An affordable HIV-1 drug resistance monitoring method for resource limited settings.

Authors:
Dr. Justen Manasa, HBMLS, MPhil, PhD
Dr. Justen Manasa, HBMLS, MPhil, PhD
University of Zimbabwe
Senior Lecturer
Molecular Biology, Virology, Bioinformatics
Harare, Harare | Zimbabwe
Siva Danaviah
Siva Danaviah
University of KwaZulu-Natal
Sureshnee Pillay
Sureshnee Pillay
University of KwaZulu-Natal
Prevashinee Padayachee
Prevashinee Padayachee
University of KwaZulu-Natal
South Africa
Hloniphile Mthiyane
Hloniphile Mthiyane
University of KwaZulu-Natal
Richard John Lessells
Richard John Lessells
University of KwaZulu-Natal
Berea | South Africa
Christopher Seebregts
Christopher Seebregts
Biomedical Informatics Research

J Vis Exp 2014 Mar 30(85). Epub 2014 Mar 30.

Africa Centre for Health and Population Studies, College of Health Sciences, University of KwaZulu-Natal, Durban, South Africa;

HIV-1 drug resistance has the potential to seriously compromise the effectiveness and impact of antiretroviral therapy (ART). As ART programs in sub-Saharan Africa continue to expand, individuals on ART should be closely monitored for the emergence of drug resistance. Surveillance of transmitted drug resistance to track transmission of viral strains already resistant to ART is also critical. Unfortunately, drug resistance testing is still not readily accessible in resource limited settings, because genotyping is expensive and requires sophisticated laboratory and data management infrastructure. An open access genotypic drug resistance monitoring method to manage individuals and assess transmitted drug resistance is described. The method uses free open source software for the interpretation of drug resistance patterns and the generation of individual patient reports. The genotyping protocol has an amplification rate of greater than 95% for plasma samples with a viral load >1,000 HIV-1 RNA copies/ml. The sensitivity decreases significantly for viral loads <1,000 HIV-1 RNA copies/ml. The method described here was validated against a method of HIV-1 drug resistance testing approved by the United States Food and Drug Administration (FDA), the Viroseq genotyping method. Limitations of the method described here include the fact that it is not automated and that it also failed to amplify the circulating recombinant form CRF02_AG from a validation panel of samples, although it amplified subtypes A and B from the same panel.

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Source
http://dx.doi.org/10.3791/51242DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4024245PMC

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March 2014
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