Detection of feline immunodeficiency virus transcripts by quantitative reverse transcription-polymerase chain...

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.