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Development of a duplex real-time RT-PCR for the simultaneous detection and differentiation of Theiler’s murine encephalomyelitis virus and rat theilovirus

Theiler’s murine encephalomyelitis virus (TMEV) and rat theilovirus (RTV), the member of the genus Cardiovirus, are widespread in laboratory mice and rats, and are potential contaminants of biological materials. Cardioviruses infection may cause serious complications in biomedical research. To impro...

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Autores principales: Yuan, Wen, Wang, Jing, Xu, Fengjiao, Huang, Bihong, Lian, Yuexiao, Rao, Dan, Yin, Xueqin, Wu, Miaoli, Zhu, Yujun, Zhang, Yu, Huang, Ren, Guo, Pengju
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier B.V. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7119687/
https://www.ncbi.nlm.nih.gov/pubmed/27396678
http://dx.doi.org/10.1016/j.jviromet.2016.07.004
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author Yuan, Wen
Wang, Jing
Xu, Fengjiao
Huang, Bihong
Lian, Yuexiao
Rao, Dan
Yin, Xueqin
Wu, Miaoli
Zhu, Yujun
Zhang, Yu
Huang, Ren
Guo, Pengju
author_facet Yuan, Wen
Wang, Jing
Xu, Fengjiao
Huang, Bihong
Lian, Yuexiao
Rao, Dan
Yin, Xueqin
Wu, Miaoli
Zhu, Yujun
Zhang, Yu
Huang, Ren
Guo, Pengju
author_sort Yuan, Wen
collection PubMed
description Theiler’s murine encephalomyelitis virus (TMEV) and rat theilovirus (RTV), the member of the genus Cardiovirus, are widespread in laboratory mice and rats, and are potential contaminants of biological materials. Cardioviruses infection may cause serious complications in biomedical research. To improve the efficiency of routine screening for Cardioviruses infection, a duplex real-time reverse transcriptase polymerase chain reaction (RT-PCR) assay was developed for simultaneous detection and differentiation of TMEV and RTV. The duplex assay was specific for reference strains of TMEV and RTV, and no cross-reaction was found with seven other rodent viruses. The limits of detection of both TMEV and RTV were 4 × 10(1) copies RNA/reaction. Reproducibility was estimated using standard dilutions, with coefficients of variation <3.1%. 439 clinical samples were evaluated by both duplex real-time RT-PCR and conventional RT-PCR. For 439 clinical samples,95 samples were positive for TMEV and 72 samples were positive for RTV using duplex real-time RT-PCR approach, whereas only 77 samples were positive for TMEV and 66 samples were positive for RTV when conventional RT-PCR was applied. Mixed infections were found in 20 samples when analyzed by conventional RT-PCR whereas 30 samples were found to be mixed infection when duplex real-time RT-PCR was applied. This duplex assay provides a useful tool for routine health monitoring and screening of contaminated biological materials of these two viruses.
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spelling pubmed-71196872020-04-08 Development of a duplex real-time RT-PCR for the simultaneous detection and differentiation of Theiler’s murine encephalomyelitis virus and rat theilovirus Yuan, Wen Wang, Jing Xu, Fengjiao Huang, Bihong Lian, Yuexiao Rao, Dan Yin, Xueqin Wu, Miaoli Zhu, Yujun Zhang, Yu Huang, Ren Guo, Pengju J Virol Methods Article Theiler’s murine encephalomyelitis virus (TMEV) and rat theilovirus (RTV), the member of the genus Cardiovirus, are widespread in laboratory mice and rats, and are potential contaminants of biological materials. Cardioviruses infection may cause serious complications in biomedical research. To improve the efficiency of routine screening for Cardioviruses infection, a duplex real-time reverse transcriptase polymerase chain reaction (RT-PCR) assay was developed for simultaneous detection and differentiation of TMEV and RTV. The duplex assay was specific for reference strains of TMEV and RTV, and no cross-reaction was found with seven other rodent viruses. The limits of detection of both TMEV and RTV were 4 × 10(1) copies RNA/reaction. Reproducibility was estimated using standard dilutions, with coefficients of variation <3.1%. 439 clinical samples were evaluated by both duplex real-time RT-PCR and conventional RT-PCR. For 439 clinical samples,95 samples were positive for TMEV and 72 samples were positive for RTV using duplex real-time RT-PCR approach, whereas only 77 samples were positive for TMEV and 66 samples were positive for RTV when conventional RT-PCR was applied. Mixed infections were found in 20 samples when analyzed by conventional RT-PCR whereas 30 samples were found to be mixed infection when duplex real-time RT-PCR was applied. This duplex assay provides a useful tool for routine health monitoring and screening of contaminated biological materials of these two viruses. Elsevier B.V. 2016-10 2016-07-07 /pmc/articles/PMC7119687/ /pubmed/27396678 http://dx.doi.org/10.1016/j.jviromet.2016.07.004 Text en © 2016 Elsevier B.V. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.
spellingShingle Article
Yuan, Wen
Wang, Jing
Xu, Fengjiao
Huang, Bihong
Lian, Yuexiao
Rao, Dan
Yin, Xueqin
Wu, Miaoli
Zhu, Yujun
Zhang, Yu
Huang, Ren
Guo, Pengju
Development of a duplex real-time RT-PCR for the simultaneous detection and differentiation of Theiler’s murine encephalomyelitis virus and rat theilovirus
title Development of a duplex real-time RT-PCR for the simultaneous detection and differentiation of Theiler’s murine encephalomyelitis virus and rat theilovirus
title_full Development of a duplex real-time RT-PCR for the simultaneous detection and differentiation of Theiler’s murine encephalomyelitis virus and rat theilovirus
title_fullStr Development of a duplex real-time RT-PCR for the simultaneous detection and differentiation of Theiler’s murine encephalomyelitis virus and rat theilovirus
title_full_unstemmed Development of a duplex real-time RT-PCR for the simultaneous detection and differentiation of Theiler’s murine encephalomyelitis virus and rat theilovirus
title_short Development of a duplex real-time RT-PCR for the simultaneous detection and differentiation of Theiler’s murine encephalomyelitis virus and rat theilovirus
title_sort development of a duplex real-time rt-pcr for the simultaneous detection and differentiation of theiler’s murine encephalomyelitis virus and rat theilovirus
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7119687/
https://www.ncbi.nlm.nih.gov/pubmed/27396678
http://dx.doi.org/10.1016/j.jviromet.2016.07.004
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