Title: Affordable in-house antiretroviral drug resistance assay with good performance in non-subtype B HIV-1
Authors: Wallis CL, Papathanasopoulos MA, Lakhi S, Karita E, Kamali A, Kaleebu P, Sanders E, Anzala O, Bekker LG, Stevens G, De Wit TF, Stevens W .
Journal: J Virol Methods,163:505-508 (2010)
Abstract
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 âinâhouseâ genotyping assay to monitor HIV-1 drug resistance in Africa, particularly South Africa. An âin-houseâ assay using automated RNA extraction, and subtype C specific PCR and sequencing primers was developed and successfully evaluated 396 patient samples (viral load ranges 1000â1.6 million RNA copies/ml). The âin-houseâ 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 âin-houseâ assay was more efficient, amplifying all 100 samples, compared to 91 samples using Viroseq. The âin houseâ sequences were 99.2% homologous to the ViroSeq sequences, and identical drug resistance mutation profiles were observed in 96 samples. Furthermore, the âin-houseâ assay genotyped 260 of 295 samples from seven African sites, where 47% were non-subtype C. Overall, the newly validated âin-houseâ drug resistance assay is suited for use in Africa as it overcomes the obstacle of subtype diversity.
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Citation: Wallis CL, Papathanasopoulos MA, Lakhi S, Karita E, Kamali A, Kaleebu P, Sanders E, Anzala O, Bekker LG, Stevens G, De Wit TF, Stevens W . Affordable in-house antiretroviral drug resistance assay with good performance in non-subtype B HIV-1 J Virol Methods,163:505-508 (2010).