For the qualitative determination of IgM antibodies in human serum to Epstein Barr (recombinant) Viral Capsid antigen (EBV-VCA) antigen. The EBV-VCA IgM assay should be used in conjunction with other Epstein-Barr serologies (EBV-VCA IgG, EBNA-1 IgG, EA-D IgG, EA-D IgM, EBNA-1 IgM and heterophile) as an aid in the diagnosis of infectious monomucleosis. The test can be performed either manually or in conjunction with the MAGO PLUS ™ Automated EIA Processor.

Device Description

The & EBV-VCA IgM ELISA Kit is an enzyme-linked immunosorbent assay (ELISA) for the detection of IgM antibodies to Epstein-Barr Viral Capsid antigen in human serum.

AI/ML Overview

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

Acceptance Criteria and Device Performance

Device Name: EBV-VCA IgM ELISA Kit (also referred to as Is-EBV-VCA IgM Test Kit and 7 VCA IgM ELISA)

Acceptance Criteria Category	Specific Metric	Acceptance Criteria (Implied)	Reported Device Performance
Clinical Performance	Relative Specificity (Convalescent)	High specificity (specific threshold not explicitly stated but implied by high performance)	97.0% (95% CI: 91.6-99.4)
	Relative Sensitivity (Current Infection)	High sensitivity (specific threshold not explicitly stated but implied by high performance)	95.0% (95% CI: 83.1-99.4)
	Relative Specificity (Seronegative)	High specificity (specific threshold not explicitly stated but implied by high performance)	100% (95% CI: 89.1-100)
	Overall Agreement (with predicate/characterized sera)	High agreement (specific threshold not explicitly stated but implied by high performance)	97.1% (95% CI: 93.4-99.1)
Precision	Intra-assay Precision (CV%)	Low variability (threshold not explicitly stated)	Site #1: Positive sera CV% = 6.07-9.92; Negative sera CV% = 8.51-49.37 (higher for negatives, especially low index ones)Site #2: Positive sera CV% = 3.95-6.35; Negative sera CV% = 7.41-17.42Site #3: Positive sera CV% = 4.15-6.22; Negative sera CV% = 7.16-12.82MAGO Plus: Positive sera CV% = 3.53-9.27; Negative sera CV% = 0.00-22.59 (0.00 for one low negative, likely due to rounding or being below detection)
	Interassay Precision (CV%)	Low variability (threshold not explicitly stated)	Site #1: Positive sera CV% = 12.05-13.69; Negative sera CV% = 22.75-39.31Site #2: Positive sera CV% = 6.53-7.34; Negative sera CV% = 9.38-14.80Site #3: Positive sera CV% = 5.08-5.80; Negative sera CV% = 12.70-13.26MAGO Plus: Positive sera CV% = 4.55-8.64; Negative sera CV% = 17.76-0.00 (0.00 for one low negative, likely due to rounding or being below detection)
	Inter-Site Precision (CV%)	Low variability (threshold not explicitly stated)	Positive sera CV% = 11.13-13.43; Negative sera CV% = 20.69-28.77 (NC has 60.11%, likely due to very low index values making CV% highly sensitive)
Cross-Reactivity	Specificity with Potentially Cross-Reactive Sera	Minimal cross-reactivity expected	Some cross-reactivity is suggested: 1/4 anti-VZV IgM positive sera were reactive for anti-VCA IgM; 2/5 anti-CMV IgM positive sera were reactive for anti-VCA IgM. 4/4 anti-HSV positive sera were non-reactive. Results indicate that some specific cross-reactivity should be expected with VZV and CMV.
Method Correlation	Manual and MAGO Plus Results Correlation	Strong correlation (specific threshold not explicitly stated but implied by high performance)	Pearson Correlation Coefficient = 0.990 (indicating strong positive correlation)

Study Details Proving Acceptance Criteria

Sample Size used for the test set and the data provenance:
- Clinical Sensitivity and Specificity: 176 frozen retrospective sera. The provenance (country of origin) is not explicitly stated, but it's referred to as "sera from one hundred and seventy-six patients" which suggests human samples. The study explicitly states it was a "pre-selected, retrospective, population."
- Precision:
  - Intra-assay and Interassay Precision: Four positive and two negative sera (6 unique sera) were assayed 10 times each, in 3 different runs. This was done at 3 different sites.
  - Inter-Site Precision: The document mentions n=90 for serum samples A-F (presumably 90 total measurements across sites for each serum type, or 3 sites * 3 runs * 10 replicates = 90 total for each serum type), and n=36 for CAL (Calibrator), n=18 for LPC (Low Positive Control), n=18 for NC (Negative Control).
- Specificity with Potentially Cross-Reactive Sera: 13 sera.
- Correlation of Manual and MAGO Plus Results: 120 serum samples.
- MAGO Plus Precision: Six sera were assayed 10 times each in 3 different runs (similar to the manual precision study).
Number of experts used to establish the ground truth for the test set and the qualifications of those experts:
- For the "Clinical Sensitivity and Specificity Using Characterized Sera" section, the sera were "characterized using commercially available kits for VCA IgM, VCA IgG, EBNA IgG and heterophile antibodies." This implies an established laboratory testing method rather than human expert opinion directly establishing ground truth for individual cases. No human experts are explicitly mentioned for ground truth establishment.
Adjudication method for the test set:
- No adjudication method (e.g., 2+1, 3+1) is mentioned. The ground truth was established by comparing to results from "commercially available kits" for other EBV serologies.
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 not an MRMC study. The device is an ELISA kit, which is an in vitro diagnostic (IVD) test, not an AI-assisted diagnostic tool for human readers. There is no mention of human readers or AI assistance in this context. The comparison is between the new EBV-VCA IgM ELISA Kit and existing commercial kits/methods.
If a standalone (i.e. algorithm only without human-in-the-loop performance) was done:
- Yes, for the EBV-VCA IgM ELISA Kit, the performance data provided (sensitivity, specificity, precision, cross-reactivity) represents the standalone performance of the diagnostic assay. This assay is a laboratory test, and its results are read and interpreted, but the performance characteristics apply to the kit itself.
- The "Correlation of Manual and MAGO Plus Results" section compares the standalone performance of the kit when processed manually versus using an automated EIA processor (MAGO Plus). Both are "standalone" in the sense of the chemical assay's performance.
The type of ground truth used:
- Clinical Performance: "EBV Serological Status" derived from other "commercially available kits for VCA IgM, VCA IgG, EBNA IgG and heterophile antibodies." This is a lab result-based ground truth (or predicate method comparison) rather than pathology, clinical outcome, or expert consensus on a subjective measure. The document explicitly states: "There was not an attempt to correlate the assay's results with disease presence or absence."
- Precision: Ground truth for positive/negative samples was pre-established (e.g., "four positive and two negative sera").
- Cross-Reactivity: Sera "reactive for IgM antibodies to varicella zoster, cytomegalovirus and herpes simplex virus by EIA". This is again a lab result-based ground truth to check for specific antibody interference.
- Correlation: Comparison between the manual and MAGO Plus results on the same samples.
The sample size for the training set:
- The document does not explicitly describe a "training set" in the context of machine learning or AI. This is an in vitro diagnostic (IVD) assay. The development of such assays typically involves research and development where reagents and protocols are optimized. The "Characterized Sera" mentioned in the clinical performance section serves as the test set for the final validation of the device.
How the ground truth for the training set was established:
- As no "training set" in the AI sense is explicitly mentioned, this question is not fully applicable. For the development and optimization of the ELISA kit itself, ground truth for sample characterization would have been established through well-characterized reference materials, clinical samples with known serological status using predicate methods, or samples from a disease registry. The "Characterized Sera" in the performance study were characterized using "commercially available kits for VCA IgM, VCA IgG, EBNA IgG and heterophile antibodies," which indicates the method used to establish the "ground truth" for those specific samples relative to established serological definitions.

Summary

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NOV 2 5 1998

16982352

510k Summary of Safety and Effectiveness

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.

The assigned 510(k) number is:

Applicant Information:

Date Prepared:	June 11, 1998
Name:	Columbia Bioscience, Inc.
Address:	8775 M Centre Park Drive, #559Columbia, MD 21045

Contact Person:	Norman Jenkins
PhoneNumber:	410-995-0450
Fax Number:	410-995-0448

Device Information:

Trade Name:	EBV-VCA IgM ELISA Kit
Common Name.	EBV-VCA IgM EIA Test
Classification Name;	Epstein-Barr Virus

Equivalent Device: EBV Serology

Device Description: The & EBV-VCA IgM ELISA Kit is an enzyme-linked immunosorbent assay (ELISA) for the detection of IgM antibodies to Epstein-Barr Viral Capsid antigen in human serum.

Intended Use: For the qualitative determination of IgM antibodies in human serum to Epstein Barr (recombinant) Viral Capsid antigen (EBV-VCA) antigen. The EBV-VCA IgM assay should be used in conjunction with other Epstein-Barr serologies (EBV-VCA IgG, EBNA-1 IgG, EA-D IgG, EA-D IgM, EBNA-1 IgM and heterophile) as an aid in the diagnosis of infectious monomucleosis. The test can be performed either manually or in conjunction with the MAGO PLUS ™ Automated EIA Processor.

Principle of Procedure:

The & EBV-VCA IgM ELISA Kit is an enzyme-linked immunosorbent assay to detect IgM to EBV-VCA in human serum. Recombinant EBV-VCA antigen is attached to a solid phase (microtiter well). Diluted test sera are added to each well. If antibodies which recognize the EBV-VCA antigen are present in the patient sample they will bind to the antigen in the well. After incubation, the wells are washed to remove unbound antibody. An enzyme labeled anti-human immunoglobulin (conjugate) is added to each test well. If antibody is present the enzyme-linked antibody will bind to it. After incubation, the wells are washed to remove unbound conjugate. A substrate solution is then added to each well. If enzyme is present from prior step, the reaction is stopped and the color intensity is measured photometrically producing an indirect detection of the specific antibody present in the patient sample.

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Performance Characteristics

A. Clinical Sensitivity and Specificity Using Characterized Sera

Frozen retrospective sera from one hundred and seventy-six patients were characterized using commercially available kits for VCA IgM, VCA IgG, EBNA IgG and heterophile antibodies. Based on the results of this testing, the patient sera were characterized as follows:

- 102 sera were characterized as convalescent (past infection). These were positive for VCA IgG and/or EBNA IgG antibodies and negative for VCA IgM and heterophile antibody.
- 32 scra were characterized as scronegative. These were negative for VCA IgG, VCA IgM, EBNA IgG and beterophile antibody.
- 42 sera were characterized as having a current (recent) infection. These were positive for VCA IgM and/or heterophile antibody and were negative for EBNA IgG.

All 176 sera were then tested by an independent clinical commercial laboratory using the Is-EBV-VCA IgM Test Kit. The results obtained are shown in Table 2:

EBV Serological Status
	Convalescent	Current Infection	Seronegative
POSITIVE	3	38	0
Is-EBV-VCA IgM	NEGATIVE	98	2	32
*EQUIVOCAL	1	2	0
Relative Specificity (Convalescent) 98/101 = 97.0%			95% CI91.6-99.4
Relative Sensitivity (Current Infection) 38/40 = 95.0%			83.1-99.4
Relative Specificity (Seronegative) 32/32 = 100%			89.1-100
Overall Agreement 168/173 = 97.1%			93.4-99.1

Equivocal results were excluded from calculations

NOTE : Please be advised that 'relative' refers to the comparison of the 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's accuracy to predict disease. Since the above studies were performed on a pre-selected, retrospective, population, no calculations for the assay's positive predictive value may be done or inforred.

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B. Precision

To determine the precision of the Is-EBV-VCA IgM Test Kit, four positive and two negative sera were assayed ten times each in three different runs at three different sites. The 3 sites include: the manufacturer, a research and develooment laboratory, and a clinical commercial laboratory. The intra- and interassay precision obtained at each site is shown in Tables 3, 4 and 5. The Inter-Site precision is shown in Table 6.

TABLE 3 : Site #1 - Intra-Assay and Interassay Precision

SERUM	MEANINDEX	INTRA-ASSAY RUN 1CV%	MEANINDEX	INTRA-ASSAY RUN 2CV%	MEANINDEX	INTRA-ASSAY RUN 3CV%	MEANINDEX	INTERASSAYCV%
A (POS)	1.41	8.08	1.77	6.07	1.67	8.45	1.62	12.05
B (POS)	2.17	7.97	2.78	7.01	2.35	6.26	2.44	12.67
C (POS)	4.01	9.92	5.05	7.15	4.22	6.78	4.43	12.86
D (POS)	2.70	8.91	3.32	13.74	2.81	4.35	2.94	13.69
E (NEG)	0.35	17.21	0.54	8.51	0.45	21.03	0.44	22.75
F (NEG)	0.09	49.37	0.17	26.31	0.18	14.93	0.14	39.31
							CAL	1.03	10.81
							PC	1.59	25.45
							NC	0.49	18.70

TABLE 4 : Site #2- Intra-Assay and Interassay Precision

SERUM	INTRA-ASSAY RUN 1		INTRA-ASSAY RUN 2		INTRA-ASSAY RUN 3		INTERASSAY
	MEANINDEX	CV%	MEANINDEX	CV%	MEANINDEX	CV%	MEANINDEX	CV%
A (POS)	1.47	3.96	1.34	4.14	1.31	4.13	1.37	6.53
B (POS)	2.25	6.35	1.99	4.31	2.06	4.90	2.10	7.34
C (POS)	3.92	3.95	3.45	2.43	3.50	4.35	3.62	6.96
D (POS)	2.57	6.26	2.26	5.36	2.41	3.06	2.41	7.31
E (NEG)	0.35	7.41	0.32	8.33	0.33	11.03	0.33	9.38
F (NEG)	0.14	17.42	0.12	10.23	0.12	9.31	0.12	14.80
						CAL	1.00	3.79
						PC	1.43	3.91
						NC	0.19	22.90

			TABLE 5 : Site #3 - Intra-assay and Interassay Precision
--	--	--	----------------------------------------------------------

SERUM	INTRA-ASSAY RUN 1		INTRA-ASSAY RUN 2		INTRA-ASSAY RUN 3		INTERASSAY
	MEAN INDEX	CV%	MEAN INDEX	CV%	MEAN INDEX	CV%	MEAN INDEX	CV%
A (POS)	1.48	4.94	1.43	4.97	1.48	5.15	1.46	5.08
B (POS)	2.19	6.22	2.20	6.44	2.13	4.55	2.17	5.80
C (POS)	3.78	5.99	3.58	4.80	3.70	5.25	3.68	5.69
D (POS)	2.64	5.67	2.52	5.75	2.56	4.15	2.57	5.40
E (NEG)	0.45	12.82	0.38	9.77	0.40	10.27	0.41	13.26
F (NEG)	0.20	9.43	0.16	7.16	0.18	11.82	0.18	12.70
						CAL	1.00	3.22	n=9
						PC	1.21	3.74	n=3
						NC	0.62	1.85	n = 3

						TABLE 6 : Inter-Site Precision
--	--	--	--	--	--	--------------------------------

SERUM(n=90)	INTER-SITE
	MEANINDEX	CV%
A (POS)	1.48	11.13
B (POS)	2.24	11.41
C (POS)	3.91	13.43
D (POS)	2.64	13.09
E (NEG)	0.40	20.69
F (NEG)	0.15	28.77
CAL (n=36)	1.01	6.22
LPC (n=18)	1.42	13.16
NC (n=18)	0.31	60.11

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C. Specificity with Potentially Cross-Reactive Sera

Thirteen scra, reactive for IgM antibodies to varicella zoster, cylomegalovirus and herpes simplex virus by EIA were cested in the Is-EBV-VCA IgM Test Kit. 3/4 anti-VZV IgM positive sera were non-reactive for anti-VCA IgM; 3/5 anti-CMV IgM positive scra were non-reactive for anti-VCA IgM and 4/4 anti-HSV positive scra were non-reactive for anti-VCA IgM. This suggests that some specific cross-reactivity should be expected with the Is-VCA IgM Test Kit from these analytes.

D. Correlation of Manual and MAGO Plus Results

The Is-EBV-VCA IgM Test Kit has been developed for automated as well as manual use. To demonstrate the equivalence of the manual and MAGO Plus procedures, the results of 120 serum samples tested by both methods were plotted. A scattergram and regression line of the results obtained with 95% confidence intervals is shown in Figure 3. The data indicate good correlation with a Pearson Correlation Coefficient of 0.990.

Image /page/3/Figure/4 description: The image is a scatter plot comparing MAGO PLUS INDEX VALUES to MANUAL INDEX VALUES. The x-axis represents MANUAL INDEX VALUES, ranging from 0 to 7, while the y-axis represents MAGO PLUS INDEX VALUES, ranging from -0.5 to 5.5. The plot includes a regression line with the equation Y = 0.0045 + 0.7074X, and the correlation coefficient r is 0.990, indicating a strong positive correlation between the two variables.

FIGURE 3 : Manual and MAGO Plus Result Correlation

D. MAGO Plus Precision

The precision of the assay when performed on the MAGO Plus Automated EIA Processor was determined by assaying six sera ten times each in three different runs. Table 7 shows the intra-and interassay precision obtained using the MAGO Plus.

SERUM	INTRA-ASSAY RUN 1		INTRA-ASSAY RUN 2		INTRA-ASSAY RUN 3		INTERASSAY
	MEAN	CV%	MEAN	CV%	MEAN	CV%	MEAN	CV%
	INDEX		INDEX		INDEX		INDEX
A (POS)	1.15	4.58	1.28	7.18	1.16	9.27	1.20	8.64
B (POS)	1.74	5.55	1.80	6.42	1.69	7.08	1.74	6.68
C (POS)	3.19	4.30	3.34	4.72	3.22	3.53	3.25	4.55
D (POS)	1.96	4.93	2.07	6.85	2.06	5.22	2.03	6.09
E (NEG)	0.28	22.59	0.31	10.20	0.29	19.57	0.29	17.76
F (NEG)	0.10	0.00	0.10	0.00	0.10	0.00	0.10	0.00
						CAL	1.00	4.54	n=S
						PC	1.26	3.20	n = 3
						NC	0.39	9.08	n = 3

TABLE 7 : Site #2- Intra-Assay and Interassay Precision - MAGO Plus

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Image /page/4/Picture/1 description: The image shows the logo for the Department of Health & Human Services USA. The logo consists of a circular border with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES USA" written around it. Inside the circle is an abstract symbol that resembles an eagle or a bird in flight, composed of three curved lines.

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

NOV 2 5 1998

Norman Jenkins President Columbia Bioscience, Inc. 8775 M Centre Park Drive, #559 Columbia, Maryland 21045

Re: K982352 Device: VCA IgM ELISA Test System Dated: September 22, 1998 Received: September 28, 1998

Dear Mr. Jenkins:

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 QS 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.

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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 Sutman

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

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Page 1 of 1

510(k) Number: Not Known

Device Name: 7 VCA IgM ELISA

Indications For Use: For the qualitative determination of IgM antibodies in human serum to Epstein Barr (recombinant) Viral Capsid antigen (VCA) antigen. The 78 VCA IgM assay should be used in conjunction with other Epstein-Barr serologies (VCA IgG, EBNA-1 IgG, EA-D IgM, EA-D IgG, EBNA-1 IgM and heterophile) as an aid in the diagnosis of infectious mononucleosis. The test can be performed either manually or in conjunction with the MAGO PLUS TM Automated EIA Processor

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

Concurrence of CDRH, Office of Device Evaluation (ODE)

Prescription Use X (Per 21 CFR 801.109) OR

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

Woody Dubois

Division of Clinical Laboratory Devices 510(k) Number_1 982352

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).