| Literature DB >> 19917318 |
Carole L Wallis1, Maria A Papathanasopoulos, Shabir Lakhi, Etienne Karita, Anatoli Kamali, Pontiano Kaleebu, Eduard Sanders, Omu Anzala, Linda-Gail Bekker, Gwynn Stevens, Tobias F Rinke de Wit, Wendy Stevens.
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 "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. 2009 Elsevier B.V. All rights reserved.Entities:
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Year: 2009 PMID： 19917318 PMCID： PMC2932961 DOI： 10.1016/j.jviromet.2009.11.011
Source DB: PubMed Journal: J Virol Methods ISSN： 0166-0934 Impact factor: 2.014