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


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 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.

SUBMITTER: Gupta S 

PROVIDER: S-EPMC5394351 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

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

Gupta Shivani S   Taylor Tracy T   Patterson Aileen A   Liang Binhua B   Bullard Jared J   Sandstrom Paul P   Van Domselaar Gary G   Ji Hezhao H  

BioMed research international 20170403


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 <i>pol</i>  ...[more]

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