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Development and evaluation of a TaqMan MGB RT-PCR assay for detection of H5 and N8 subtype influenza virus

BACKGROUND: Highly pathogenic influenza A (H5N8) viruses have caused several worldwide outbreaks in birds and are of potential risk to humans. Thus, a specific, rapid and sensitive method for detection is urgently needed. METHODS: In the present study, TaqMan minor groove binder probes and multiplex...

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Autores principales: Yang, Fan, Xu, Lihua, Liu, Fumin, Yao, Hangping, Wu, Nanping, Wu, Haibo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7391517/
https://www.ncbi.nlm.nih.gov/pubmed/32727378
http://dx.doi.org/10.1186/s12879-020-05277-z
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author Yang, Fan
Xu, Lihua
Liu, Fumin
Yao, Hangping
Wu, Nanping
Wu, Haibo
author_facet Yang, Fan
Xu, Lihua
Liu, Fumin
Yao, Hangping
Wu, Nanping
Wu, Haibo
author_sort Yang, Fan
collection PubMed
description BACKGROUND: Highly pathogenic influenza A (H5N8) viruses have caused several worldwide outbreaks in birds and are of potential risk to humans. Thus, a specific, rapid and sensitive method for detection is urgently needed. METHODS: In the present study, TaqMan minor groove binder probes and multiplex real-time RT-PCR primers were designed to target the H5 hemagglutinin and N8 neuraminidase genes. A total of 38 strains of avian influenza viruses and other viruses were selected to test the performance of the assay. RESULTS: The results showed that only H5 and N8 avian influenza viruses yielded a positive signal, while all other subtypes avian influenza viruses and other viruses were negative. High specificity, repeatability, and sensitivity were achieved, with a detection limit of 10 copies per reaction. CONCLUSIONS: The developed assay could be a powerful tool for rapid detection of H5N8 influenza viruses in the future.
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spelling pubmed-73915172020-07-31 Development and evaluation of a TaqMan MGB RT-PCR assay for detection of H5 and N8 subtype influenza virus Yang, Fan Xu, Lihua Liu, Fumin Yao, Hangping Wu, Nanping Wu, Haibo BMC Infect Dis Research Article BACKGROUND: Highly pathogenic influenza A (H5N8) viruses have caused several worldwide outbreaks in birds and are of potential risk to humans. Thus, a specific, rapid and sensitive method for detection is urgently needed. METHODS: In the present study, TaqMan minor groove binder probes and multiplex real-time RT-PCR primers were designed to target the H5 hemagglutinin and N8 neuraminidase genes. A total of 38 strains of avian influenza viruses and other viruses were selected to test the performance of the assay. RESULTS: The results showed that only H5 and N8 avian influenza viruses yielded a positive signal, while all other subtypes avian influenza viruses and other viruses were negative. High specificity, repeatability, and sensitivity were achieved, with a detection limit of 10 copies per reaction. CONCLUSIONS: The developed assay could be a powerful tool for rapid detection of H5N8 influenza viruses in the future. BioMed Central 2020-07-29 /pmc/articles/PMC7391517/ /pubmed/32727378 http://dx.doi.org/10.1186/s12879-020-05277-z Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research Article
Yang, Fan
Xu, Lihua
Liu, Fumin
Yao, Hangping
Wu, Nanping
Wu, Haibo
Development and evaluation of a TaqMan MGB RT-PCR assay for detection of H5 and N8 subtype influenza virus
title Development and evaluation of a TaqMan MGB RT-PCR assay for detection of H5 and N8 subtype influenza virus
title_full Development and evaluation of a TaqMan MGB RT-PCR assay for detection of H5 and N8 subtype influenza virus
title_fullStr Development and evaluation of a TaqMan MGB RT-PCR assay for detection of H5 and N8 subtype influenza virus
title_full_unstemmed Development and evaluation of a TaqMan MGB RT-PCR assay for detection of H5 and N8 subtype influenza virus
title_short Development and evaluation of a TaqMan MGB RT-PCR assay for detection of H5 and N8 subtype influenza virus
title_sort development and evaluation of a taqman mgb rt-pcr assay for detection of h5 and n8 subtype influenza virus
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7391517/
https://www.ncbi.nlm.nih.gov/pubmed/32727378
http://dx.doi.org/10.1186/s12879-020-05277-z
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