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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 immunodeficiency virus type 1 (HIV) DNA. This assay reproducibly allowed the detection of 10 copies
of integrated HIV DNA, in a background of 2 × 105
cell equivalents of human chromosomal DNA, without amplifying extrachromosomal HIV DNA. We have used this assay and a
near-synchronous one-step T-cell infection model to investigate the
kinetics of viral DNA accumulation following HIV infection. We report
here that integrated HIV DNA started accumulating 1 h after the
first appearance 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-mediated HIV integration in infected T cells.
*
Corresponding author. Mailing address: National Centre
for HIV Virology Research, Infectious Diseases Laboratories, Institute of 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.
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.
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