Fluorometric assay for phenotypic differentiation of drug-resistant HIV mutants

Convenient drug-resistance testing of viral mutants is indispensable to effective treatment of viral infection. We developed a novel fluorometric assay for phenotypic differentiation of drug-resistant mutants of human immunodeficiency virus-I protease (HIV-PR) which uses enzymatic and peptide-specif...

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Autores principales: Zhu, Qinchang, Yu, Zhiqiang, Kabashima, Tsutomu, Yin, Sheng, Dragusha, Shpend, El-Mahdy, Ahmed F. M., Ejupi, Valon, Shibata, Takayuki, Kai, Masaaki
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2015
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4437315/
https://www.ncbi.nlm.nih.gov/pubmed/25988960
http://dx.doi.org/10.1038/srep10323
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author Zhu, Qinchang
Yu, Zhiqiang
Kabashima, Tsutomu
Yin, Sheng
Dragusha, Shpend
El-Mahdy, Ahmed F. M.
Ejupi, Valon
Shibata, Takayuki
Kai, Masaaki
author_facet Zhu, Qinchang
Yu, Zhiqiang
Kabashima, Tsutomu
Yin, Sheng
Dragusha, Shpend
El-Mahdy, Ahmed F. M.
Ejupi, Valon
Shibata, Takayuki
Kai, Masaaki
author_sort Zhu, Qinchang
collection PubMed
description Convenient drug-resistance testing of viral mutants is indispensable to effective treatment of viral infection. We developed a novel fluorometric assay for phenotypic differentiation of drug-resistant mutants of human immunodeficiency virus-I protease (HIV-PR) which uses enzymatic and peptide-specific fluorescence (FL) reactions and high-performance liquid chromatography (HPLC) of three HIV-PR substrates. This assay protocol enables use of non-purified enzyme sources and multiple substrates for the enzymatic reaction. In this study, susceptibility of HIV mutations to drugs was evaluated by selective formation of three FL products after the enzymatic HIV-PR reaction. This proof-of-concept study indicates that the present HPLC-FL method could be an alternative to current phenotypic assays for the evaluation of HIV drug resistance.
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spelling pubmed-44373152015-06-01 Fluorometric assay for phenotypic differentiation of drug-resistant HIV mutants Zhu, Qinchang Yu, Zhiqiang Kabashima, Tsutomu Yin, Sheng Dragusha, Shpend El-Mahdy, Ahmed F. M. Ejupi, Valon Shibata, Takayuki Kai, Masaaki Sci Rep Article Convenient drug-resistance testing of viral mutants is indispensable to effective treatment of viral infection. We developed a novel fluorometric assay for phenotypic differentiation of drug-resistant mutants of human immunodeficiency virus-I protease (HIV-PR) which uses enzymatic and peptide-specific fluorescence (FL) reactions and high-performance liquid chromatography (HPLC) of three HIV-PR substrates. This assay protocol enables use of non-purified enzyme sources and multiple substrates for the enzymatic reaction. In this study, susceptibility of HIV mutations to drugs was evaluated by selective formation of three FL products after the enzymatic HIV-PR reaction. This proof-of-concept study indicates that the present HPLC-FL method could be an alternative to current phenotypic assays for the evaluation of HIV drug resistance. Nature Publishing Group 2015-05-19 /pmc/articles/PMC4437315/ /pubmed/25988960 http://dx.doi.org/10.1038/srep10323 Text en Copyright © 2015, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Zhu, Qinchang
Yu, Zhiqiang
Kabashima, Tsutomu
Yin, Sheng
Dragusha, Shpend
El-Mahdy, Ahmed F. M.
Ejupi, Valon
Shibata, Takayuki
Kai, Masaaki
Fluorometric assay for phenotypic differentiation of drug-resistant HIV mutants
title Fluorometric assay for phenotypic differentiation of drug-resistant HIV mutants
title_full Fluorometric assay for phenotypic differentiation of drug-resistant HIV mutants
title_fullStr Fluorometric assay for phenotypic differentiation of drug-resistant HIV mutants
title_full_unstemmed Fluorometric assay for phenotypic differentiation of drug-resistant HIV mutants
title_short Fluorometric assay for phenotypic differentiation of drug-resistant HIV mutants
title_sort fluorometric assay for phenotypic differentiation of drug-resistant hiv mutants
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4437315/
https://www.ncbi.nlm.nih.gov/pubmed/25988960
http://dx.doi.org/10.1038/srep10323
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