A Simple Method to Distinguish Between Simian Immunodeficiency Virus Isolates by Restriction Analysis of PCR Products

Abstract

A simple method to distinguish between simian immunodeficiency virus (SIV) isolates of experimentally infected rhesus macaques is reported. Peripheral blood mononuclear cells (PBMC) were prepared from a rhesus macaque infected with SIV_StM isolated originally from a stump-tailed macaque, or from a rhesus monkey infected with SIV_SM from a sooty mangabey monkey. PBMC were cocultivated with CEM × 174 cells and a region of the SIV envelope (env) gene was amplified by the polymerase chain reaction (PCR) from cDNA of infected cocultivation cells. Restriction enzyme digestion analysis of the PCR products enabled SIV_StM and SIV_SM to be differentiated from each other, and from a molecular clone of SIV_MAC, SIV_MAC239, originally isolated from an infected rhesus macaque. Furthermore, when SIV_SM and SIV_StM were introduced into the same animal, restriction enzyme analysis of the PCR product amplified from cocultivation cells of this rhesus macaque suggested that the animal was superinfected.