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An easy, rapid, and sensitive method for detection of drug-resistant influenza virus by using a sialidase fluorescent imaging probe, BTP3-Neu5Ac

Immunochromatographic kits and RT-PCR are widely used as diagnostic tools for influenza detection in clinical and hygiene fields. Immunochromatographic kits are useful for differential typing of influenza A and influenza B but cannot show if the detected virus strains have acquired drug resistance a...

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Autores principales: Kato, Daisuke, Kurebayashi, Yuuki, Takahashi, Tadanobu, Otsubo, Tadamune, Otake, Hitomi, Yamazaki, Mika, Tamoto, Chihiro, Minami, Akira, Ikeda, Kiyoshi, Suzuki, Takashi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6042793/
https://www.ncbi.nlm.nih.gov/pubmed/30001430
http://dx.doi.org/10.1371/journal.pone.0200761
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author Kato, Daisuke
Kurebayashi, Yuuki
Takahashi, Tadanobu
Otsubo, Tadamune
Otake, Hitomi
Yamazaki, Mika
Tamoto, Chihiro
Minami, Akira
Ikeda, Kiyoshi
Suzuki, Takashi
author_facet Kato, Daisuke
Kurebayashi, Yuuki
Takahashi, Tadanobu
Otsubo, Tadamune
Otake, Hitomi
Yamazaki, Mika
Tamoto, Chihiro
Minami, Akira
Ikeda, Kiyoshi
Suzuki, Takashi
author_sort Kato, Daisuke
collection PubMed
description Immunochromatographic kits and RT-PCR are widely used as diagnostic tools for influenza detection in clinical and hygiene fields. Immunochromatographic kits are useful for differential typing of influenza A and influenza B but cannot show if the detected virus strains have acquired drug resistance against neuraminidase inhibitors that target sialidase activity of viral neuraminidase. Although RT-PCR enables determination of drug-resistant mutants, its efficacy is limited to viruses carrying a known substitution in their neuraminidase genome sequence. In the present study, an easy, rapid and sensitive method for detection of drug-resistant influenza viruses regardless of major antigenic changes or genomic mutations was developed. By using the method in combination with virus-concentrated membranes in centrifugal filter units and a sialidase imaging probe, 2-(benzothiazol-2-yl)-4-bromophenyl-N-acetylneuraminic acid (BTP3-Neu5Ac), sialidase activity of influenza neuraminidase was visualized on membranes by the green fluorescence of produced hydrophobic BTP3 under UV irradiation with a handheld UV flashlight. Fluorescence images in the presence or absence of neuraminidase inhibitors clearly discriminated drug-resistant influenza viruses from drug-sensitive ones. The assay can be done within 15 min. The detection sensitivity was shown to be equal to or higher than the sensitivities of commercial immunochromatographic kits. The assay will be a powerful tool for screening and monitoring of emerging drug-resistant influenza viruses and would help clinicians decide effective antiviral treatment strategies when such mutants have become prevalent.
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spelling pubmed-60427932018-07-26 An easy, rapid, and sensitive method for detection of drug-resistant influenza virus by using a sialidase fluorescent imaging probe, BTP3-Neu5Ac Kato, Daisuke Kurebayashi, Yuuki Takahashi, Tadanobu Otsubo, Tadamune Otake, Hitomi Yamazaki, Mika Tamoto, Chihiro Minami, Akira Ikeda, Kiyoshi Suzuki, Takashi PLoS One Research Article Immunochromatographic kits and RT-PCR are widely used as diagnostic tools for influenza detection in clinical and hygiene fields. Immunochromatographic kits are useful for differential typing of influenza A and influenza B but cannot show if the detected virus strains have acquired drug resistance against neuraminidase inhibitors that target sialidase activity of viral neuraminidase. Although RT-PCR enables determination of drug-resistant mutants, its efficacy is limited to viruses carrying a known substitution in their neuraminidase genome sequence. In the present study, an easy, rapid and sensitive method for detection of drug-resistant influenza viruses regardless of major antigenic changes or genomic mutations was developed. By using the method in combination with virus-concentrated membranes in centrifugal filter units and a sialidase imaging probe, 2-(benzothiazol-2-yl)-4-bromophenyl-N-acetylneuraminic acid (BTP3-Neu5Ac), sialidase activity of influenza neuraminidase was visualized on membranes by the green fluorescence of produced hydrophobic BTP3 under UV irradiation with a handheld UV flashlight. Fluorescence images in the presence or absence of neuraminidase inhibitors clearly discriminated drug-resistant influenza viruses from drug-sensitive ones. The assay can be done within 15 min. The detection sensitivity was shown to be equal to or higher than the sensitivities of commercial immunochromatographic kits. The assay will be a powerful tool for screening and monitoring of emerging drug-resistant influenza viruses and would help clinicians decide effective antiviral treatment strategies when such mutants have become prevalent. Public Library of Science 2018-07-12 /pmc/articles/PMC6042793/ /pubmed/30001430 http://dx.doi.org/10.1371/journal.pone.0200761 Text en © 2018 Kato et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Kato, Daisuke
Kurebayashi, Yuuki
Takahashi, Tadanobu
Otsubo, Tadamune
Otake, Hitomi
Yamazaki, Mika
Tamoto, Chihiro
Minami, Akira
Ikeda, Kiyoshi
Suzuki, Takashi
An easy, rapid, and sensitive method for detection of drug-resistant influenza virus by using a sialidase fluorescent imaging probe, BTP3-Neu5Ac
title An easy, rapid, and sensitive method for detection of drug-resistant influenza virus by using a sialidase fluorescent imaging probe, BTP3-Neu5Ac
title_full An easy, rapid, and sensitive method for detection of drug-resistant influenza virus by using a sialidase fluorescent imaging probe, BTP3-Neu5Ac
title_fullStr An easy, rapid, and sensitive method for detection of drug-resistant influenza virus by using a sialidase fluorescent imaging probe, BTP3-Neu5Ac
title_full_unstemmed An easy, rapid, and sensitive method for detection of drug-resistant influenza virus by using a sialidase fluorescent imaging probe, BTP3-Neu5Ac
title_short An easy, rapid, and sensitive method for detection of drug-resistant influenza virus by using a sialidase fluorescent imaging probe, BTP3-Neu5Ac
title_sort easy, rapid, and sensitive method for detection of drug-resistant influenza virus by using a sialidase fluorescent imaging probe, btp3-neu5ac
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6042793/
https://www.ncbi.nlm.nih.gov/pubmed/30001430
http://dx.doi.org/10.1371/journal.pone.0200761
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