A SENSITIVE METHOD FOR THE DETECTION OF MURINE C-TYPE RETROVIRUSES

A RT-PCR assay was developed for group-specific detection of murine C- type retroviruses using a nested set of degenerated primers. To distin guish exogenous viruses from related, but silent endogenous viruses, a DNAse I pretreatment of supernatants is applied. This is followed by a heat inactivation/denaturation step. The PCR method is ultrasensitiv e, which enables the detection of 100 attogram of MoMuLV proviral DNA or up to 1-10 infectious mouse C-type retroviruses in 10 mu l supernat ant of infected cells. The high specificity of the method allows the d ifferentiation between mouse C-type retroviruses and related retroviru ses of the A, B, and D type and C-type retroviruses found in other spe cies. It serves as a valuable tool for the screening of animal cell cu ltures for contaminations with mouse retroviruses, e.g. hybridomas or recombinant cell lines producing foreign proteins. Copyright (C) 1997 Elsevier Science B.V.