Direct enzyme-linked immunosorbent assay that uses peroxidase-labeled antigen for determination of immunoglobulin M antibody to cytomegalovirus

Abstract

A direct enzyme-linked immunosorbent assay was developed for the measurement of immunoglobulin M (IgM) antibody to cytomegalovirus (CMV). Wells of microtiter plates were coated with anti-human IgM. Each patient's serum was added at a dilution of 1:100, and IgM from the serum was allowed to react with anti-human IgM. The amount of CMV-specific IgM antibody bound was determined by measuring the intensity of color change after the addition of peroxidaselabeled CMV antigen and substrate. Nuclei of infected cells served as an antigen source. CMV IgM could be detected only in IgM fractions of sera from patients with a recent CMV infection. Rheumatoid factor did not cause false-positive results. No cross-reactions were observed when paired sera from 22 patients with herpes simplex or varicella and single sera from 12 patients with suspected infectious mononucleosis were tested by the direct enzyme-linked immunosorbent assay. Each of 17 patients with a seroconversion for CMV antibody showed CMVspecific IgM antibody. In six of these patients the antibody was detected in the initial serum. The direct enzyme-linked immunosorbent assay for CMV IgM is a specific and sensitive test for the diagnosis of recent CMV infections and possesses distinct advantages over indirect tests. 18 to 72 years of age. Four patients were children 3, 4, 9, and 15 years of age, none of whom had been congenitally infected with CMV. In addition, sera were obtained from 34 patients clinically suspected of having herpes simplex or varicella-zoster infection or infectious mononucleosis. Furthermore, 36 sera with 416

An enzyme labelled nuclear antigen immunoassay for detection of cytomegalovirus IgM antibodies in human serum: Specific and non‐specific reactions

“…Our results concerning the diagnostic value of CMV-specific IgM are in good agreement with those obtained by others [Cappel et al, 1978;van Loon et al, 1981;Kangro et al 1982;Pass et al, 1983;Sutherland and Briggs, 19831. IgM was detectable in all cases of true primary and in 73% of secondary infections.…”

Detection of cytomegalovirus‐specific igm in renal transplant recipients

“…In our study, the ELA-M assay performed better than the IFA-M test in both sensitivity and specificity, confirming earlier studies using an IgM capture technique (30)(31)(32). Interference from rheumatoid factors, antinuclear antibodies, and immune complexes have been encountered in other studies utilizing immunofluorescence assays (33); similar problems with specificity of the IFA-M were found in our study. The ELA-M assay gave consistent results without the pretreatment of the sample to remove IgG.…”

A novel Western blot test containing both viral and recombinant proteins for anticytomegalovirus immunoglobulin M detection

“…Serum IgM to HCMV can be detected by a variety of different procedures, but its detection has been hampered by several technical problems causing interassay variability. The most widely used procedure for IgM detection is enzyme immunoassay (EIA) (18,(24)(25)(26)(27). Many different EIAs for HCMV IgM are available commercially, and poor agreement has been found among the results obtained with different EIA kits (13).…”