DIRECT QUANTIFICATION OF HIV-1 RNA BY CAPILLARY ELECTROPHORESIS WITH LASER-INDUCED FLUORESCENCE

HIV-1 RNA was quantitated directly by capillary electrophoresis with l aser-induced fluorescence (CE-LIF). CE-LIF was used to analyze cellula r RNA and various nucleotide complexes. A fluorescently labeled DNA pr obe (DNA/RNA complex) in conjunction with thiazole orange intercalator was determined to have optimal stability and sensitivity for RNA anal ysis. Based on this observation, a hybridization method using a HIV-sp ecific fluorescently labeled probe with analysis by CE-LIF was develop ed. Plasma samples from a HIV-seropositive patient were lysed to obtai n RNA, hybridized with the HIV-specific probe and analyzed by CE-LIF. As little as 19 fg (1710 copies per 1 mi of starting plasma) of HIV RN A can be reliably and quantitatively detected. CE-LIF appears to be an efficient and sensitive method to quantitatively analyze RNA from a v ariety of sources. (C) 1997 Elsevier Science B.V.