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Establishment and application of a quadruple real-time RT-PCR for detecting avian metapneumovirus
In order to develop an appropriate method for high-throughput detection of avian metapneumovirus, a quadruple real-time reverse-transcription polymerase chain reaction assay was established with four pairs of specific primers and four specific probes based on the G or M gene of aMPV-A, aMPV-B, aMPV-...
Autores principales: | , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9239461/ https://www.ncbi.nlm.nih.gov/pubmed/35763505 http://dx.doi.org/10.1371/journal.pone.0270708 |
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author | Wang, Suchun Jiang, Nan Jiang, Lijian Zhuang, Qingye Chen, Qiong Hou, Guangyu Xiao, Zhiyu Zhao, Ran Li, Yang Zhao, Chenglong Zhang, Fuyou Yu, Jianmin Li, Jinping Liu, Hualei Sun, Fuliang Wang, Kaicheng |
author_facet | Wang, Suchun Jiang, Nan Jiang, Lijian Zhuang, Qingye Chen, Qiong Hou, Guangyu Xiao, Zhiyu Zhao, Ran Li, Yang Zhao, Chenglong Zhang, Fuyou Yu, Jianmin Li, Jinping Liu, Hualei Sun, Fuliang Wang, Kaicheng |
author_sort | Wang, Suchun |
collection | PubMed |
description | In order to develop an appropriate method for high-throughput detection of avian metapneumovirus, a quadruple real-time reverse-transcription polymerase chain reaction assay was established with four pairs of specific primers and four specific probes based on the G or M gene of aMPV-A, aMPV-B, aMPV-C and aMPV-D. Its specificity and sensitivity were evaluated, and clinical samples were tested by the method. The results showed that all the four subgroups of avian metapneumovirus can be detected in the quadruple real-time RT-PCR assay simultaneously, with a detection limit of 100–1000 cRNA copies/reaction. The other common poultry viruses were negative. In the avian clinical sample detection, 39 out of 1920 clinical samples collected from 8 provinces were positive. Compared with published RT-PCR assays, the κ value of the quadruple real-time RT-PCR assay in 1920 avian clinical samples was 1.000 (P < 0.001). The established method could be used for the rapid detection of the four subgroups of avian metapneumovirus with high specificity and high sensitivity. |
format | Online Article Text |
id | pubmed-9239461 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-92394612022-06-29 Establishment and application of a quadruple real-time RT-PCR for detecting avian metapneumovirus Wang, Suchun Jiang, Nan Jiang, Lijian Zhuang, Qingye Chen, Qiong Hou, Guangyu Xiao, Zhiyu Zhao, Ran Li, Yang Zhao, Chenglong Zhang, Fuyou Yu, Jianmin Li, Jinping Liu, Hualei Sun, Fuliang Wang, Kaicheng PLoS One Research Article In order to develop an appropriate method for high-throughput detection of avian metapneumovirus, a quadruple real-time reverse-transcription polymerase chain reaction assay was established with four pairs of specific primers and four specific probes based on the G or M gene of aMPV-A, aMPV-B, aMPV-C and aMPV-D. Its specificity and sensitivity were evaluated, and clinical samples were tested by the method. The results showed that all the four subgroups of avian metapneumovirus can be detected in the quadruple real-time RT-PCR assay simultaneously, with a detection limit of 100–1000 cRNA copies/reaction. The other common poultry viruses were negative. In the avian clinical sample detection, 39 out of 1920 clinical samples collected from 8 provinces were positive. Compared with published RT-PCR assays, the κ value of the quadruple real-time RT-PCR assay in 1920 avian clinical samples was 1.000 (P < 0.001). The established method could be used for the rapid detection of the four subgroups of avian metapneumovirus with high specificity and high sensitivity. Public Library of Science 2022-06-28 /pmc/articles/PMC9239461/ /pubmed/35763505 http://dx.doi.org/10.1371/journal.pone.0270708 Text en © 2022 Wang et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://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 Wang, Suchun Jiang, Nan Jiang, Lijian Zhuang, Qingye Chen, Qiong Hou, Guangyu Xiao, Zhiyu Zhao, Ran Li, Yang Zhao, Chenglong Zhang, Fuyou Yu, Jianmin Li, Jinping Liu, Hualei Sun, Fuliang Wang, Kaicheng Establishment and application of a quadruple real-time RT-PCR for detecting avian metapneumovirus |
title | Establishment and application of a quadruple real-time RT-PCR for detecting avian metapneumovirus |
title_full | Establishment and application of a quadruple real-time RT-PCR for detecting avian metapneumovirus |
title_fullStr | Establishment and application of a quadruple real-time RT-PCR for detecting avian metapneumovirus |
title_full_unstemmed | Establishment and application of a quadruple real-time RT-PCR for detecting avian metapneumovirus |
title_short | Establishment and application of a quadruple real-time RT-PCR for detecting avian metapneumovirus |
title_sort | establishment and application of a quadruple real-time rt-pcr for detecting avian metapneumovirus |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9239461/ https://www.ncbi.nlm.nih.gov/pubmed/35763505 http://dx.doi.org/10.1371/journal.pone.0270708 |
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