Simple detection of point mutations associated with HIV-1 drug resistance.

Journal article

A novel assay is described for the detection of HIV-1 drug resistance that is simple, cheap and sensitive. HIV-1 drug resistance in B and non-B HIV-1 subtypes was investigated using Mutagenically-Separated PCR (MS--PCR) --- a competitive semi-nested PCR which uses mutagenic primers. The assay was assessed for sensitivity, specificity and its ability to detect mutant virus within a mixed mutant--wild-type population. Gene sequencing was carried out simultaneously for comparison. MS--PCR detected five copies of HIV-1 RNA from laboratory isolates and 50 copies from patient samples. We demonstrate 100% specificity of detection for wild type or mutant virus for clades A, B, C, D and E. For mixed populations of virus, MS--PCR can detect at least a 10% mix of wild type:mutant, or vice-versa. When applied to African patient samples MS--PCR detected 91.6% of the codons tested. Concordance with sequencing data was 88.8% for protease and 97.2% for RT. MS--PCR is sensitive and specific for the detection of mutations in HIV-1, and can be adapted easily to test for resistance at any codon of interest.

Authors: Frater, A; Chaput, C; Beddows, S; Weber, J; McClure, M

• Publication status: Published
• Journal: Journal of virological methods
• Volume: 93
• Issue: 1-2
• Pages: 145-156
• Publication date: 2001-04-01
• DOI: 10.1016/s0166-0934(01)00266-x
• EISSN: 1879-0984
• ISSN: 0166-0934
• Language: English
• Keywords: Drug Resistance, Multiple; Point Mutation; Anti-HIV Agents; Africa; Sensitivity and Specificity; Drug Resistance, Microbial; Humans; HIV Infections; Polymerase Chain Reaction; Antiretroviral Therapy, Highly Active; HIV-1