Quantitation of HIV-1 RNA in plasma using a signal amplification branched DNA (bDNA) assay.

Important note: Information in this article was accurate in 1993. The state of the art may have changed since the publication date.

Quantitation of HIV-1 RNA in plasma using a signal amplification branched DNA (bDNA) assay.

Abstr Gen Meet Am Soc Microbiol. 1993;93:433 (abstract no. T-10). Unique Identifier : AIDSLINE ASM93/93291810
Pachl C; Elbeik T; Saxer M; Kern D; Stempien M; Fong SJ; Sheridan P; Yeghiazarian T; Neuwald P; Urdea M; et al; Chiron Corp., Emeryville, CA.

Abstract: The level of HIV-1 RNA in plasma has been quantitated using a nucleic acid hybridization assay based on signal amplification branched DNA (bDNA) technology. The RNA target is captured onto a microwell surface, detected via bDNA amplifier molecules and alkaline phosphatase labeled probes, and signal is generated using a chemiluminescent substrate. HIV-1 plasma RNA was detected in 81% of seropositive plasma specimens (n = 124), and the frequency of detection at various CD4 cell levels was as follows: 50%, CD4 > 500 (n = 6); 72% CD4 200-500 (n = 58); 93% CD4 < 200 (n = 60). Plasma RNA levels were also analyzed from 12 patients enrolled in the ACTG 175 protocol. Specimens were taken at baseline and 8 weeks after therapy with ddI/AZT, ddC/AZT, AZT or ddI. Consistent values for plasma HIV-1 RNA were observed with two baseline specimens from each patient and viral RNA load stayed the same or decreased in specimens taken 8 weeks after initiation of therapy. Plasma from 6 patients from another study (non-ACTG) were analyzed by quantitative plasma culture and the RNA assay. HIV-1 RNA was detected in all baseline samples whereas only one baseline sample was detected by plasma culture. An increase in viral RNA copies after 4 to 8 weeks correlated with an increase in plasma viremia measured by plasma culture. These results indicate that the HIV-1 RNA assay is a reproducible method for the quantitation of the level of HIV-1 in plasma during the course of antiviral therapy. This method is also amenable to routine laboratory use.

Keywords: Acquired Immunodeficiency Syndrome/*BLOOD/DRUG THERAPY/ MICROBIOLOGY Antigens, CD4/IMMUNOLOGY Antiviral Agents/*THERAPEUTIC USE Biological Markers/BLOOD Didanosine/THERAPEUTIC USE Drug Therapy, Combination Gene Amplification/METHODS Human HIV-1/GENETICS/*ISOLATION & PURIF RNA, Viral/ANALYSIS/*BLOOD T-Lymphocyte Subsets/MICROBIOLOGY Zalcitabine/THERAPEUTIC USE Zidovudine/THERAPEUTIC USE CLINICAL TRIAL ABSTRACT
930930
M9391161

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