Detection of RNA in purified cytomegalovirus virions

Abstract

Five viral RNA transcripts have recently been detected in purified virions of human cytomegalovirus (HCMV) strain AD169, a well-characterized member of the family Herpesviridae [Science 288 (2000) 2373]. While the function of these transcripts and/or the proteins they encode remains to be elucidated, it is not known whether these transcripts are unique to strain AD169 or are present in other HCMV strains. The objective of this study was to determine if these RNAs are present in other HCMV laboratory strains (Towne and Davis), and a low passage clinical isolate (CL203). These strains of CMV were purified by sequential ultracentrifugation through 20% d-sorbitol and glycerol–potassium tartarate gradients and the morphology and infectivity of the virions confirmed by electron microscopy and inoculation into cell culture. When RNA extracted from the purified virions was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) the UL 21.5 and TRL/IRL 2–5 transcripts were detected in virions of HCMV strains AD169, Davis, Towne and CL203. The presence of the UL 21.5 and TRL/IRL 2–5 RNA transcripts in all strains tested demonstrates that the packaged transcripts occurs in all strains of HCMV suggesting that they may have a relevant role in the biology of this virus.