Quantitative determination of cytomegalovirus IgG antibody by enzyme-linked immunosorbent assay (ELISA)

Abstract

Quantitative determination of cytomegalovirus (CMV) human IgG antibody by ELISA performed in polystyrene trays was carried out and compared with results obtained by the routine complement fixation (CF) test. The antibody titres of all the sera determined by ELISA were higher than those detected by the CF test. Of the 27 sera negative (less than 1:8) by the CF test, 22 of them were also negative (less than 1:50) by ELISA. The other five CF negative sera were found to have low levels of antibody by ELISA. ELISA is a sensitive and specific assay for CMV antibody. The CMV antigen adsorbed readily to the microtitre plates and the test was simple to perform. The results of ELISA could be obtained in 5 to 6h. A description of the test procedure is given including the method of CMV antigen attachment to the plates. The possibility of using paired sera at a single dilution in the ELISA test to detect seroconversion is discussed.

Antibody to cytomegalovirus in patients with sjögren's syndrome. As determined by an enzyme‐linked immunosorbent assay

“…The antigen preparations (purchased from Flow Laboratories, Inglewood, CA) were cytomegalovirus strain AD169, cytomegalovirus control antigen consisting of lysed fibroblasts of the type in which the virus had been grown, mumps virus, and mumps virus control antigen consisting of parallel chorioallantoic membrane preparations in which the virus had been grown. This cytomegalovirus antigen has been used successfully in the ELISA in other laboratories (10).…”

Detection of immunoglobulin M and G antibodies against cytomegalovirus early and late antigens by enzyme-linked immunosorbent assay

“…before CF, hence allowing a very early diagnosis of acute CMV infection. DISCUSSION In recent years, a number of papers have described the development of ELISA technology for the detection of antibodies against CMV in human sera (4,7,18,20,26,27,30,35,36,39,40). However, despite the fact that antigen quality is a major determinant in the specificity and reproducibility of the technique (2,15), very little attention has been paid to the production of CMV antigens suitable for use in the ELISA.…”

“…The measurement of IgM, however, is complicated by falsepositive reactions due to the presence of rheumatoid factor (RF), thus requiring its removal before testing for specific IgM (34). Recently, a number of new serological techniques have been described for the detection of antibodies against CMV, either based upon the principles of a radioimmunoassay (11,24,36) or an enzyme-linked immunosorbent assay (ELISA) (4,7,18,20,26,27,30,35,36,39), which appear to be both rapid and sensitive. This would justify their application for routine serology (2,3,13).…”

“…However, the detection of anti-CMV-EA antibodies by these techniques has not been described as yet, and comparisons of radioimmunoassay or ELISA data with those of a sensitive serological test such as IF are scarce (13,18,40). In most instances, comparisons have been made with relatively insensitive, although still widely used, tests such as complement fixation (CF) or indirect hemagglutination inhibition (3,5,7,26,28). A major advantage of ELISAs and radioimmunoassays compared with IF is the possibility to quantify the results by using automated equipment for reading the results and a table top computer for data processing (8,17,25).…”

“…Such an approach for routine serology requires standardization of all procedures and ingredients, including the antigens used (2). This aspect has been relatively neglected in previous reports on ELISA for CMV (4,7,18,30,36,39,40).…”