The immunoperoxidase technique for rapid human cytomegalovirus identification

Abstract

Sixteen cytomegalovirus (CMV) isolates from both ill and healthy patients were identified by the immunoperoxidase technique (IP). CMV detection was accomplished by direct examination of the primary isolate using either direct (DIP) or indirect immunoperoxidase (IIP) techniques. In thirteen of the isolates, confirmation of identification was achieved by indirect immunofluorescence (IFA) and by demonstration of herpes particles by electron microscopy (EM). Further, in four cases of non-CMV alterations in the tissue culture which might be confused with actual infection, the IP test was negative as were the confirmatory tests. The IIP is preferred over the DIP test since the latter shows a certain amount of background stain of uninfected cells. Tissue culture cells showing focal CMV cytopathic effect contained both nuclear and cytoplasmic inclusions stained by IP technique. Nonspecific staining was associated with cytoplasmic inclusion bodies. The IP technique can detect individual cell CMV infection at 24 hours when EM reveals only unenveloped viral particles. It is sensitive, specific, and allows direct identification of infected cells in the primary isolate in as little as 90 minutes. Furthermore, it can be performed in standard isolation tissue culture tubes, whereas IFA requires the transfer of the infected cells onto slides or the routine use of Leighton tubes.