) specific glycoprotein G2 in human serum. It is intended for the presumptive diagnosis of type specific HSV-2 infection with EUROIMMUN Anti-HSV-1 EUSA (lgG) in persons suspected of herpes viral infection.

Device Story

In vitro diagnostic ELISA kits for qualitative detection of IgG antibodies against HSV-1 and HSV-2; utilizes human serum samples; detects specific viral glycoproteins (C1 for HSV-1, G2 for HSV-2); intended for use in clinical laboratory settings by trained personnel; results aid in presumptive diagnosis of type-specific HSV infection; supports clinical decision-making regarding patient management for suspected herpes viral infections.

Clinical Evidence

Clinical performance evaluated across multiple studies (n=397 for HSV-1; n=421 for HSV-2) using Western blot as reference. HSV-1 sensitivity ranged 98.1-100%; specificity 71.4-100%. HSV-2 sensitivity ranged 94.5-100%; specificity 76.9-100%. Comparison to predicate kits showed high agreement (HSV-1: 96.5% negative, 98.3% positive; HSV-2: 97.4% negative, 93.9% positive). Type specificity confirmed at 97.6% (HSV-1) and 97.0% (HSV-2).

Technological Characteristics

ELISA-based immunoassay. Microtiter wells coated with affinity-purified HSV-1 glycoprotein C1 or HSV-2 glycoprotein G2. Detection via rabbit anti-human IgG HRP conjugate and TMB substrate. Qualitative output. Manual or automated plate processing.

Indications for Use

Indicated for qualitative detection of IgG antibodies against HSV-1 (glycoprotein C1) or HSV-2 (glycoprotein G2) in human serum. Used for presumptive diagnosis of type-specific HSV infection in patients suspected of herpes viral infection.

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).

Submission Summary (Full Text)

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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY

A. 510(k) Number:
K061239

B. Purpose for Submission:
New device(s)

C. Measurand:
Anti-HSV-1 IgG antibodies and Anti-HSV-2 IgG antibodies

D. Type of Test:
ELISA

E. Applicant:
EUROIMMUN US LLC

F. Proprietary and Established Names:
EUROIMMUN Anti-HSV-1 ELISA (IgG) Kit and EUROIMMUN Anti-HSV-2 ELISA (IgG) Kit

G. Regulatory Information:

1. Regulation section: 21CFR 866.3305: Herpes Simplex Virus Serological Reagents
2. Classification: Class: II
3. Product code:
MXJ: Enzyme linked immunosorbent assay, herpes simplex virus, hsv-1
MYF: Enzyme linked immunosorbent assay, herpes simplex virus, hsv-2
4. Panel: 83 Microbiology

H. Intended Use:
HSV-1 Assay: The EUROIMMUN Anti-HSV-1 ELISA (IgG) is intended for the qualitative determination of IgG class antibodies against Herpes simplex virus type 1 (HSV-1) specific glycoprotein C1 in human serum. It is intended for the

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presumptive diagnosis of type specific HSV-1 infection in conjunction with EUROIMMUN Anti-HSV-2 ELISA (IgG) in persons suspected of herpes viral infection.

HSV-2 Assay: The EUROIMMUN Anti-HSV-2 ELISA (IgG) is intended for the qualitative determination of IgG class antibodies against Herpes simplex virus type 2 (HSV-2) specific glycoprotein G2 in human serum. It is intended for the presumptive diagnosis of type specific HSV-2 infection in conjunction with EUROIMMUN Anti-HSV-1 ELISA (IgG) in persons suspected of herpes viral infection.

2. Indication(s) for use:

HSV-1 Assay: The EUROIMMUN Anti-HSV-1 ELISA (IgG) is intended for the qualitative determination of IgG class antibodies against Herpes simplex virus type 1 (HSV-1) specific glycoprotein C1 in human serum. It is intended for the presumptive diagnosis of type specific HSV-1 infection in conjunction with EUROIMMUN Anti-HSV-2 ELISA (IgG) in persons suspected of herpes viral infection.

HSV-2 Assay: The EUROIMMUN Anti-HSV-2 ELISA (IgG) is intended for the qualitative determination of IgG class antibodies against Herpes simplex virus type 2 (HSV-2) specific glycoprotein G2 in human serum. It is intended for the presumptive diagnosis of type specific HSV-2 infection in conjunction with EUROIMMUN Anti-HSV-1 ELISA (IgG) in persons suspected of herpes viral infection.

3. Special conditions for use statement(s):

For prescription use

4. Special instrument requirements:

None

I. Device Description:

HSV-1 Assay: The test kit contains microtiter wells coated with affinity purified glycoprotein C1 isolated from HSV-1. In the first reaction step, diluted patient samples, calibrators and controls are incubated in the wells. Anti-HSV-1 antibodies will bind to the antigens coated in the microtiter wells. The wells are washed to remove any unbound proteins and non-specific antibodies. In a second reaction step, rabbit anti-human IgG HRP enzyme conjugate is added to each well. The enzyme conjugate will bind to any wells that have human IgG binding to the HSV-1 antigen. The wells are washed to remove any unbound HRP enzyme conjugate. 3,3,5,5 tetramethylbenzidine (TMB) enzyme substrate is added. If the HRP enzyme is present in the well (positive reaction), the HRP enzyme will react with the TMB substrate and produce a blue color.

HSV-2 Assay: The test kit contains microtiter wells coated with affinity purified

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glycoprotein G2 isolated from HSV-2. In the first reaction step, diluted patient samples, calibrators and controls are incubated in the wells. Anti-HSV-2 antibodies will bind to the antigens coated in the microtiter wells. The wells are washed to remove any unbound proteins and non-specific antibodies. In a second reaction step, rabbit anti-human IgG HRP enzyme conjugate is added to each well. The enzyme conjugate will bind to any wells that have human IgG binding to the HSV-2 antigen. The wells are washed to remove any unbound HRP enzyme conjugate. 3,3,5,5 tetramethylbenzidine (TMB) enzyme substrate is added. If the HRP enzyme is present in the well (positive reaction), the HRP enzyme will react with the TMB substrate and produce a blue color.

## J. Substantial Equivalence Information:

1. Predicate device name(s):
HerpeSelect® 1 ELISA IgG
HerpeSelect® 2 ELISA IgG

2. Predicate 510(k) number(s):
K021429 (HSV-1)
K021486 (HSV-2)

3. Comparison with predicate:

| Similarities | | |
| --- | --- | --- |
| Item | Device | Predicate |
| HSV-1 Assay | | |
| Specimen Type | Human serum | Human serum |
| Method and type | Qualitative | Qualitative |
| | ELISA | ELISA |
| HSV-2 Assay | | |
| Method and type | Qualitative | Qualitative |
| Specimen Type | Human serum | Human serum |
| Antigen used | Glycoprotein G-2 | Glycoprotein G-2 |
| Differences | | |
| Item | Device | Predicate |
| HSV-1 Assay | | |
| Antigen used | Glycoprotein C-1 | Glycoprotein G-1 |

## K. Standard/Guidance Document Referenced (if applicable):

Not applicable

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L. Test Principle:

Enzyme linked immunosorbent assay, ELISA

M. Performance Characteristics (if/when applicable):

1. Analytical performance:

a. Precision/Reproducibility:

HSV-1 Assay: The reproducibility of the HSV-1 test was investigated by determining the intra- and inter-assay coefficients of variation using 3 sera. The intra-assay CVs are based on 20 determinations and the inter-assay CVs on 4 determinations performed in 4-6 different test runs. The inter-assay CV ranged from 2.5 to 7.2% for positive specimens.

HSV-2 Assay: The reproducibility of the HSV-2 test was investigated by determining the intra- and inter-assay coefficients of variation using 4 to 6 different sera. The intra-assay CVs are based on 20 determinations and the inter-assay CVs on 4 determinations performed in 4 different test runs. The inter-assay CV ranged from 2.3 to 8.4% for positive specimens.

b. Linearity/assay reportable range:

Not applicable

c. Traceability, Stability, Expected values (controls, calibrators, or methods):

Not applicable

d. Detection limit:

Not applicable

e. Analytical specificity:

HSV-1 Assay: This ELISA showed no serological cross reactivity with sera positive for the following: EBV-CA (n = 12), CMV (n = 6), VZV (n = 12), Adenovirus (n = 12), RSV (n = 12), Parainfluenza types 1-4 (n = 12), Influenza A (n = 12), Influenza B (n = 12), Mycoplasma pneumoniae (n = 8), Mumps (n = 12), Measles (n = 12), Rubella (n = 12), Chlamydia pneumoniae (n = 4), Helicobacter pylori (n = 7).

HSV-2 Assay: This ELISA showed no serological cross reactivity with sera positive for the following: HSV-1 (n = 12); EBV-CA (n = 12); CMV (n = 12); VZV (n = 12); Adenovirus (n = 12); RSV (n = 12); Parainfluenza types 1-4 (n = 12); Influenza A (n = 12); Influenza B (n = 12); Mycoplasma pneumoniae (n = 12); Mumps (n = 12); Measles (n = 12); Rubella (n = 12); Toxoplasma (n = 12); Chlamydia pneumoniae (n = 12); Helicobacter pylori (n = 12).

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f. Assay cut-off:

Not applicable

2. Comparison studies:

a. Method comparison with predicate device:

HSV-1 Assay

Sensitivity and specificity: In 4 clinical studies, 397 samples were tested with the EUROIMMUN Anti-HSV-1 ELISA (IgG) and Western blot as the reference method.

Study 1: Hundred (100) prospective samples from the US were tested with the EUROIMMUN Anti-HSV-1 ELISA (IgG) and Western blot as the reference method. This group consisted of 38 men and 57 women (and 5 unknown) with an average age of 36 years (range: 19-84 years).

Specificity = 35 / 35 = 100 %

Sensitivity = 65 / 65 = 100 %

Study 2: Two-hundred-fifty-four (254) samples, consisting of 186 samples from a risk group and 68 routine samples (both Muenster, Germany) were tested with the EUROIMMUN Anti-HSV-1 ELISA (IgG) and Western blot as the reference method.

Specificity = 62 / 62 = 100 %

Sensitivity = 187 / 187 = 100 %

Study 3: Twenty-five (25) member performance panel obtained commercially from Boston Biomedica Inc. were tested with the EUROIMMUN Anti-HSV-1 ELISA (IgG) and Western blot as the reference method.

Specificity = 5 / 7 = 71.4 %

Sensitivity = 14 / 14 = 100 %

Study 4: Eighteen (18) characterized samples obtained from INSTAND (Institute for Standardization and Documentation in Medical Laboratory) were tested with the EUROIMMUN Anti-HSV-1 ELISA (IgG) and Western blot as the reference method.

Specificity = 5 / 5 = 100 %

Sensitivity = 13 / 13 = 100 %

CDC panel: A panel of hundred (100) characterized samples obtained from the CDC were tested with the EUROIMMUN Anti-HSV-1 ELISA (IgG).

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Negative Agreement = 35 / 45 = 77.8 %
Positive Agreement = 51 / 52 = 98.1 %

INSTAND samples: 28 clinically characterized patient samples (Inter-laboratory test samples of the INSTAND, Germany) were examined with the EUROIMMUN Anti-HSV-1 ELISA (IgG). The test shows an agreement of 100%.

Negative Agreement = 8 / 8 = 100 %
Positive Agreement = 20 / 20 = 100 %

Comparison to predicate kit: A study was conducted at a hospital clinical laboratory comparing the performance of the EUROIMMUN Anti-HSV-1 IgG ELISA and a kit in current distribution. 259 prospective samples from the US were tested.

Negative Agreement = 83 / 86 = 96.5 %
Positive Agreement = 170 / 173 = 98.3 %

Type specificity: Type specificity was confirmed using sera of patients serologically positive for Anti-HSV-1 IgG and negative for Anti-HSV-2 IgG and vice versa. 168 samples were tested with the EUROIMMUN Anti-HSV-1 ELISA and with another FDA-cleared ELISA as reference method. A type specificity of 97.6% was observed.

Negative Agreement = 30 / 31 = 96.8 %
Positive Agreement = 134 / 137 = 97.8 %

## HSV-2 Assay

Sensitivity and specificity: In 5 clinical studies, 421 samples were tested with the EUROIMMUN Anti-HSV-2 ELISA (IgG) and Western blot as the reference method.

Study 1: Hundred (100) prospective samples from the US were tested with the EUROIMMUN Anti-HSV-1 ELISA (IgG) and Western blot as the reference method. This group consisted of 38 men and 57 women (and 5 unknown) with an average age of 36 years (range: 19-84 years).

Specificity = 64 / 64 = 100 %
Sensitivity = 36 / 36 = 100 %

Study 2: Two-hundred-fifty-four (254) samples, consisting of 186 samples from a risk group and 68 routine samples (both Muenster, Germany) were tested with the EUROIMMUN Anti-HSV-2 ELISA (IgG) and Western blot as the reference method.

Specificity = 148 / 160 = 92.5 %
Sensitivity = 87 / 87 = 100 %

Study 3: Twenty-five (25) member performance panel obtained commercially from

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Boston Biomedica Inc. were tested with the EUROIMMUN Anti-HSV-2 ELISA (IgG) and Western blot as the reference method.

$$
\begin{array}{l}
\text{Specificity} = 10 / 13 = 76.9\% \\
\text{Sensitivity} = 9 / 9 = 100\% \\
\end{array}
$$

Study 4: Eighteen (18) characterized samples obtained from INSTAND (Institute for Standardization and Documentation in Medical Laboratory) were tested with the EUROIMMUN Anti-HSV-2 ELISA (IgG) and Western blot as the reference method.

$$
\begin{array}{l}
\text{Specificity} = \mathrm{N}/\mathrm{A} \\
\text{Sensitivity} = 17 / 18 = 94.5\% \\
\end{array}
$$

Study 5: Twenty-four (24) stored samples (Chennai, India) were tested with the EUROIMMUN Anti-HSV-2 ELISA (IgG) and Western blot as the reference method.

$$
\begin{array}{l}
\text{Specificity Agreement} = 20 / 20 = 100\% \\
\text{Sensitivity} = 4 / 4 = 100\% \\
\end{array}
$$

CDC panel: A panel of hundred (100) characterized samples obtained from the CDC were tested with the EUROIMMUN Anti-HSV-2 ELISA (IgG).

$$
\begin{array}{l}
\text{Negative Agreement} = 44 / 50 = 88.0\% \\
\text{Positive Agreement} = 49 / 50 = 98.0\% \\
\end{array}
$$

Comparison to predicate kit: A study was conducted at a hospital clinical laboratory comparing the performance of the EUROIMMUN Anti-HSV-2 IgG ELISA and a kit in current distribution. 259 prospective samples from the US were tested.

$$
\begin{array}{l}
\text{Negative Agreement} = 188 / 193 = 97.4\% \\
\text{Positive Agreement} = 62 / 66 = 93.9\% \\
\end{array}
$$

Type specificity: Type specificity was confirmed using sera of patients serologically positive for Anti-HSV-1 IgG and negative for Anti-HSV-2 IgG and vice versa. 168 samples were tested with the EUROIMMUN Anti-HSV-2 ELISA and with another FDA-cleared ELISA as reference method. A type specificity of 97.0% was observed.

$$
\begin{array}{l}
\text{Negative Agreement} = 133 / 137 = 97.1\% \\
\text{Positive Agreement} = 30 / 31 = 96.8\% \\
\end{array}
$$

b. Matrix comparison:

Not applicable

3. Clinical studies:

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a. Clinical Sensitivity:
Not applicable

b. Clinical specificity:
Not applicable

c. Other clinical supportive data (when a. and b. are not applicable):
Not applicable

4. Clinical cut-off:
Not applicable

5. Expected values/Reference range:
Not applicable

N. Proposed Labeling:
The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.

O. Conclusion:
The submitted information in this premarket notification is complete and supports a substantial equivalence decision.