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 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.
Heinemeyer
T
T
Department of Regulation and Differentiation, Gesellschaft für Biotechnologische Forschung, Braunschweig, Germany.
Klingenhoff
A
A
Hansen
W
W
Schumacher
L
L
Hauser
H
H
Wirth
M
M
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
D004279:DNA, Viral