Quantitation of HIV-1 DNA with a sensitive TaqMan assay that has broad subtype specificity.
The increasing diversity of HIV-1 isolates makes virus quantitation challenging, especially when diverse isolates co-circulate in a geographical area. Measuring the HIV-1 DNA levels in cells has become a valuable practical tool for fundamental and clinical research. A quantitative HIV-1 DNA assay was developed based on TaqMan(®) technology. Primers that target the highly conserved LTR region were designed to detect a broad array of HIV-1 variants, including viral isolates from many subtypes, with high sensitivity. Introduction of a pre-amplification step prior to the TaqMan(®) reaction allowed the specific amplification of fully reverse transcribed viral DNA. Execution of the pre-amplification step with a second primer set enables for the exclusive quantitation of the 2-LTR circular HIV-1 DNA form.
Laboratory of Experimental Virology, Department of Medical Microbiology, Centre for Infection and Immunity Amsterdam, Academic Medical Centre, University of Amsterdam, Meibergdreef 15, Amsterdam, The Netherlands.
Renée M
RM
van der Sluis
Thijs
T
van Montfort
Mireille
M
Centlivre
Nick C T
NC
Schopman
Marion
M
Cornelissen
Rogier W
RW
Sanders
Ben
B
Berkhout
Rienk E
RE
Jeeninga
William A
WA
Paxton
Georgios
G
Pollakis
D002460:Cell Line; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics; D014644:Genetic Variation; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D016133:Polymerase Chain Reaction; D012367:RNA, Viral; D012680:Sensitivity and Specificity; D013601:T-Lymphocytes / Q000821:virology; D019914:Taq Polymerase
D017931:DNA Primers; D004279:DNA, Viral; D012367:RNA, Viral; D019914:Taq Polymerase