Quantitative Detection of Human Immunodeficiency Virus Type 1 (HIV-1) RNA by PCR and Use as a Prognostic Marker and for Evaluating Antiretroviral Therapy

Abstract

The amount of human immunodeficiency virus type 1 (HIV-1) genomic RNA in sera is considered to be one of the most direct markers for patient prognosis and monitoring the efficacy of antiretroviral therapy. Quantitative detection of HIV-1 RNA was performed by the dilution endpoint method and competitive PCR. Conditions for detecting one copy of HIV-1 control DNA were examined to decide the dilution endpoint exactly. PCR of the gag region by SK145/SK39 primer pair and seminested PCR by SK145/SK101 primer pair gave the best result. Conditions for competitive PCR of HIV-1 cDNA, which was reverse transcripted from HIV-1 control RNA, were also studied using a SK38/SK39 primer pair in the presence of HIV-PCR MIMIC as a competitor DNA. The detection limit of HIV-1 control DNA by competitive PCR was 10 copies but that of HIV-1 control RNA was 50 copies. Time-course quantitation of HIV-1 RNA in frozen-stored sera from an AIDS patient was carried out and traced back to 1993. The amount of serum HIV-1 RNA markedly decreased when the treatment was changed but increased again before deterioration of his clinical status. It is considered that the quantitation of serum HIV-1 RNA is useful for the prognosis of HIV-infection and the evaluation of the antiretroviral therapy.