Cargando…

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-...

Descripción completa

Detalles Bibliográficos
Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
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
_version_ 1784737299528941568
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
work_keys_str_mv AT wangsuchun establishmentandapplicationofaquadruplerealtimertpcrfordetectingavianmetapneumovirus
AT jiangnan establishmentandapplicationofaquadruplerealtimertpcrfordetectingavianmetapneumovirus
AT jianglijian establishmentandapplicationofaquadruplerealtimertpcrfordetectingavianmetapneumovirus
AT zhuangqingye establishmentandapplicationofaquadruplerealtimertpcrfordetectingavianmetapneumovirus
AT chenqiong establishmentandapplicationofaquadruplerealtimertpcrfordetectingavianmetapneumovirus
AT houguangyu establishmentandapplicationofaquadruplerealtimertpcrfordetectingavianmetapneumovirus
AT xiaozhiyu establishmentandapplicationofaquadruplerealtimertpcrfordetectingavianmetapneumovirus
AT zhaoran establishmentandapplicationofaquadruplerealtimertpcrfordetectingavianmetapneumovirus
AT liyang establishmentandapplicationofaquadruplerealtimertpcrfordetectingavianmetapneumovirus
AT zhaochenglong establishmentandapplicationofaquadruplerealtimertpcrfordetectingavianmetapneumovirus
AT zhangfuyou establishmentandapplicationofaquadruplerealtimertpcrfordetectingavianmetapneumovirus
AT yujianmin establishmentandapplicationofaquadruplerealtimertpcrfordetectingavianmetapneumovirus
AT lijinping establishmentandapplicationofaquadruplerealtimertpcrfordetectingavianmetapneumovirus
AT liuhualei establishmentandapplicationofaquadruplerealtimertpcrfordetectingavianmetapneumovirus
AT sunfuliang establishmentandapplicationofaquadruplerealtimertpcrfordetectingavianmetapneumovirus
AT wangkaicheng establishmentandapplicationofaquadruplerealtimertpcrfordetectingavianmetapneumovirus