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A rapid zidovudine (ZDV) resistance genotypic assay was developed based on the mutagenically separated PCR (MS-PCR) technique to detect two ZDV-resistant mutations, M41L and K70R in CRF01_AE (subtype E). Endpoint dilution analysis revealed that the newly constructed MS-PCR assay could successfully detect three to nine copies of human immunodeficiency virus type 1 template RNA. The test against wild-type and mutant template mixtures in different ratios demonstrated that the assay could detect 10% minor population, at least. Fifty-one subtype E clinical samples were analyzed by the newly constructed MS-PCR assay and direct nucleotide sequencing. The concordance of the two assays was 92 and 100% in codons 41 and 70, respectively. The MS-PCR assay is a rapid, simple, and inexpensive assay that is highly sensitive in detecting mutant targets, including minor populations. Thus, it could be used as a powerful tool for epidemiological surveillance of drug-resistant mutations in developing countries.

Original publication

DOI

10.1128/aac.46.12.3861-3868.2002

Type

Journal article

Journal

Antimicrob Agents Chemother

Publication Date

12/2002

Volume

46

Pages

3861 - 3868

Keywords

Base Sequence, DNA Mutational Analysis, Drug Resistance, Viral, Genotype, HIV-1, Humans, Polymerase Chain Reaction, Zidovudine