DETECTION OF FELINE IMMUNODEFICIENCY VIRUS PROVIRAL DNA IN FELINE PERIPHERAL-BLOOD MONONUCLEAR-CELLS BY THE NESTED 2-STEP POLYMERASE CHAIN-REACTION

The polymerase chain reaction (PCR) was applied to detect feline immun odeficiency virus (FIV) proviral DNAs in primary periphal blood mononu clear cells (PBMC). Suitable conditions for PCR amplification were exa mined to obtain highly sensitive and specific results by simple staini ng in agarose gel. Specific amplification of FIV proviral DNA in PBMC DNA of FIV-infected cats was achieved by a nested two-step PCR that am plified the DNA first with outer primers and then with inner primers n ested within the first primers. PCR amplification using different prim ers indicated that those based on the gag sequence of the FlV/TM2 stra in isolated in Japan were suitable for the detection of FIV genomes in naturally infected Japanese pet cats. By the nested two-step PCR with mixed gag primers of TM2 and Petaluma, isolated in the USA, we could detect FIV genomes in all 11 primary PBMC samples from FIV-seropositiv e cats tested. The PCR protocol developed here is sensitive and specif ic for molecular detection of FIV infection in cats.