Journal of Virological Methods, 28 (1990) 293-298 Elsevier 293 VIRMET 01022 Rapid antiviral DNA-DNA hybridization assay for human cytomegalovirus Wayne M. Dankner’, David Scholl 3,4, Sylvia C. Stanat’, Michael MartinI, Robert L. Sonke’ and Stephen A. Spector’>* ‘Department of Pediatrics and ‘Center for Molecular Genetics, University of California, San Diego, U.S.A., 3Department of Zoology and Biomedical Sciences, Ohio University, U.S.A., 4Diagnostic Hybrids, Inc. and ‘Department of Virology, Burroughs W ellcome Co., U.S.A. (Accepted 22 February 1990) A rapid DNA-DNA hybridization technique that can be accomplished in 4 to 5 days was compared with plaque reduction assay to determine its reliability in per- forming antiviral assays for human cytomegalovirus (HCMV). The assay involves lysing infected cells, direct wicking of denatured DNA onto membranes and hybridization using a 1251-labeled HCMV DNA probe. Using ten ganciclovir sensi- tive clinical HCMV strains for comparison, the DNA hybridization technique correlated well with the plaque assay. Clinical HCMV strains previously identified as resistant to ganciclovir were also readily identified. The DNA-DNA hybridi- zation assay is less tedious and more rapid than plaque reduction assays, and thus, provides an excellent alternative for evaluation of the antiviral activity of drugs against HCMV. Human cytomegalovirus; DNA-DNA hybridization; Ganciclovir; Antiviral assay Introduction Human cytomegalovirus (HCMV) is an important pathogen in neonates, trans- plant patients and in individuals with acquired immunodeficiency syndrome (Nank- ervis and Kumar, 1978; Betts, 1982; Jacobson and Mills, 1988). The severe diseases associated with HCMV infection emphasize the need to develop effective antiviral Correspondence to: Stephen A. Spector, UCSD Medical Center, 225 Dickinson Street, H-814-H. San Diego, CA 92103, U.S.A. 016f50934/90/$03.50 @ 1990 Elsevier Science Publishers B.V. (Biomedical Division)