Immunoperoxidase technique for detection of antibodies to human cytomegalovirus

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Abstract
The indirect immunoperoxidase antibody technique (IPA) was applied to determine immunoglobulin (Ig)G to human cytomegalovirus (CMV) antibodies in 114 blood donor sera, 4 cases of congenital cytomegalic inclusion disease, and 4 cases of acquired CMV infection. The results were compared with those obtained with CMV complement fixation (CF) and indirect fluorescent antibody technique (IFA) for broad spectrum CMV antibody (.SIGMA.Ab) detection. IgG antibody was detected by CF and IPA. In healthy adults IPA and IFA .SIGMA.Ab titers are usually higher than CF titers. Some sera negative by CF and IPA are positive at low dilutions by IFA .SIGMA.Ab antibody determination, due to the detection of small amounts of IgA or noncomplement-fixing IgG. Nonspecific results seem unlikely, since only nuclear inclusion fluorescence was interpreted as specific, as demonstrated by blocking tests. In acute CMV infection, the IFA .SIGMA.Ab and IPA IgG titers are essentially the same, except during the 1st weeks of infection, when IFA titers are higher and IgM is detectable. No cross-reactivity with other herpes group viruses, herpes simplex and varicella-zoster, was observed. Although some problems of nonspecific staining of cytoplasmic inclusions are shared by both methods, the IPA technique seems to possess the same degree of sensitivity and specificity as the IFA technique, but interpretation is easier and various procedural steps can be delayed without the technical problems associated with fluorescence microscopy.