Biotin-labeled DNA probes for detection of Epstein-Barr virus by in-situ cytohybridization.

Conventional laboratory diagnosis of EBV-related disease is now performed by one of three methods: serology, lymphocyte transformation assay, or Epstein-Barr nuclear antigen (EBNA) staining of cell preparations. Of these techniques, serology is the most widely used. However, this approach assumes an intact host immune system, which is absent or impaired in some of the more baffling EBV-related syndromes. Detection of infectious virus by the lymphocyte transformation assay is labor-intensive, requires access to human umbilical cord blood lymphocytes, and requires a two-month period of incubation. Although detection of EBNA in tissue imprints is rapid, the anticomplement immunofluorescence assay, when applied to clinical materials, is subject to misinterpretation and requires multiple controls. Because of these difficulties, hybridization analysis of clinical materials for presence of EBV with biotinylated DNA probes promises to have wide-ranging applicability in the clinical microbiology laboratory. These techniques can readily be used in other viral systems and have proved useful for detection of human CMV and HSV DNA and RNA. Extension of the techniques to detection of specific nonviral nucleic acid sequences is the next frontier, limited essentially only by definition of significant target sequences.