Affordable in-house antiretroviral drug resistance assay with good performance in non-subtype B HIV-1.

Journal article

The introduction of antiretroviral (ARV) therapy in resource-poor settings is effective in suppressing HIV-1 replication and prolonging life of infected individuals. This has led to a demand for affordable HIV-1 drug resistance assays, since treatment failure due to development of drug resistance is common. This study developed and evaluated an affordable "in-house" genotyping assay to monitor HIV-1 drug resistance in Africa, particularly South Africa. An "in-house" assay using automated RNA extraction, and subtype C specific PCR and sequencing primers was developed and successfully evaluated 396 patient samples (viral load ranges 1000-1.6 million RNA copies/ml). The "in-house" assay was validated by comparing sequence data and drug resistance profiles from 90 patient and 10 external quality control samples to data from the ViroSeq HIV-1 Genotyping kit. The "in-house" assay was more efficient, amplifying all 100 samples, compared to 91 samples using Viroseq. The "in house" sequences were 99.2% homologous to the ViroSeq sequences, and identical drug resistance mutation profiles were observed in 96 samples. Furthermore, the "in-house" assay genotyped 260 of 295 samples from seven African sites, where 47% were non-subtype C. Overall, the newly validated "in-house" drug resistance assay is suited for use in Africa as it overcomes the obstacle of subtype diversity.

Authors: Wallis, C; Papathanasopoulos, M; Lakhi, S; Karita, E; Kamali, A

• Publication status: Published
• Journal: Journal of virological methods
• Volume: 163
• Issue: 2
• Pages: 505-508
• Publication date: 2010-02-01
• DOI: 10.1016/j.jviromet.2009.11.011
• EISSN: 1879-0984
• ISSN: 0166-0934
• Language: English
• Keywords: RNA, Viral; Sensitivity and Specificity; Automation; South Africa; Genes, Viral; Genotype; Microbial Sensitivity Tests; Reverse Transcriptase Polymerase Chain Reaction; Drug Resistance, Viral; Humans; HIV-1; Anti-Retroviral Agents; DNA Primers