Optimization of the oligonucleotide ligation assay, a rapid and inexpensive test for detection of HIV-1 drug resistance mutations, for non-North American variants.
| Autor: | Beck IA; Division of Infectious Diseases and Virology, University of Washington, Seattle, WA, USA., Crowell C, Kittoe R, Bredell H, Machaba M, Willamson C, Janssens W, Jallow S, van der Groen G, Shao Y, Jacob M, Samuel NM, de Rivera IL, Ngo-Giang-Huong N, Cassol S, Alemnji G, Frenkel LM |
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| Jazyk: | angličtina |
| Zdroj: | Journal of acquired immune deficiency syndromes (1999) [J Acquir Immune Defic Syndr] 2008 Aug 01; Vol. 48 (4), pp. 418-27. |
| DOI: | 10.1097/QAI.0b013e31817ed7d7 |
| Abstrakt: | 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% (2,410) of 2,604 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. |
| Databáze: | MEDLINE |
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