Kinetics of Human Immunodeficiency Virus Type 1 (HIV) DNA Integration in Acutely Infected Cells as Determined Using a Novel Assay for Detection of Integrated HIV DNA

doi:10.1128/JVI.75.22.11253-11260.2001

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Journal of Virology, November 2001, p. 11253-11260, Vol. 75, No. 22
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.22.11253-11260.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Kinetics of Human Immunodeficiency Virus Type 1 (HIV) DNA Integration in Acutely Infected Cells as Determined Using a Novel Assay for Detection of Integrated HIV DNA

Nick Vandegraaff,¹^,² Raman Kumar,¹ Christopher J. Burrell,¹^,² and Peng Li¹^,^*

National Centre for HIV Virology Research, Infectious Diseases Laboratories, Institute of Medical and Veterinary Science,¹ and Department of Molecular Biosciences, University of Adelaide,² Adelaide, Australia

Received 11 December 2000/Accepted 17 August 2001

We have developed a novel linker-primer PCR assay for the detection and quantification of integrated human immunodeficiencyvirus type 1 (HIV) DNA. This assay reproducibly allowed the detectionof 10 copies of integrated HIV DNA, in a background of 2 × 10⁵ cell equivalents of human chromosomal DNA, without amplifyingextrachromosomal HIV DNA. We have used this assay and a near-synchronousone-step T-cell infection model to investigate the kinetics ofviral DNA accumulation following HIV infection. We report herethat integrated HIV DNA started accumulating 1 h after the firstappearance of extrachromosomal viral DNA and accounted for ~10%of the total HIV DNA synthesized in the first round of viral replication.These results highlight the efficient nature of integrase-mediatedHIV integration in infected Tcells.

^* Corresponding author. Mailing address: National Centre for HIV Virology Research, Infectious Diseases Laboratories, Instituteof Medical and Veterinary Science, Frome Rd., Adelaide 5000, Australia.Phone: 61 8 82223544. Fax: 61 8 82223543. E-mail: peng.li{at}imvs.sa.gov.au.

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