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

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• 作者：Carole L. Wallis ; Maria A. Papathanasopoulos ; Shabir Lakhi ; Etienne Karita ; Anatoli Kamali ; Pontiano Kaleebu ; Eduard S ; ers ; Omu Anzala ; Linda-Gail Bekker ; Gwynn Stevens ; Tobias F. Rinke de Wit ; Wendy
• 关键词：HIV-1 subtype C ; Antiretroviral drug resistance ; Mutation profile ; Affordable
• 刊名：Journal of Virological Methods
• 出版年：2010
• 出版时间：February 2010
• 年：2010
• 卷：163
• 期：2
• 页码：505-508
• 全文大小：554 K

文摘

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.