Optimization of the oligonucleotide ligation assay, a rapid and inexpensive test for detection of HIV-1 drug resistance mutations, for non-North American variants

Optimization of the oligonucleotide ligation assay, a rapid and inexpensive test for detection of HIV-1 drug resistance mutations, for non-North American variants Beck I. A. auteur aut IRD Crowell C. auteur aut IRD Kittoe R. auteur aut IRD Bredell H. auteur aut IRD Machaba M. auteur aut IRD Willamson C. auteur aut IRD Janssens W. auteur aut IRD Jallow S. auteur aut IRD van der Groen G. auteur aut IRD Shao Y. auteur aut IRD Jacob M. auteur aut IRD Samuel N. M. auteur aut IRD de Rivera I. L. auteur aut IRD Ngo-Giang-Huong Nicole auteur aut IRD Cassol S. auteur aut IRD Alemnji G. auteur aut IRD Frenkel L. M. auteur aut IRD text journalArticle eng text/pdf born digital access Objective: We evaluated the feasibility of the oligonucleotide ligation assay (OLA), a specific, sensitive, and economical ligase-based point mutation assay designed to detect HIV-1 drug-resistance mutations at 12 codons of HIV-1 subtype B pol, for potential use in resource-poor settings. Methods: Specimens from HIV-1-infected individuals collected by 7 international laboratories, including subtypes A, B, C, D, F, G, J, and recombinants AE and AG, were tested by the OLA developed for HIV-1 subtype B. Common polymorphisms that interfered with reactivity of the OLA were identified and modified probes designed and evaluated. Results: 92.5% (2410) of 2604 codons in specimens from 217 individuals were successfully genotyped by the subtype B OLA. A high rate (range 8.3%-31.2%) of indeterminate results (negative OLA reaction for both mutant and wild type) was observed for 5 codons. Modified probes at reverse transcriptase codons 151 and 184 and protease codon 90 increased the rate of valid OLA to 96.1%. Conclusions: The OLA designed for HIV-1 subtype B genotyped most pol codons in non-B subtypes from Asia and Africa but was improved by addition of several modified probes. International laboratories experienced in molecular techniques were able to perform the OLA. specialized Hiv drug resistance Hiv-1 non-b subtypes Resistance testing Oligonucleotide ligation assay Minor genotypes Point mutation assay Dried blood spots 052 050 JAIDS Journal of Acquired Immune Deficiency Syndromes 48 4 418-427 2008 1525-4135 https://www.documentation.ird.fr/hor/fdi:010042699 10.1097/QAI.0b013e31817ed7d7 1525-4135 [F B010042699] https://www.documentation.ird.fr/hor/fdi:010042699 https://www.documentation.ird.fr/intranet/publi/2008/08/010042699.pdf Accès réservé (Intranet de l'IRD) IRD - Base Horizon / Pleins textes 2008-09-19 2017-08-23 fdi:010042699 fre