Abstract
Restriction enzyme digestion analysis of cytomegalovirus (CMV) isolates is used as a powerful tool to study the epidemiology of CMV infections. Use of this technique is hampered by the necessity of growing sufficient virus in tissue culture which may take 3-6 months. Using 32 EcoRI clones from the AD169 strain of CMV, our studies of the CMV genome show that polymorphism is present in the fragments closest to the long and the short inverted repeat sequences of the joint regions. Complete heterogeneity is present for all different CMV strains when hybridized to joint pieces, F and H. Using these observations, we developed a rapid method to determine molecular relatedness of different clinical isolates of CMV. DNA is extracted directly from primary cultures or clinical specimens containing CMV, cleaved with restriction enzyme EcoRI, separated by electrophoresis, transferred to nitrocellulose filters, hybridized to 32P-labeled fragment F or H, and exposed to x-ray film. Using this procedure, we find that all random isolates of CMV exhibit different fragments hybridizing to clones F and H, while isolates showing identical restriction enzyme digestion patterns exhibit identical DNA bands hybridizing to the joint fragments. This procedure reduces the need to passage CMV in tissue culture, can be performed in less than a week, and should greatly simplify studies designed to define the epidemiology of CMV infections.
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Spector, S., Hirats, K. & Neuman, T. RAPID DETERMINATION OF MOLECULAR RELATEDNESS OF CLINICAL CYTOMEGALOVIRUS STRAINS. Pediatr Res 18 (Suppl 4), 286 (1984). https://doi.org/10.1203/00006450-198404001-01160
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DOI: https://doi.org/10.1203/00006450-198404001-01160