1415Z
1415Z
1415Z
EBV-EA IgG
Cat # 1415Z
* Laboratory results can never be the only base of a medical report. The patient history and further tests have to be
taken into account
DAI Code # 11 1
NAME AND INTENDED USE
The DIAGNOSTIC AUTOMATION Epstein Barr Virus Early Antigen (EBV-EA) IgG Enzyme-linked
Immunosorbent Assay (ELISA), is intended for the detection of IgG antibody to Epstein Barr Virus Nuclear
Antigen-1 in human sera and plasma.
immunosuppressed patients, pregnant women, and persons of advanced age. Antibodies to the R component may
be found at moderate to high levels in patients with Burkitt’s lymphoma. In contrast, patients with
nasopharyngeal carcinoma may produce high titer antibodies to the D component. Elevated levels of anti-EA and
IgG anti-VCA may be detected in patients with chronic or recurrent illness suspected of being caused by EBV8.
However, a diagnosis of chronic EBV should not be based on the presence of antibodies to EA since elevated
anti-EA titers may also be found in patients with other diseases as well as in healthy individuals with past EBV
infections 5.
DAI Code # 11 2
MATERIAL PROVIDED
1. Microwell strips: EBV-EA antigen coated wells. (12 x 8 wells)
2. Sample Diluent: Blue Color solution 1 vial (22 ml)
3. Calibrator: Factor value (f) stated on label. Red Cap. 1 vial (150 µl)
4. Negative control: Range stated on label. Natural Cap. 1 vial (150 µl)
5. Positive control: Range stated on label. Green Cap. 1 vial (150 µl)
6. Washing concentrate 10x: White Cap. 1bottle (100 ml)\
7. Enzyme conjugate: Red color solution. 1 vial (12 ml)
8. TMB Chromogenic Substrate: Amber bottle. 1 vial (12 ml)
9. Stop solution: 2 N HCl. 1 vial (12 ml)
DAI Code # 11 3
ASSAY PROCEDURE
1. Place the desired number of coated strips into the holder.
2. Prepare 1:20 dilutions by adding 10 µl of the samples, negative control, positive control, and calibrator to
200 µl of sample diluent. Mix well.
3. Dispense 100 µl of diluted sera, calibrator, and controls into the appropriate wells. For the reagent blank,
dispense 100 µl sample diluent in 1A well position. Tap the holder to remove air bubbles from the liquid and
mix well. Incubate for 30 minutes at room temperature.
4. Remove liquid from all wells. Repeat washing three times with washing buffer.
5. Dispense 100 µl of enzyme conjugate to each well and incubate for 30 minutes at room temperature.
6. Remove enzyme conjugate from all wells. Repeat washing three times with washing buffer.
7. Dispense 100 µl of TMB Chromogenic Substrate to each well and incubate for 30 minutes at room
temperature.
8. Add 100 µl of 2 N HCl to stop reaction.
Make sure there are no air bubbles in each well before reading.
9. Read O.D. at 450 nm with a microwell reader.
CALCULATION OF RESULTS
1. To obtain Cut off OD value: Multiply the OD of Calibrator by Factor (f) printed on
label of Calibrator.
2. Calculate the EBV-EA IgG Index of each determination by dividing the OD values
of each sample by obtained OD value of Cut off.
For example:
If Factor (f) value on label = 0.4
Obtained Calibrator O.D. = 1.100
Cut-off O.D. = 1.100 x 0.4 = 0.44 (By definition EBV-EA IgG Index = 1)
Patient sample O.D. = 0.580
EBV-EA IgG Index = 0.580 / 0.44 = 1.32 (Positive result)
Patient sample O.D. = 0.320
EBV-EA IgG Index = 0.320 / 0.44 = 0.73 (Negative result)
QUALITY CONTROL
The test run may be considered valid provided the following criteria are met:
1. The O.D. value of the reagent blank against air from a microwell reader should be less than 0.150.
2. If the O.D. value of the Calibrator is lower than 0.250, the test is not valid and must be repeated.
3. The EBV-EA IgG Index for Negative and Positive Control should be in the range stated on the labels.
INTERPRETATION
DAI Code # 11 4
PERFORMANCE CHARACTERISTICS
Histogram:
215 random samples are determined with DIAGNOSTIC AUTOMATION Microwell ELISA EBV-EA IgG. The
test results are computed as IgG Index using a chosen reference serum as IgG Index 1. The distribution of
frequency versus IgG Index value is presented as following:
P / N Ratio
63.2% (136 samples) have IgG index below 1.
Mean value = 0.509 SD = 0.229
IgG index 1 (cut off value) = Mean value + 2 SD
Precision:
The precision of the assay was evaluated by testing three different sera of eight replicates over a period of one
week. The intra-assay and inter-assay C.V. are summarized below:
DAI Code # 11 5
LIMITATIONS OF THE ASSAY
1. The values obtained from this assay are intended to be an aid to diagnosis only. Each physician must interpret
the results in light of the patient’s history, physical findings and other diagnostic procedures.
2. Results from children should be reviewed with caution. This kit is designed to measure IgG antibody in
patient samples. Positive results in neonates must be interpreted with caution, since maternal IgG is
transferred passively from the mother to the fetus before birth.
3. Results obtained from immunocompromised individuals should be interpreted with caution.
4. There is a possibility of assay cross-reactivity with specimens containing anti-E.coli antibody.
REFERENCES
1. Epstein, M.A., B.B. Achong, and Y.M. Barr. 1964. Virus particles in Cultured Lymphoblasts from Burkitt’s
Lymphoma. In: Lancet 1:702-703.
2. Epstein, M.A., Y.M. Barr, and B.G. Achong. 1965. Studies with Burkitt’s Lymphoma. In: Wistar Inst.
Sympos. Monogr. 4:69-82.
3. Schooley, R.T. and R. Dolin. 1985. Epstein-Barr Virus Infectious Mononucleosis). In: Principles and
Practice of Infectious Diseases, 2nd Edition. Mandell, G.L., R.G. Douglas, and J.E. Bennett. (eds). John
Wiley and Sons, New York. pp 97-982.
4. Lennette, E.T. 1988. Herpesviridae: Epstein-Barr Virus. In: Laboratory Diagnosis of Infectious Diseases,
Principles and Practice. Vol. Ll. Lennette, E.H., Halonen, P., Murphy, F.A., eds. Springer-Verlag, NY. Pp.
230-246.
5. Lennette, E.T. and W. Henle. 1987. Epstein-Barr Virus Infections: Clinical and Serological Features.
Laboratory Management June: pp. 22-28.
6. Henle G., W. Henle, and C.A. Horwitz. 1974. Antibodies to Epstein-Barr Virus-Associated Nuclear Antigen
in Infection Mononucleosis. Journal of infectious Diseases. 130:231-239.
7. Lennette ET, and Henle W: Epstein-Barr virus infections: Clinical and Serologic features. Lab Mgmt. 23-28,
June, 1987.
8. Joncas J, Lapointe N, Gervais F, Leyritz M, and Wills A: Unusual prevalence of antibodies to Epstein-Barr
virus early antigen in ataxia telangiectasia. Lancet 1:1160, 1977.
9. Akaboshi I, Jamamoto J, Katsuki T, and Matsuda l: Unique pattern of Epstein-Barr virus specific antibodies
in recurrent parotitis. Lancet 2:1049-1051, 1983.
DAI Code # 11 6