An HIV-1 integrase genotype assay for the detection of drug resistance mutations

Sexual Health
Anna C HearpsSuzanne M Crowe

Abstract

The integrase inhibitors (e.g. Raltegravir) are a new class of antiretroviral drugs that have recently become available for the treatment of patients with multi-drug resistant HIV-1 within Australia. The emergence of mutations that confer resistance to the integrase inhibitors has been observed in vivo; however, no commercial genotyping assay is currently available to screen for resistance to these drugs. The HIV-1 integrase gene was amplified from plasma-derived HIV-1 viral RNA via reverse transcription-polymerase chain reaction and genotype determined via population DNA sequencing. Drug resistance mutations and polymorphisms were detected using the Stanford University online HIV database. Assay sensitivity and reproducibility were determined using clinical and laboratory-derived samples. Our in-house assay was capable of genotyping the integrase gene from all samples tested (n = 30) of HIV-1 subtypes B, C, D, F, CFR01_AE and CRF02_AG and can amplify the integrase region from plasma samples containing as few as 50 HIV RNA copies/mL. The assay is highly reproducible (average nucleotide concordance = 99.6%, n = 4) and is capable of detecting resistance-associated mutations. This assay is suitable for routine drug resistance scre...Continue Reading

References

Jan 30, 2008·Antimicrobial Agents and Chemotherapy·Isabelle MaletAnne-Geneviève Marcelin
Jul 25, 2008·The New England Journal of Medicine·Roy T SteigbigelUNKNOWN BENCHMRK Study Teams
Jul 25, 2008·The New England Journal of Medicine·David A CooperUNKNOWN BENCHMRK Study Teams
Aug 20, 2008·Expert Opinion on Pharmacotherapy·Patricia Marr, Sharon Walmsley
Feb 18, 2009·The Journal of Antimicrobial Chemotherapy·Isabelle MaletAnne-Genevieve Marcelin

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