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BioMed Research International
Volume 2017 (2017), Article ID 4979252, 6 pages
Research Article

A Robust PCR Protocol for HIV Drug Resistance Testing on Low-Level Viremia Samples

1Department of Medical Microbiology, University of Manitoba, Winnipeg, MB, Canada
2The Bioinformatics Core, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, MB, Canada
3National HIV & Retrovirology Laboratories, JC Wilt Infectious Diseases Research Center, Public Health Agency of Canada, Winnipeg, MB, Canada
4Department of Biochemistry and Medical Genetics, University of Manitoba, Winnipeg, MB, Canada
5Cadham Provincial Laboratory, Winnipeg, MB, Canada
6Department of Pediatrics and Child Health, University of Manitoba, Winnipeg, MB, Canada

Correspondence should be addressed to Hezhao Ji

Received 21 January 2017; Revised 14 March 2017; Accepted 21 March 2017; Published 3 April 2017

Academic Editor: György Schneider

Copyright © 2017 Shivani Gupta et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


The prevalence of drug resistance (DR) mutations in people with HIV-1 infection, particularly those with low-level viremia (LLV), supports the need to improve the sensitivity of amplification methods for HIV DR genotyping in order to optimize antiretroviral regimen and facilitate HIV-1 DR surveillance and relevant research. Here we report on a fully validated PCR-based protocol that achieves consistent amplification of the protease (PR) and reverse transcriptase (RT) regions of HIV-1 pol gene across many HIV-1 subtypes from LLV plasma samples. HIV-spiked plasma samples from the External Quality Assurance Program Oversight Laboratory (EQAPOL), covering various HIV-1 subtypes, as well as clinical specimens were used to optimize and validate the protocol. Our results demonstrate that this protocol has a broad HIV-1 subtype coverage and viral load span with high sensitivity and reproducibility. Moreover, the protocol is robust even when plasma sample volumes are limited, the HIV viral load is unknown, and/or the HIV subtype is undetermined. Thus, the protocol is applicable for the initial amplification of the HIV-1 PR and RT genes required for subsequent genotypic DR assays.