Quantitation of defective and ecotropic viruses during LP-BM5 infection by real time PCR and RT-PCR.
Murine AIDS (MAIDS) is a pathology induced by the LP-BM5 murine leukaemia virus mixture in susceptible strains of mice such as C57BL/6J resulting in lymphoproliferation and progressive immunodeficiency. The etiologic agent of this pathology is BM5d, a replication defective virus. BM5e is a replication competent virus in the viral mixture that functions as a helper virus. This paper describes real time PCR and RT-PCR assays for quantitation of the proviral DNA and viral RNA of BM5d and BM5e. Data is presented describing the change in BM5d and BM5e proviral DNA levels and viral RNA levels in both blood and spleen in the first 8 weeks of infection. Infected mice have increasing levels of BM5d and BM5e viral DNA and RNA detectable from as early as 2 weeks post infection. Similar levels of proviral DNA was found for BM5d and BM5e in PBMC and spleen, however higher levels of BM5e viral RNA were observed in both tissues throughout infection. The assays described can be used as both a diagnostic tool and to investigate the direct effect of treatments on the BM5d and BM5e viruses and MAIDS development.
Paun
Andrea
A
Discipline of Microbiology, School of Biological and Chemical Sciences, The University of Western Australia, Nedlands, WA 6009, Australia.
Shaw
Kathryn
K
Fisher
Scott
S
Sammels
Leanne M
LM
Watson
Mark W
MW
Beilharz
Manfred W
MW
D000818:Animals; D004279:DNA, Viral / Q000032:analysis / Q000097:blood; D003673:Defective Viruses / Q000302:isolation & purification*; D005260:Female; D009052:Leukemia Virus, Murine / Q000302:isolation & purification*; D051379:Mice; D008810:Mice, Inbred C57BL; D016183:Murine Acquired Immunodeficiency Syndrome / Q000209:etiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis / Q000097:blood; D012192:Retroviridae Infections / Q000821:virology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014412:Tumor Virus Infections / Q000821:virology*; D016662:Virus Integration
D004279:DNA, Viral; D012367:RNA, Viral