Detection of feline immunodeficiency virus transcripts by quantitative reverse transcription-polymerase chain reaction.
We developed a quantitative reverse transcription-polymerase chain reaction (RT-PCR) procedure to estimate replication status of feline immunodeficiency virus (FIV) in peripheral blood mononuclear cells (PBMCs) of cats. Primers used for the RT-PCR were designed to detect only multiple spliced transcripts of FIV and allowed us to detect the specific transcripts with high specificity. By using limiting-cell-dilution RT-PCR, we demonstrated that the specific transcripts were quantitatively detected in a single infected cell in a background of 1 x 10(6) uninfected cells without Southern blot hybridization. Furthermore, the transcripts were observed efficiently in all PBMCs of the chronically FIV-infected cats when examined by this RT-PCR technique. There results demonstrated that this RT-PCR method is applicable for specific detection of the FIV-specific transcripts in the PBMCs and for estimation of the viral replication status in vivo.
Tomonaga
K
K
Department of Veterinary Microbiology, University of Tokyo, Japan.
Mikami
T
T
D000818:Animals; D001483:Base Sequence; D002415:Cats; D002460:Cell Line; D017931:DNA Primers; D005814:Genes, Viral; D016089:Immunodeficiency Virus, Feline / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000821:virology*; D008214:Lymphocytes / Q000821:virology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014158:Transcription, Genetic
D017931:DNA Primers