[Development of an ultrasensitive mismatch tolerant PCR assay for the qualitative and quantitative determination of HIV-1 RNA].

A real-time fluorescent polymerase chain reaction was used to develop a RealBest HIV RNA kit that was clinically suitable for the detection of HIV-1 RNA and for the estimation of virus load in plasma and serum samples. Due to the selection of a highly conserved target region and to the experimental study of the impact of different primer-template and probe-template mismatches on RT-PCR with subsequent selection of the optimum oligonucleotide set, the developed assay can detect and measure the concentration of all subtypes of HIV-1, group M. The assay provides a high reproducibility and sensitivity and a wide dynamic range of virus loads (20 to 10 million IU/ml of plasma or serum).