A Single-tube real-time RT-PCR assay for RNA detection and quantification of genetically diverse HIV including rare HIV-1 non-m groups and HIV-2

Abstract

The RT-PCR method with real-time fluorescence detection was used for the development of a prototype of a diagnostic kit for reliable diagnosis of genetic variants of RNA of HIV-1 groups M, N, O, and P and RNA of HIV-2 in blood plasma and serum. The kit is tolerant to nucleotide mismatches, provides a broad linear range of concentrations of HIV RNA, 100% analytical specificity, and adequate analytical sensitivity: 42 ME/mL (HIV-1 group M), 45 copies/mL (HIV-2), 92 copies/mL (HIV-1 group O), and 90 copies/mL (HIV-1 group N). The kit provided successful detection and measurement of HIV RNA concentration in all the samples of the international reference panel for HIV-1 genotypes. The results of the test correlate with the results of commercial tests.