Cost-effective pyrosequencing strategies for HIV-1 drug resistance detection

As the duration and size of HIV antiretroviral treatment programmes increase in resource-limited settings, the emergence of transmitted and acquired HIV drug resistance is concerning. This study examined the power of pyrosequencing (PS) on the 454 GS Junior platform to monitor HIV drug resistance pa...

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Bibliographic Details
Published in:Antiviral therapy 2013-01, Vol.18, p.A145-A145
Main Authors: Wallis, C L, St John, E P, Viana, R, Reese, B, Shanbhag, P, Glass, A, Turenchalk, G, Simen, B B
Format: Article
Language:English
Subjects:
Human immunodeficiency virus 1
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Summary:As the duration and size of HIV antiretroviral treatment programmes increase in resource-limited settings, the emergence of transmitted and acquired HIV drug resistance is concerning. This study examined the power of pyrosequencing (PS) on the 454 GS Junior platform to monitor HIV drug resistance patterns via large-scale individual testing or surveillance studies. We demonstrate that 1) a PS-based prototype assay is superior to SS and 2) a PS sample pooling strategy can decrease cost while maintaining accurate surveillance. 80 HIV-1 plasma samples were subjected to Sanger sequencing (SS) and individual PS using prototype primer plates, containing barcoded primers covering PR codons 10-99 and RT codons 1-251. There was a strong correlation between the three protocols. Even a PS mutation detection level of [greater than or equal]20% showed a distinct advantage over SS for haplotype detection. The pool-PS protocol combined with sample barcoding is an affordable high-throughput option for resistant variant detection. Furthermore, PS on samples pooled pre-amplification is a cost-effective option for regional/institutional surveillance.
ISSN:1359-6535