A robust PCR protocol for HIV drug resistance testing on low-level viremia samples
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Date
2017-04-03
Authors
Gupta, Shivani
Taylor, Tracy
Patterson, Aileen
Liang, Binhua
Bullard, Jared
Sandstrom, Paul
Domselaar, Gary Van
Ji, Hezhao
Journal Title
Journal ISSN
Volume Title
Publisher
BioMed Research International
Abstract
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 theHIV 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.
Description
Keywords
HIV, low-level viremia, PCR, Drug resistance