Methods for sensitive detection of reverse transcriptase

A method for differentiating between infection by human immunodeficiency virus-1 Group M and Group O in a human immunodeficiency virus-1 infected subject comprising the steps of:

• a) contacting a biological sample from the subject with a ribonucleotide of SEQ ID NO:4, an oligonucleotide of SEQ ID NO:2 and a Group M reverse transcriptase inhibiting amount of nevirapine under conditions whereby the oligonucleotide and the ribonucleotide will anneal and a DNA strand will be synthesized as an extension from the oligonucleotide if reverse transcriptase in the sample is not inhibited by the nevirapine;
• b) amplifying the synthesized DNA by the polymerase chain reaction whereby the conditions of the amplification comprise about 35 cycles of heating the synthesized DNA and a primer pair consisting of the oligonucleotide of SEQ ID NO:10 and the oligonucleotide of SEQ ID NO:2 to about 95.degree. C. for one minute, about 55.degree. C. for one minute and about 72.degree. C. for one minute;
• and detecting the amplification of the synthesized DNA, the amplification of the synthesized DNA indicating the presence of human immunodeficiency virus-1 Group O in the sample.