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open-access-imgOpen AccessQuantification of HIV-1 Using Multiple Competitors in a Single-Tube Assay
Author(s)
Tanya Vener,
Mikael D. Axelsson,
Jan Albert,
Mathias Uhlén,
Joakim Lundeberg
Publication year1996
Publication title
biotechniques
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
Language(s)English
SCImago Journal Rank0.617
H-Index131
ISSN0736-6205
DOI10.2144/96212st01

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