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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
medicine
A pragmatic approach to HIV-1 drug resistance determination in resource-limited settings by use of a novel genotyping assay targeting the reverse transcriptase-encoding region only
Journal of Clinical Microbiology, Volume 51, No. 6, Year 2013
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Description
In resource-limited settings (RLS), reverse transcriptase (RT) inhibitors form the backbone of first-line treatment regimens. We have developed a simplified HIV-1 drug resistance genotyping assay targeting the region of RT harboring all major RT inhibitor resistance mutation positions, thus providing all relevant susceptibility data for first-line failures, coupled with minimal cost and labor. The assay comprises a one-step RT-PCR amplification reaction, followed by sequencing using one forward and one reverse primer, generating double-stranded coverage of RT amino acids (aa) 41 to 238. The assay was optimized for all major HIV-1 groupMsubtypes in plasma and dried blood spot (DBS) samples using a panel of reference viruses for HIV-1 subtypes A to D, F to H, and circulating recombinant form 01-AE (CRF01-AE) and applied to 212 clinical plasma samples and 25 DBS samples from HIV-1-infected individuals from Africa and Europe. The assay was subsequently transferred to Uganda and applied locally on clinical plasma samples. All major HIV-1 subtypes could be detected with an analytical sensitivity of 5.00E+3 RNA copies/ml for plasma and DBS. Application of the assay on 212 clinical samples from African subjects comprising subtypes A to D, F to H (rare), CRF01-AE, and CRF02-AG at a viral load (VL) range of 6.71E+2 to 1.00E+7 (median, 1.48E+5) RNA copies/ml was 94.8% (n = 201) successful. Application on clinical samples in Uganda demonstrated a comparable success rate. Genotyping of clinical DBS samples, all subtype C with a VL range of 1.02E+3 to 4.49E+5 (median, 1.42E+4) RNA copies/ml, was 84.0% successful. The described assay greatly reduces hands-on time and the costs required for genotyping and is ideal for use in RLS, as demonstrated in a reference laboratory in Uganda and its successful application on DBS samples. Copyright © 2013, American Society for Microbiology. All Rights Reserved.
Authors & Co-Authors
Aitken, Susan C.
Netherlands, Utrecht
University Medical Center Utrecht
Bronze, Michelle
South Africa, Johannesburg
University of the Witwatersrand
Wallis, Carole Lorraine
South Africa, Johannesburg
Lancet Laboratories
Stuyver, Lieven J.
Belgium, Beerse
Janssen Pharmaceutica, Headquarters
Steegen, Kim
South Africa, Johannesburg
University of the Witwatersrand
South Africa, Johannesburg
National Health Laboratory Service
Balinda, Sheila N.
Uganda, Kampala
Joint Clinical Research Center Uganda
Kityo, Cissy Mutuluuza
Uganda, Kampala
Joint Clinical Research Center Uganda
Stevens, Wendy Susan
South Africa, Johannesburg
University of the Witwatersrand
South Africa, Johannesburg
National Health Laboratory Service
Rinke de Wit, Tobias Floris
Netherlands, Amsterdam
Amsterdam Institute for Global Health and Development
Netherlands, Amsterdam
Pharmaccess International
Schuurman, Rob J.
Netherlands, Utrecht
University Medical Center Utrecht
Statistics
Citations: 26
Authors: 10
Affiliations: 8
Identifiers
Doi:
10.1128/JCM.00118-13
ISSN:
00951137
e-ISSN:
1098660X
Research Areas
Cancer
Health System And Policy
Infectious Diseases
Study Locations
Uganda