Detection of guinea pig cytomegalovirus nucleic acids in cultured cells with biotin-labelled hybridization probes.

Abstract

Biotin labelled hybridization probes prepared from recombinant plasmids containing segments of the guinea pig cytomegalovirus (GPCMV) genome were used to detect GPCMV nucleic acids in guinea pig cells by in situ hybridization. The time course of GPCMV infection was assessed in two cultured cell types, guinea pig embryo (GPE) cells and 104C1 cells, a transformed and cloned guinea pig cell line. Detection of GPCMV nucleic acids was accomplished in both cell types with individual GPCMV DNA fragments and with mixtures of GPCMV DNA fragments. When compared to other established methods of GPCMV detection, the method of in situ hybridization enabled the detection of a higher percentage of positive cells early during the course of the infection. In addition, differences in the replication cycle of GPCMV in the two cultured cell lines could be demonstrated. These findings will facilitate future studies of GPCMV tissue tropism in vivo.