POLYMERASE CHAIN-REACTION DETECTION OF TYPE-D SIMIAN RETROVIRUS PROVIRAL DNA FROM INFECTED MACAQUES

A simple polymerase chain reaction (PCR) approach was developed for de tection of Type D simian retrovirus (SRV) serogroup 2 proviral DNA usi ng peripheral blood lymphocytes (PBLs) obtained from infected macaques . PCR primer pairs were developed against serogroup 2 envelope (enu) g ene sequence, and fidelity of PCR fragment amplification was determine d using molecularly cloned SRV serogroup 2 (D2/RHE/OR) DNA, and genomi c DNA from Raji cells independently infected with different SRV serogr oups. One primer pair exhibiting high fidelity was then utilized for P CR detection of serogroup 2 proviral DNA from PBLs, and from cells sor ted into immune cell subpopulations by fluorescent-activated cell sort ing (FAGS). Enu PCR fragments were readily detected from as few as 10( 4) PBLs or immune cell subpopulations. In addition, highly specific PC R primers against serogroups 1 and 3 were utilized to detect proviral DNA from Raji cells infected with SRV serogroups. In all cases, primer s designed to amplify serogroups 1, 2, and 3 proviral DNA were specifi c for their intended serogroup. This primer information and developmen t of a PCR approach for detection of specific SRV proviral DNA will. b e of potential utility as a rapid surveillance tool in monitoring type D simian retrovirus infection within Asian macaque colonies.