Detection of diverse variants of human immunodeficiency virus-1 groups M, N, and O and simian immunodeficiency viruses from chimpanzees by using generic pol and env primer pairs

Abstract

Human immunodeficiency virus type 1 (HIV-1) infection of humans is the result of independent cross-species transmissions of simian immunodeficiency viruses (SIVcpz) from naturally infected chimpanzees (Pan troglodytes troglodytes) to man. To develop a polymerase chain reaction-based assay capable of detecting members of all major phylogenetic SIVcpz and HIV-1 lineages (groups M, N, and O), primer pairs in conserved pol and env regions were designed. Both primer sets amplified </=10 copies of selected group M reference clones (subtypes A-H), proviral DNA or RNA of group N (YBF30), and group O of HIV-1 and also amplified divergent SIVcpz from cultured isolates (SIVcpzGAB1 and SIVcpzANT), uncultured spleen tissue (SIVcpzUS), and plasma (SIVcpzANT and SIVcpzUS). Sequences of the 2 amplicons (445 bp for gp41 and 261 bp for integrase) are of sufficient length for phylogenetic analyses, allowing both group and subtype classifications of the human viruses. Finally, both primer pairs are highly sensitive (>99%) in amplifying viral sequences from plasma taken from patients infected with HIV-1 group M (n=226) and O (n=17) viruses.