A more precise HIV integration assay designed to detect small differences finds lower levels of integrated DNA in HAART treated patients.
Many studies report the level of total viral DNA in HIV-infected patients, but few studies report the level of integrated DNA. It is important to measure integrated DNA in HIV-infected patients because the information could shed light on the effectiveness of antiretroviral therapy, especially intensified therapy, when viral loads may remain undetectable. In order to develop an integration assay for patient samples, we enhanced the sensitivity of our prior integration assay. To do this, we exploited a technique that we developed, called repetitive sampling, and optimized reaction conditions for rare event detection, rather than large dynamic range. We also designed our primers to match more conserved regions of HIV. The result is a new, sensitive, quantitative assay that allows us to measure integrated DNA in HIV-infected patients. When we applied our integration assay to patient PBMCs, we found that the use of HAART is associated with reduced levels of integrated DNA.
Yu
Jianqing J
JJ
Department of Pathology and Laboratory Medicine, Division of Transfusion Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
Wu
Te Lang
TL
Liszewski
Megan K
MK
Dai
Jihong
J
Swiggard
William J
WJ
Baytop
Clifford
C
Frank
Ian
I
Levine
Bruce L
BL
Yang
Wei
W
Theodosopoulos
Theodore
T
O'Doherty
Una
U
D019380:Anti-HIV Agents / Q000627:therapeutic use*; D023241:Antiretroviral Therapy, Highly Active; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D006678:HIV / Q000187:drug effects*; D015658:HIV Infections / Q000188:drug therapy* / Q000821:virology*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000821:virology; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D016662:Virus Integration / Q000187:drug effects*
D019380:Anti-HIV Agents; D017931:DNA Primers; D004279:DNA, Viral