Quantification of HIV-1 Using Multiple Competitors in a Single-Tube Assay. (August 1996)

• Record Type: Journal Article
• Authors: Vener, T.; Axelsson, M.; Albert, J.; Uhlén, M.; Lundeberg, J.
• Is Part Of: Biotechniques. Volume 21:Number 2(1996)
• Page Start: 248
• Page End: 255
• Publication Date: 1996-08
• Subjects: Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/96212st01
• Languages: English
• ISSNs: 0736-6205

Abstract

Methods for quantification of human immunodeficiency virus type 1 (HIV-1) based on competitive PCR and fragment analysis have been developed. Samples containing HIV-1 DNA and known amounts of three cloned competitors were co-amplified by PCR with semi-nested primers. The competitor DNAs contained the same long terminal repeat primer binding sequences as the wild-type DNA, but they are different in internal sequences and length. One of the inner primers was fluorescent-labeled to allow discrimination between the wild-type DNA and the three competitors by fragment analysis using a standard automated sequencer. A calibration curve using the peak area of the three competitors enabled accurate determination of target amount with minimal variations. The method presented here can be used for quantification of HIV-1 in clinical samples and will be useful for monitoring disease progression and treatment effects.