TY - JOUR
T1 - Direct quantification of HIV-1 RNA by capillary electrophoresis with laser-induced fluorescence
AU - Kolesar, J. M.
AU - Allen, P. G.
AU - Doran, C. M.
PY - 1997
Y1 - 1997
N2 - HIV-1 RNA was quantitated directly by capillary electrophoresis with laser-induced fluorescence (CE-LIF) CE-LIF was used to analyze cellular RNA and various nucleotide complexes. A fluorescently labeled DNA probe (DNA/RNA complex) in conjunction with thiazole orange intercalator was determined to have optimal stability and sensitivity for RNA analysis. Based on this observation, a hybridization method using a HIV-specific fluorescently labeled probe with analysis by CE-LIF was developed. Plasma samples from a HIV-seropositive patient were lysed to obtain RNA, hybridized with the HIV-specific probe and analyzed by CE-LIF. As little as 19 fg (1710 copies per 1 ml of starting plasma) of HIV RNA can be reliably and quantitatively detected. CE-LIF appears to be an efficient and sensitive method to quantitatively analyze RNA from a variety of sources.
AB - HIV-1 RNA was quantitated directly by capillary electrophoresis with laser-induced fluorescence (CE-LIF) CE-LIF was used to analyze cellular RNA and various nucleotide complexes. A fluorescently labeled DNA probe (DNA/RNA complex) in conjunction with thiazole orange intercalator was determined to have optimal stability and sensitivity for RNA analysis. Based on this observation, a hybridization method using a HIV-specific fluorescently labeled probe with analysis by CE-LIF was developed. Plasma samples from a HIV-seropositive patient were lysed to obtain RNA, hybridized with the HIV-specific probe and analyzed by CE-LIF. As little as 19 fg (1710 copies per 1 ml of starting plasma) of HIV RNA can be reliably and quantitatively detected. CE-LIF appears to be an efficient and sensitive method to quantitatively analyze RNA from a variety of sources.
KW - Gene quantification
KW - Human immunodeficiency virus (HIV)
KW - RNA
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U2 - 10.1016/S0378-4347(97)00334-4
DO - 10.1016/S0378-4347(97)00334-4
M3 - Article
C2 - 9342669
AN - SCOPUS:0030981698
SN - 1572-6495
VL - 697
SP - 189
EP - 194
JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
IS - 1-2
ER -