Cargando…
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...
Autores principales: | , , , , , , , , , |
---|---|
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 |
_version_ | 1783339221726003200 |
---|---|
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. |
format | Online Article Text |
id | pubmed-6042793 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT katodaisuke aneasyrapidandsensitivemethodfordetectionofdrugresistantinfluenzavirusbyusingasialidasefluorescentimagingprobebtp3neu5ac AT kurebayashiyuuki aneasyrapidandsensitivemethodfordetectionofdrugresistantinfluenzavirusbyusingasialidasefluorescentimagingprobebtp3neu5ac AT takahashitadanobu aneasyrapidandsensitivemethodfordetectionofdrugresistantinfluenzavirusbyusingasialidasefluorescentimagingprobebtp3neu5ac AT otsubotadamune aneasyrapidandsensitivemethodfordetectionofdrugresistantinfluenzavirusbyusingasialidasefluorescentimagingprobebtp3neu5ac AT otakehitomi aneasyrapidandsensitivemethodfordetectionofdrugresistantinfluenzavirusbyusingasialidasefluorescentimagingprobebtp3neu5ac AT yamazakimika aneasyrapidandsensitivemethodfordetectionofdrugresistantinfluenzavirusbyusingasialidasefluorescentimagingprobebtp3neu5ac AT tamotochihiro aneasyrapidandsensitivemethodfordetectionofdrugresistantinfluenzavirusbyusingasialidasefluorescentimagingprobebtp3neu5ac AT minamiakira aneasyrapidandsensitivemethodfordetectionofdrugresistantinfluenzavirusbyusingasialidasefluorescentimagingprobebtp3neu5ac AT ikedakiyoshi aneasyrapidandsensitivemethodfordetectionofdrugresistantinfluenzavirusbyusingasialidasefluorescentimagingprobebtp3neu5ac AT suzukitakashi aneasyrapidandsensitivemethodfordetectionofdrugresistantinfluenzavirusbyusingasialidasefluorescentimagingprobebtp3neu5ac AT katodaisuke easyrapidandsensitivemethodfordetectionofdrugresistantinfluenzavirusbyusingasialidasefluorescentimagingprobebtp3neu5ac AT kurebayashiyuuki easyrapidandsensitivemethodfordetectionofdrugresistantinfluenzavirusbyusingasialidasefluorescentimagingprobebtp3neu5ac AT takahashitadanobu easyrapidandsensitivemethodfordetectionofdrugresistantinfluenzavirusbyusingasialidasefluorescentimagingprobebtp3neu5ac AT otsubotadamune easyrapidandsensitivemethodfordetectionofdrugresistantinfluenzavirusbyusingasialidasefluorescentimagingprobebtp3neu5ac AT otakehitomi easyrapidandsensitivemethodfordetectionofdrugresistantinfluenzavirusbyusingasialidasefluorescentimagingprobebtp3neu5ac AT yamazakimika easyrapidandsensitivemethodfordetectionofdrugresistantinfluenzavirusbyusingasialidasefluorescentimagingprobebtp3neu5ac AT tamotochihiro easyrapidandsensitivemethodfordetectionofdrugresistantinfluenzavirusbyusingasialidasefluorescentimagingprobebtp3neu5ac AT minamiakira easyrapidandsensitivemethodfordetectionofdrugresistantinfluenzavirusbyusingasialidasefluorescentimagingprobebtp3neu5ac AT ikedakiyoshi easyrapidandsensitivemethodfordetectionofdrugresistantinfluenzavirusbyusingasialidasefluorescentimagingprobebtp3neu5ac AT suzukitakashi easyrapidandsensitivemethodfordetectionofdrugresistantinfluenzavirusbyusingasialidasefluorescentimagingprobebtp3neu5ac |