Gupta P, Ding M, Cottrill M, Rinaldo C, Mellors J; National Conference on Human Retroviruses and Related Infections.

Program Abstr Second Natl Conf Hum Retrovir Relat Infect Natl Conf Hum Retrovir Relat Infect 2nd 1995 Wash DC. 1995 Jan 29-Feb 2; 119.

University of Pittsburgh, Pittsburgh, PA.

Quantitation of HIV-1 DNA and RNA in cells and body fluids has become an important tool in studies of HIV-1 pathogenesis and antiretroviral efficacy. Current quantitative competitive PCR (QC PCR) assays are cumbersome, requiring 10-12 reactions per sample. We have developed a novel internally controlled PCR (ICPCR) to quantitate HIV-1 gag DNA and RNA in PBMC and plasma that requires only two reactions per sample. Samples are spiked with 500 copies of HIV gag DNA or RNA containing a deletion in the target sequence. The PCR products derived from full-length or deleted DNA/RNA can be easily differentiated by gel retardation. The amount of HIV-1 DNA/RNA is quantitated by interpolation from a standard curve generated with known copies of deleted gag DNA/RNA. The ICPCR assay is sensitive and reproducible within the lienar range of amplification of 10-10(3) copies of HIV-1 DNA/RNA. The assay detected HIV-1 RNA in plasma from all HIV-infected subjects regardless of disease stage. The level of HIV-1 RNA was inversely correlated with CD4 cell number. ICPCR ws compared with bDNA in subjects initiating antiretroviral therapy. The reduction in HIV-1 RNA response to therapy was comparable with the two assays.

Publication Types:

• Meeting Abstracts

Keywords:

• Acquired Immunodeficiency Syndrome
• Base Sequence
• Biological Assay
• DNA
• DNA Primers
• Genes, gag
• HIV
• HIV Core Protein p24
• HIV Infections
• HIV Seropositivity
• HIV-1
• Polymerase Chain Reaction
• RNA
• RNA, Viral
• genetics
• immunology
• reverse transcriptase, Human immunodeficiency virus 1

Other ID:

• 95920383

UI: 102213332

From Meeting Abstracts