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.
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 x 10(5) 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 approximately 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.
Vandegraaff
N
N
National Centre for HIV Virology Research, Infectious Diseases Laboratories, Institute of Medical and Veterinary Science, Adelaide, Australia.
Kumar
R
R
Burrell
C J
CJ
Li
P
P
D002460:Cell Line; D004279:DNA, Viral / Q000032:analysis*; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D007700:Kinetics; D016133:Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000821:virology*; D016662:Virus Integration; D014779:Virus Replication
D004279:DNA, Viral