The Herpes Group IgG ELISA kit is an Enzyme-Linked Immunosorbent Assay (ELISA) for qualitative determination of IgG antibodies in human serum to Herpes simplex virus. The Herpes Group IgG ELISA kit may be used to evaluate paired sera for the presence seroconversions of IgG as an aid in the diagnosis of Herpes simplex virus infection.

Device Description

The Herpes Group IgG ELISA test is an enzyme linked immunosorbent assay to detect IgG antibodies to Herpes simplex virus. Purified Herpes Group antigen is attached to a solid phase microtiter well. Diluted test sera is added to each well. If the antibodies are present that recognize the antigen, they will bind to the antigen in the well. After incubation the wells are washed to remove unbound antibody. An enzyme labeled anti-human IgG is added to each well. If antibody is present it will bind to the antibody attached to the antigen on the well. After incubation the wells are washed to remove unbound conjugate. A substrate solution is added to each well. If enzyme is present the substrate will undergo a color change. After an incubation period the reaction is stopped and the color intensity is measured photometrically, producing an indirect measurement of specific antibody in the patient specimen.

AI/ML Overview

Here's an analysis of the acceptance criteria and study proving the device meets them, based on the provided text:

Acceptance Criteria and Device Performance

Acceptance Criteria	Reported Device Performance
Relative Sensitivity (compared to Clark HSV 1 & HSV 2 ELISA)	Overall: 98.9% (95% CI: 97.9% - 100%) Site 1: 99.1% Site 2: 97.9% Site 3: 99.1% Site 4: 100.0%
Relative Specificity (compared to Clark HSV 1 & HSV 2 ELISA)	Overall: 96.7% (95% CI: 94.2% - 99.1%) Site 1: 94.7% Site 2: 100% Site 3: 94.7% Site 4: 96.2%
Relative Agreement (compared to Clark HSV 1 & HSV 2 ELISA)	Overall: 98.1% (95% CI: 97.0% - 99.2%) Site 1: 97.2% Site 2: 98.6% Site 3: 97.7% Site 4: 98.9%
Precision (Inter-site, %CV)	Generally <15% CV for most sera Serum 1: 9.21% Serum 2: 12.6% Serum 3: 9.08% Serum 4: 13.5% Serum 5: 15.1% Serum 6: 121% Serum 7: 150%
Seroconversion Sensitivity (compared to CF)	100% for serum meeting paired sera criteria (evaluating 11 pairs)
Cross-Reactivity (with EBV, CMV, VZV IgG antibodies)	No cross-reactivity observed (values for Herpes Group IgG were low, while other antibodies were present)
Agreement with CDC Panel	96.9% total agreement (excluding 2 equivocals) 95.7% agreement with positive specimens 100% agreement with negative specimens

Study Information

Sample sizes used for the test set and the data provenance:
- Total Sample Size (Overall Comparison Study): 603 sera (excluding equivocals from individual calculations).
- Individual Site Sample Sizes:
  - Site 1: 187 sera (181 used in calculations)
  - Site 2: 152 sera (145 used in calculations)
  - Site 3: 176 sera (170 used in calculations)
  - Site 4: 88 sera (88 used in calculations)
- Precision Study: 7 sera, each assayed 10 times at 3 different sites (total 90 assays per serum).
- CF Paired Serum Study: 20 serum pairs, with 11 evaluable pairs.
- Cross-Reactivity Study: 9 serum samples containing IgG antibodies to EBV, CMV, and VZV.
- CDC Panel: Not explicitly stated, but described as containing 72% positive and 28% negative samples.
- Data Provenance:
  - Site 1: R&D laboratory at a commercial company in Maryland. Frozen sera from normals, ages 12-83, various genders, geographical areas. Retrospective.
  - Site 2: R&D laboratory at a commercial company in New York. Frozen sera from normals, ages 17-59, various genders, geographical areas. Retrospective.
  - Site 3: Clinical laboratory in Pennsylvania. Prospective samples sent for Herpes antibody testing. Prospective.
  - Site 4: Clinical laboratory in Wisconsin. Frozen random normal samples. Retrospective.
  - CF Paired Serum Study: From patients suspected of having acute Herpes simplex infection.
  - Cross-Reactivity Study: Serum samples containing specific antibodies.
  - CDC Panel: Serum panel obtained from the CDC.
Number of experts used to establish the ground truth for the test set and the qualifications of those experts:
- The ground truth for the comparison study (relative sensitivity/specificity/agreement) was established by the Clark HSV 1 and HSV 2 ELISA assays (predicate device), not human experts.
- For the CF Paired Serum Study, the ground truth was Complement Fixation (CF), a laboratory method, not human experts.
- For the CDC panel, the ground truth was "masked, characterized serum panel" with established positive and negative samples, but the method of characterization (and thus involvement of experts) is not specified.
Adjudication method (e.g., 2+1, 3+1, none) for the test set:
- There was no stated adjudication method involving human experts for the test set. The comparison was directly against the results of the predicate device (Clark HSV 1 and HSV 2 ELISA).
If a multi reader multi case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance:
- No, a multi-reader multi-case (MRMC) comparative effectiveness study was not done. This study evaluates an ELISA test kit, which is an automated diagnostic assay, and not an AI or human-assisted image interpretation tool. Therefore, the concept of "human readers improve with AI vs without AI assistance" does not apply.
If a standalone (i.e., algorithm only without human-in-the-loop performance) was done:
- Yes, this study represents a standalone performance evaluation of the Wampole Herpes Group IgG ELISA kit. This is an "algorithm only" (or assay-only, in this context) device designed for qualitative determination of IgG antibodies. Its performance is measured directly against a predicate device and other established lab methods, without human interpretation as part of the primary diagnostic step.
The type of ground truth used (expert consensus, pathology, outcomes data, etc.):
- Comparison Studies: The "ground truth" was established by the predicate device, the Clark HSV 1 and HSV 2 ELISA assays. The report explicitly states: "There was not an attempt to correlate the assay's results with disease presence or absence. No judgment can be made on the comparison assay's accuracy to predict disease." This means the ground truth is relative to another diagnostic test, not a definitive clinical diagnosis or pathology.
- CF Paired Serum Study: The ground truth was based on Complement Fixation (CF) results for seroconversion.
- Cross-Reactivity Study: The ground truth was based on known presence/absence of other specific antibodies in the serum.
- CDC Panel: "Masked, characterized serum panel" with established positive and negative results, though specific methodology for characterization is not detailed.
The sample size for the training set:
- The document does not provide information on a training set. For an ELISA kit, which is a biochemical assay rather than a machine learning algorithm, the concept of a "training set" in the AI sense is generally not applicable. Batch validation and standardization would be part of manufacturing, but not in the same way as training an AI model.
How the ground truth for the training set was established:
- Not applicable, as no training set (in the context of AI/machine learning) is described.

Summary

{0}------------------------------------------------

KA63645

FEB - 4 1997

Summary of Safety and Effectiveness Information Herpes Group IgG ELISA Test Kit

I. Immuno Probe Inc. 1306 Bailes Lane, Suite F Frederick, Maryland 21701 Contact person: William Boteler Telephone: 301-695-7920 Date of preparation: Jan 23, 1997

Description of Device II.

III. Predicate Device

The Herpes Group IgG ELISA test is substantially equivalent to Clarks HSV I and HSV II ELISA tests. Equivalence is demonstrated by the following comparative results:

{1}------------------------------------------------

Performance Characteristics

Relative sensitivity and specificity. Four different sites compared the Wampole Herpes Group IgG ELISA test relative to Clarks HSVI and HSVII ELISA assays. The first site was a R&D laboratory at a commercial company located in Maryland. The frozen sera were from normals with ages from 12-83, with various gender, and geographical areas. The results of the study are compiled and summarized in Table 1.

Note: Please be advised the "refers to the comparison of this assay's results to that of a similar assay. There was not an attempt to correlate the assay's results with disease presence or absence. No judgment can be made on the comparison assay's accuracy to predict disease.

Table 1 Comparison of Herpes Group IgG ELISA and Clark HSV 1 and HSV 2 Study 1

Wampole Herpes Group IgG ELISA

	+	eq	-		Total
	+*	104	1	1	106
ClarkHSV 1 &HSV 2	eq**	3	0	0	3
	_***	4	2	72	78
	Total	111	3	73	187

Relative Sensitivity = 104/105 = 99.1%	95% Confidence interval = 97.2% - 100%
Relative Specificity = 72/76 = 94.7%	95% Confidence interval = 89.6% - 99.9%
Relative Agreement = 176/181 = 97.2%	95% Confidence interval = 94.8% - 99.7%

Equivocals were not included in the above calculations. The 95% confidence intervals were calculated using the normal method.

Indicates positive on Clark HSV 1 and/or Clark HSV 2.

** Indicates equivocal on Clark HSV 1 and/or Clark HSV 2.

{2}------------------------------------------------

The second site was a R&D laboratory at a commercial company located in New York. The frozen sera were from normals with ages from 17-59, with various gender, and geographical areas. The results of the study are compiled and summarized in Table 2.

Table 2 Comparison of Herpes Group IgG ELISA and Clark HSV 1 and Clark HSV 2 Study 2

	+	eq	-	Total
+*	92	6	2	100
ClarkHSV 1 andHSV 2	eq**	1	0	0	1
	***	0	0	51	51
	Total	93	6	53	152

Wampole Herpes Group IgG ELISA

Relative Sensitivity = 92/94 = 97.9%	95% Confidence interval = 94.9% - 100%
Relative Specificity = 51/51 = 100%	95% Confidence interval = 94.2% - 100%
Relative Agreement = 143/145 = 98.6%	95% Confidence interval = 96.7% - 100%

Equivocals were not included in the above calculations.

The 95% confidence intervals were calculated using the normal method.

The 95% confidence interval for specificity was calculated assuming one false positive.

Indicates positive on Clark HSV 1 and/or Clark HSV 2.

** Indicates equivocal on Clark HSV 1 and/or Clark HSV 2.

{3}------------------------------------------------

The third site was a clinical laboratory located in Pennsylvania. The sera were prospective samples sent in to the lab for Herpes antibody testing. The results of the studies are compiled and summarized in Table 3.

Table 3 Comparison of Herpes Group IgG ELISA and Clark HSV 1 and Clark HSV 2 Study 3

	+	eq	-	Total
+*	112	0	1	113
ClarkHSV 1 andHSV 2	eq**	1	0	1	2
	-***	3	4	54	61
	Total	116	4	56	176

Wampole Herpes Group IgG ELISA

Relative Sensitivity = 112/113 = 99.1%	95% Confidence interval = 97.4% - 100%
Relative Specificity = 54/57 = 94.7%	95% Confidence interval = 88.8% - 100%
Relative Agreement = 166/170 = 97.7%	95% Confidence interval = 95.3% - 100%

Equivocals were not included in the above calculations. The 95% confidence intervals were calculated using the normal method.

Indicates positive on Clark HSV 1 and/or Clark HSV 2.

** Indicates equivocal on Clark HSV 1 and/or Clark HSV 2.

{4}------------------------------------------------

The forth site was a clinical laboratory located in Wisconsin. The frozen sera were random normal samples. The results of the studies are compiled and summarized in Table 4.

Table 4 Comparison of Herpes Group IgG ELISA and Clark Herpes 1 & 2 Study 4

Wampole Herpes Group IgG ELISA + eg

		+	-	Total
ClarkHerpes 1 & 2	+*	62	0	62
	eq**	0	0	0
	-***	1	25	26
	Total	63	25	88

Relative Sensitivity = 62/62 = 100.0%	95% Confidence interval = 95.3% - 100%
Relative Specificity = 25/26 = 96.2%	95% Confidence interval = 96.2% - 100%
Relative Agreement = 87/88 = 98.9%	95% Confidence interval = 98.9% - 100%

Equivocals were not included in the above calculations.

The 95% confidence intervals were calculated using the normal method. The 95% confidence interval for sensitivity was calculated assuming one false negative.

Indicates positive on Clark HSV 1 and/or Clark HSV 2.

** Indicates equivocal on Clark HSV 1 and/or Clark HSV 2.

{5}------------------------------------------------

The results of the four studies are compiled and summarized in Table 5.

Table 5 Comparison of Herpes Group IgG ELISA and Clark HSV 1 and HSV 2

		+	eq	-	Total
ClarkHSV 1 &HSV 2	+*	370	7	4	381
	eq**	5	0	1	6
	-***	7	6	203	216
	Total	382	13	208	603

Wampole Herpes Group IgG ELISA

Relative Sensitivity = 370/374 = 98.9%	95% Confidence interval = 97.9% - 100%
Relative Specificity = 203/210 = 96.7%	95% Confidence interval = 94.2% - 99.1%
Relative Agreement = 573/584 = 98.1%	95% Confidence interval = 97.0% - 99.2%

Equivocals were not included in the above calculations. The 95% confidence intervals were calculated using the normal method.

Indicates positive on Clark HSV 1 and/or Clark HSV 2.

** Indicates equivocal on Clark HSV 1 and/or Clark HSV 2.

*** Indicates negative on both Clark HSV 1 and Clark HSV 2.

{6}------------------------------------------------

Precision. Seven sera were assayed ten times each on three different assays at three different sites. The intersite precision is shown in Table 6. With appropriate technique the user should obtain precision of <15% CV.

	(n = 90)
Sera #	X	SD	CV
1.	3.81	0.351	9.21%
2.	2.03	0.255	12.6%
3.	3.16	0.287	9.08%
4.	2.01	0.272	13.5%
5.	1.31	0.198	15.1%
6.	0.09	0.109	121%
7.	0.03	0.045	150%

Table 6 Herpes Group IgG ELISA Inter Site Precision Study

CF Paired Serum Study. Twenty serum pairs tested by CF from patients suspected of having acute Herpes simplex infection were assayed on the Herpes Group IgG ELISA assay. Each serum pair was evaluated to determine a seroconversion. Six serum pairs could not be evaluated due to the acute being positive.

Three serum pairs could not be evaluated due to the convalescent being negative. The remaining eleven pairs all demonstrated a seroconversion thus giving a 100% sensitivity versus CF for showing a seroconversion in antibody for serum meeting the paired sera criteria.

{7}------------------------------------------------

Serum containing IgG antibody detectable by ELISA to Epstein Barr 4. Cross-Reactivity. Virus, Cytomegalovirus, and Varicella Zoster Virus were assayed. The data summarized in Table 7 indicates that antibodies to these Herpes Viruses do not cross-react with the Herpes IgG ELISA kit.

SERUM	HerpesGroup IgG	EBV VCA	CMV	VZV
1	0.17	2.6	Negative	2.7
2	0.05	2.3	Negative	1.6
3	0.00	Negative	Negative	2.2
4	0.00	1.8	Negative	2.0
5	0.14	6.3	1.2	2.1
6	0.08	2.4	Negative	3.3
7	0.09	1.1	Negative	2.1
8	0.12	7.2	1.1	3.2
9	0.18	Negative	2.9	3.0

Table 7 Cross-reactivity Study

Sera ≥ 1.10 were considered positive. Sera ≤ 0.90 were considered negative.

The following information is from a serum panel obtained from the CDC and tested by Wampole Herpes Group IgG ELISA. The results are presented as a means to convey further information on the performance of this assay with a masked, characterized serum panel. This does not imply an endorsement of the assay by the CDC.

The panel consists of 72% positive and 28% negative samples. Excluding two equivocals, the Wampole Herpes Group ELISA demonstrated 96.9% total agreement with the CDC results. Of the results obtained by the Wampole Herpes Group IgG ELISA, excluding two equivocals, there was 95.7% agreement with the positive specimens, and 100% agreement with the negative specimens.

Regulation Number and Section

§ 866.3305 Herpes simplex virus serological assays.

(a)
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.(b)
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).