Quantitation of HIV-1 DNA with a sensitive TaqMan assay that has broad subtype specificity

Renée M. van der Sluis; Thijs van Montfort; Mireille Centlivre; Nick C. T. Schopman; Marion Cornelissen; Rogier W. Sanders; Ben Berkhout; Rienk E. Jeeninga; William A. Paxton; Georgios Pollakis

doi:https://doi.org/10.1016/j.jviromet.2012.09.019

Quantitation of HIV-1 DNA with a sensitive TaqMan assay that has broad subtype specificity

Renée M. van der Sluis, Thijs van Montfort, Mireille Centlivre, Nick C. T. Schopman, Marion Cornelissen, Rogier W. Sanders, Ben Berkhout, Rienk E. Jeeninga, William A. Paxton, Georgios Pollakis

Research output: Contribution to journal › Article › Academic › peer-review

15 Citations (Scopus)

Abstract

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

Original language	English
Pages (from-to)	94-102
Journal	Journal of virological methods
Volume	187
Issue number	1
DOIs	https://doi.org/10.1016/j.jviromet.2012.09.019
Publication status	Published - 2013

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https://doi.org/10.1016/j.jviromet.2012.09.019

Cite this

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title = "Quantitation of HIV-1 DNA with a sensitive TaqMan assay that has broad subtype specificity",

abstract = "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({\textregistered}) 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({\textregistered}) 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",

author = "{van der Sluis}, {Ren{\'e}e M.} and {van Montfort}, Thijs and Mireille Centlivre and Schopman, {Nick C. T.} and Marion Cornelissen and Sanders, {Rogier W.} and Ben Berkhout and Jeeninga, {Rienk E.} and Paxton, {William A.} and Georgios Pollakis",

year = "2013",

doi = "https://doi.org/10.1016/j.jviromet.2012.09.019",

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journal = "Journal of virological methods",

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T1 - Quantitation of HIV-1 DNA with a sensitive TaqMan assay that has broad subtype specificity

AU - van der Sluis, Renée M.

AU - van Montfort, Thijs

AU - Centlivre, Mireille

AU - Schopman, Nick C. T.

AU - Cornelissen, Marion

AU - Sanders, Rogier W.

AU - Berkhout, Ben

AU - Jeeninga, Rienk E.

AU - Paxton, William A.

AU - Pollakis, Georgios

PY - 2013

Y1 - 2013

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AB - 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

U2 - https://doi.org/10.1016/j.jviromet.2012.09.019

DO - https://doi.org/10.1016/j.jviromet.2012.09.019

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SN - 0166-0934

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