Evaluation and Design of Affordable and Novel HIV-1 Drug Resistance Assays:

[摘要] Approximately 5 million individuals are infected with HIV/AIDS in South Africa. TheSouth African government has initiated a National Anti-retroviral therapy (ARV) Programto manage this disease. The emergence of drug resistance to ARV therapy is of greatconcern. Commercial gold standard sequence-based genotyping assays for monitoringresistance are unaffordable.This project aimed at developing affordable methods to detect specific point mutationsrelevant to HIV-1 subtype C. The Oligonucleotide Ligation assay (OLA), a real-time PCRassay and a Restriction Fragment Length Polymorphism (RFLP) assay were explored.Results were compared to the Viroseq genotyping assay. OLA performed poorly on HIV-1subtype C samples and needs modification. The real-time PCR assay using short MinorGroove Binding probes, accurately detected the K65R mutation. The Mae III RFLP assaydetected all V106M mutations accurately. Longitudinal cohort studies are required toconfirm relevant mutations, appropriate assays and algorithms for resistance monitoring inHIV-1 subtype C.

[发布机构] University of the Witwatersrand