Varnier OE, Giri AA, McDermott JL, Martini I, Giacomini M, Campelli A, Puppo F, Indiveri F, Klotman ME, Cara A; Conference on Retroviruses and Opportunistic Infections.
Program Abstr 6th Conf Retrovir Oppor Infect Conf Retrovir Oppor Infect 6th 1999 Chic Ill. 1999 Jan 31-Feb 4; 6th: 101 (abstract no. 177).
Molecular Virology Unit, ABCr, University of Genova, Italy.
Our strategy for the detection of the various forms of HIV DNA includes a quantitative amplification of Total DNA and qualitative amplification of Non-Integrated 2LTR DNA from the same cell lysate (4x10(5)PBMCs). Both biotinylated amplicons are hybridized, captured into streptavidin coated microplates, and detected colorimetrically using the Quanti-Kin(c) software. Total DNA was detected in 315 out of 318: 92 had 50-100 copies, 106 samples 100-1,000 and 26 >1,000 copies. Less than 50 copies were found in 60 samples, while DNA was observed in the remaining 31 samples only using a sensitive protocol. Sequential samples showed 2 distinct profiles of Total DNA: 1) <500 copies with a linear trend, 2) >500 copies with wide fluctuations. The analysis of 101 samples, obtained from untreated or treated patients with non-detectable (responder) or quantifiable (non-responder) HIV RNA, suggests that Total DNA is a marker for therapy efficacy. The circular 2LTR Non-Integrated DNA was detected in samples collected before the start of therapy or in a viremic period of therapeutical inefficacy, while it was not detectable during HAART. These results suggest that Total DNA is a marker for therapy efficacy. The 2LTR Non-Integrated DNA is present during active viral replication and ex novo infection of naive lymphocytes; thus its undetectability indirectly suggests that Total DNA is made by Integrated DNA.
Publication Types:
Keywords:
- Antiretroviral Therapy, Highly Active
- Branched DNA Signal Amplification Assay
- DNA
- DNA, Viral
- HIV Infections
- Humans
Other ID:
UI: 102188820
From Meeting Abstracts