title: A sensitive method for the detection of murine C-type retroviruses.
abstract: A RT-PCR assay was developed for group-specific detection of murine C-type retroviruses using a nested set of degenerated primers. To distinguish 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 ultrasensitive. which enables the detection of 100 attogram of MoMuLV proviral DNA or up to 1-10 infectious mouse C-type retroviruses in 10 microl supernatant of infected cells. The high specificity of the method allows the differentiation between mouse C-type retroviruses and related retroviruses of the A, B, and D type and C-type retroviruses found in other species. It serves as a valuable tool for the screening of animal cell cultures for contaminations with mouse retroviruses, e.g. hybridomas or recombinant cell lines producing foreign proteins.
authors.affiliation: Department of Regulation and Differentiation, Gesellschaft für Biotechnologische Forschung, Braunschweig, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Heinemeyer
authors.forename: A
authors.initials: A
authors.lastname: Klingenhoff
authors.forename: W
authors.initials: W
authors.lastname: Hansen
authors.forename: L
authors.initials: L
authors.lastname: Schumacher
authors.forename: H
authors.initials: H
authors.lastname: Hauser
authors.forename: M
authors.initials: M
authors.lastname: Wirth
mesh_terms: D016475:3T3 Cells; D000818:Animals; D002460:Cell Line; D004279:DNA, Viral / Q000032:analysis; D017988:Gammaretrovirus / Q000235:genetics / Q000302:isolation & purification*; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D004279:DNA, Viral