The Epstein Barr Nuclear Antigen 1 (EBNA-1) IgM kit is an Enzyme-Linked Immunosorbent Assay (ELISA) for the qualitative determination of IgM antibodies in human serum to EBNA-1 antigen. The Clark anti-EBNA-1 IgM assay may be used in conjunction with other Epstein-Barr serologies (VCA IgG, VCA IgM, EA-D IgG, EBNA-1 IgG and heterophile) as an aid in the diagnosis of infectious mononucleosis in adult populations.

Device Description

The Wampole Epstein-Barr Virus Nuclear Antigen 1 (EBNA-1) IgM kit is an Enzyme-Linked Immunosorbent Assays (ELISA) for the qualitative determination of IgM antibodies in human serum to EBNA-1 antigen. The Wampole anti-EBNA-1 IgM assay may be used in conjunction with other Epstein-Barr serologies (VCA IgG, VCA IgM, EA-D IgG, EBNA-1 IgG and heterophile) as an aid in the diagnosis of infectious mononucleosis.

For In Vitro Diagnostic Use Only.

The EBNA-1 IgM ELISA test is an enzyme linked immunosorbent assay to detect IgM antibodies to Epstein-Barr Nuclear Antigen -1. Recombinant EBNA-1 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 IgM 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 provided information regarding the EBNA-1 IgM ELISA Test Kit, presented with the requested structure:

1. Table of Acceptance Criteria and Reported Device Performance

The acceptance criteria are not explicitly stated in the provided text. However, based on the presented performance characteristics, we can infer the implied targets for sensitivity and specificity. The study demonstrates performance against these inferred criteria.

Performance Metric	Implied Acceptance Criteria (Inferred)	Reported Device Performance
Sensitivity	High for acute cases (target > ~50%)	51.4% (95% CI: 34.9%-67.8%)
Specificity (Seronegative)	High (target = 100%)	100% (95% CI: 89.4%-100%)
Specificity (Seropositive)	High (target > ~90%)	94.5% (95% CI: 89.7%-99.3%)
Relative Agreement	High (target > ~80%)	85.3% (95% CI: 79.6%-90.9%)
Precision (Inter-Site CV)	Low variability (e.g., < 15-20% for most samples)	Variable (e.g., 8.60% to 26.36%, with HPC 4.45%, CAL 6.48%, NC 181.66%)
Cross-Reactivity	No significant cross-reactivity with common interfering antibodies (e.g., RF, VZV)	No cross-reactivity with RF or VZV. Some cross-reactivity with HSV and CMV.

2. Sample Size Used for the Test Set and Data Provenance

Test Set Sample Size: 166 selected serum samples
Data Provenance: The document states, "One hundred and sixty six selected serum were tested at a clinical lab." The country of origin is not explicitly stated, but given the company's US presence and the FDA submission, it is likely US-based. The data is retrospective, as the samples were "selected" and characterized prior to testing with the Wampole EBNA-1 IgM kit.

3. Number of Experts Used to Establish the Ground Truth for the Test Set and the Qualifications of Those Experts

The ground truth was established by "characteriz[ing]" the serum samples as seronegative, acute, or seropositive based on other established EBV serologies (VCA IgM, VCA IgG, EA-D IgG, EBNA-1 IgG, and heterophile). The text does not specify the number of experts or their qualifications for this characterization. It implies standard laboratory methods were used.

4. Adjudication Method for the Test Set

No specific adjudication method is mentioned. The ground truth (characterization of samples) appears to be based on results from other established serological tests, implying a consensus based on these diagnostic markers rather than a panel of experts adjudicating individual cases. Equivocal results from the Wampole assay were "not included in the calculations" and were simply "reported as equivocal," indicating no adjudication for these specific outcomes.

5. 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

This is an ELISA (Enzyme-Linked Immunosorbent Assay) test kit, not an AI-powered diagnostic device. Therefore, an MRMC comparative effectiveness study involving human readers and AI assistance is not applicable and was not performed.

6. If a Standalone (i.e. algorithm only, without human-in-the-loop performance) Was Done

Yes, the performance characteristics described (Sensitivity and Specificity, Precision, Cross-Reactivity) represent a standalone evaluation of the EBNA-1 IgM ELISA kit. It is an in vitro diagnostic device that produces a quantitative (ISR Value) or qualitative (Positive/Negative/Equivocal) result based on biochemical reactions, without a human-in-the-loop for interpretation beyond reading the instrument's output.

7. The Type of Ground Truth Used

The ground truth used was based on expert consensus of established serological markers. Samples were categorized as "seronegative," "acute," or "seropositive" using a panel of other EBV serologies: VCA IgM, VCA IgG, EA-D IgG, EBNA-1 IgG, and heterophile anti-body status. This combines multiple diagnostic test results to define the true disease status for comparison.

8. The Sample Size for the Training Set

The document does not explicitly mention a "training set" for the ELISA kit. ELISA kits are typically developed and validated using biochemical principles and reagents. While there would have been internal R&D and analytical validation experiments, the clinical "Performance Characteristics" section focuses on testing against known samples, which serves as the validation or test set.

9. How the Ground Truth for the Training Set Was Established

As no specific training set for a machine learning algorithm is discussed, this question is not applicable to an ELISA test kit. The "ground truth" for the overall development and validation of the assay would stem from the known characteristics of the antigens, antibodies, and patient samples used in its development and analytical testing.

Summary

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

K980598

Summary of Safety and Effectiveness Information EBNA-1 IgM ELISA Test Kit

1 1998 MAY

I. Trinity Biotech US PO Box 1059 Jamestown, NY 14702-1059 Contact person: Ron Cruver Telephone: 716-483-3851 Date of preparation: Feb 11,1998
II. Description of Device

For In Vitro Diagnostic Use Only.

III. Predicate Device

The EBNA-1 IgM ELISA test is substantially equivalent to EBV serology. Equivalence is demonstrated by the following comparative results:

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

Performance Characteristics

1. Sensitivity and Specificity Based on Serum Characterization

One hundred and sixty six selected serum were tested at a clinical lab. The serum from the study were characterized as seronegative ( no serological evidence of past or present EBV infection), acute (VCA IgM and heterophile antibody present, EBNA IgG absent), or seropositive (presence of VCA IgG antibodies and EBNA IgG, no evidence of VCA IgM or heterophile antibody, indicative of past infection). The sensitivity, specificity and agreement of the assay was determined based on this characterization. It was assumed that the EBNA-1 IgM response should be negative for seronegative, and convalescent serum, and positive for acute serum. The results are summarized in Table 1.

Table 1

		AcuteVCA IgM+EBNA IgG -Heterophile +	SeropositiveVCA IgG+EBNA IgG+VCA IgM-Heterophile -	SeronegativeVCA IgG-EBNA IgG -VCA IgM -Heterophile -
WampoleEBNA-1 IgM Equivocal	Positive	19	5	0
		2	8	0
	Negative	18	86	28
	Total	39	99	28

Relative Sensitivity (Acute)	= 19/37 = 51.4%	95% Confidence Interval = 34.9%-67.8%
Relative Specificity (Seronegative)	= 28/28 = 100%	95% Confidence Interval = 89.4%-100%
Relative Specificity (Seropositive)	= 86/91 = 94.5%	95% Confidence Interval = 89.7%-99.3%
Relative Agreement	= 133/156 = 85.3%	95% Confidence Interval = 79.6%-90.9%

Equivocal results were not included in the calculations.

Equivocal results were not retested. They were reported as equivocal.

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

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

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

Precision. The Wampole EBNA-1 IgM EIA was evaluated for precision by testing six sera six times each at site one and ten times each at the second site on three different days. The results are summarized in the table below.

Inter Site Precision Data

Inter Site Precision (n=48)
Serum#	X	S.D.	C.V.
1	1.87	0.161	8.60%
2	1.56	0.166	10.63%
3	2.38	0.156	6.57%
4	1.30	0.132	10.18%
5	0.47	0.050	10.52%
6	0.19	0.051	26.36%
HPC*	3.28	0.146	4.45%
CAL**	2.22	0.144	6.48%
NC*	0.01	0.012	181.66%

X = Mean ISR Value S.D. = Standard Deviation C.V. = Coefficient of Variation

HPC and NC - n=6 ** CAL - n=18

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

Cross-Reactivity. Sera containing IgM antibody detectable by ELISA to Herpes Simplex Virus I & II, Cytomegalovirus, and Varicella Zoster Virus were assayed. Sera containing rhuematoid factor (RF) were also assayed. The data summarized in Table 3 indicates that antibodies to Varicella and sera containing RF do not cross-react with the EBNA-1 IgM EIA kit. There is some cross reactivity to Herpes Simplex Virus and to Cytomegalovirus.

Specificity	EBNA-1 IgM	Alternate Assay
RF +	0.15	1.87	+
RF +	0.09	1.82	+
RF +	0.01	1.73	+
RF +	0.10	1.80	+
RF +	0.10	1.85	+
VZV M +	0.44	3.28	+
VZV M +	0.17	5.46	+
VZV M +	0.10	4.98	+
VZV M +	0.29	2.34	+
VZV M +	0.20	2.18	+
HSV 1 M +	2.53	2.53	+
HSV 1 M +	0.27	1.65	+
HSV 1 M +	0.05	1.34	+
HSV 1 M +	0.08	1.32	+
HSV 2 M +	0.53	1.76	+
HSV 2 M +	0.08	1.60	+
HSV 2 M +	0.03	2.09	+
HSV 2 M +	0.62	1.96	+
CMV M +	1.57	1.23	+
CMV M +	0.98	1.92	+
CMV M +	2.00	3.83	+
CMV M +	0.38	1.32	+

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

Image /page/4/Picture/1 description: The image shows the logo for the Department of Health & Human Services (HHS). The logo features a stylized eagle with its head turned to the left and its wings forming three curved lines. The words "DEPARTMENT OF HEALTH & HUMAN SERVICES • USA" are arranged in a circular pattern around the eagle.

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

1 1998 MAY

Clark Laboratories, Inc. c/o William L. Boteler, Jr. Immuno Probe, Inc. 1306 Bailes Lane, Suite F Frederick, Maryland 21701

K980598 Re: Trade Name: EBNA-1 IgM ELISA Regulatory Class: I Product Code: LLM Dated: February 13, 1998 Received: February 17, 1998

Dear Mr. Boteler:

We have reviewed your Section 510(k) notification of intent to market the device referenced above and we 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). 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 either class II (Special Controls) or class III (Premarket Approval), it may be subject to such additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations. Title 21. Parts 800 to 895. A substantially equivalent determination assumes compliance with the Current Good Manufacturing Practice requirements, as set forth in the Quality System Regulation (OS) for Medical Devices: General regulation (21 CFR Part 820) and that, through periodic OS inspections, the Food and Drug Administration (FDA) will verify such assumptions. Failure to comply with the GMP regulation may result in regulatory action. In addition, FDA may publish further announcements concerning your device in the Federal Register. Please note: this response to your premarket notification submission does not affect any obligation you might have under sections 531 through 542 of the Act for devices under the Electronic Product Radiation Control provisions, or other Federal laws or regulations.

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

Page 2

Under the Clinical Laboratory Improvement Amendments of 1988 (CLIA-88), this device may require a CLIA complexity categorization. To determine if it does, you should contact the Centers for Disease Control and Prevention (CDC) at (770)488-7655.

This letter will allow you to begin marketing your device as described in your 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 Part 801 and additionally 809.10 for in vitro diagnostic devices), please contact the Office of Compliance at (301) 594-4588. Additionally, for questions on the promotion and advertising of your device, please contact the Office of Compliance at (301) 594-4639. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 807.97). Other general information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers Assistance at its toll free number (800) 638-2041 or at (301) 443-6597 or at its internet address "http://www.fda.gov/cdrh/dsmamain.html"

Sincerely yours,

Steven Butman

Steven I. Gutman, M.D., M.B.A. Director Division of Clinical Laboratory Devices Office of Device Evaluation Center for Devices and Radiological Health

Enclosure

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

Page 1 of 1

510(k) Number. Not Known

Device Name: EBNA-1 IgM ELISA

Indications For Use: The Epstein Barr Nuclear Antigen 1 (EBNA-1) IgM kit is an Enzyme-Linked Immunosorbent Assay (ELISA) for the qualitative determination of IgM antibodies in human serum to EBNA-1 antigen. The Clark anti-EBNA-1 IgM assay may be used in conjunction with other Epstein-Barr serologies (VCA IgG, VCA IgM, EA-D IgG, EBNA-1 IgG and heterophile) as an aid in the diagnosis of infectious mononucleosis in adult populations.

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

Concurrence of CDRH, Office of Device Evaluation (ODE)

Prescription Use (Per 21 CFR 801.109)

Over-The Counter Use (Optional Format 1-2-96) --

Alo H

(Division Sign-Off) Division of Clinical Laboratory 510(k) Number_

Regulation Number and Section

§ 866.3235 Epstein-Barr virus serological reagents.

(a)
Identification. Epstein-Barr virus serological reagents are devices that consist of antigens and antisera used in serological tests to identify antibodies to Epstein-Barr virus in serum. The identification aids in the diagnosis of Epstein-Barr virus infections and provides epidemiological information on diseases caused by these viruses. Epstein-Barr viruses are thought to cause infectious mononucleosis and have been associated with Burkitt's lymphoma (a tumor of the jaw in African children and young adults) and postnasal carcinoma (cancer).(b)
Classification. Class I (general controls).