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open-access-imgOpen AccessQuantification of HIV-1 Using Multiple Competitors in a Single-Tube Assay
Tanya Vener,
Mikael D. Axelsson,
Jan Albert,
Mathias Uhlén,
Joakim Lundeberg
Publication year1996
Publication title
Resource typeJournals
PublisherFuture Science Ltd
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.
Subject(s)biology , chemistry , dna , dna sequencer , dna sequencing , gene , genetics , human immunodeficiency virus (hiv) , microbiology and biotechnology , organic chemistry , polymerase chain reaction , primer (cosmetics) , virology
SCImago Journal Rank0.617

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