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Development of a TaqMan-based multiplex real-time PCR for simultaneous detection of four feline diarrhea-associated viruses

Since their recent discovery, the prevalence of novel feline enteric viruses, including feline bocavirus 1 (FBoV-1), feline astrovirus (FeAstV), and feline kobuvirus (FeKoV), has been reported in China. Co-infections of these viruses with feline parvovirus (FPV) are common causes of diarrhea in cats...

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Autores principales: Zou, Junwei, Yu, Ju, Mu, Yuanyuan, Xie, Xiangyu, Wang, Run, Wu, Haiqiang, Liu, Xuan, Xu, Fazhi, Wang, Juhua, Wang, Yong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9669448/
https://www.ncbi.nlm.nih.gov/pubmed/36406081
http://dx.doi.org/10.3389/fvets.2022.1005759
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author Zou, Junwei
Yu, Ju
Mu, Yuanyuan
Xie, Xiangyu
Wang, Run
Wu, Haiqiang
Liu, Xuan
Xu, Fazhi
Wang, Juhua
Wang, Yong
author_facet Zou, Junwei
Yu, Ju
Mu, Yuanyuan
Xie, Xiangyu
Wang, Run
Wu, Haiqiang
Liu, Xuan
Xu, Fazhi
Wang, Juhua
Wang, Yong
author_sort Zou, Junwei
collection PubMed
description Since their recent discovery, the prevalence of novel feline enteric viruses, including feline bocavirus 1 (FBoV-1), feline astrovirus (FeAstV), and feline kobuvirus (FeKoV), has been reported in China. Co-infections of these viruses with feline parvovirus (FPV) are common causes of diarrhea in cats. Viral co-infections are difficult to identify because of their non-specific clinical signs. To detect and identify these viruses, a quick and specific pathogen-testing approach is required. Here, we establish a real-time PCR (qPCR) based on multiple TaqMan probes for the simultaneous detection of FBoV-1, FeAstV, FeKoV, and FPV. Specific primers and TaqMan fluorescent probes were designed to ensure specificity. The results showed that the detection limit of single qPCR was up to 10 copies, and the detection limit of multiplex qPCR was up to 100 copies, with correlation coefficients >0.995 in all cases. Clinical sample detection revealed a 25.19% (34/135) total rate of co-infection among the viruses and a 1.48% (2/135) quadruple infection rate. Thus, this multiplex qPCR approach can serve as a quick, sensitive, and specific diagnostic tool for FBoV-1, FeAstV, FeKoV, and FPV identification, and it may be utilized for routine surveillance of these emerging and reemerging feline enteric viruses.
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spelling pubmed-96694482022-11-18 Development of a TaqMan-based multiplex real-time PCR for simultaneous detection of four feline diarrhea-associated viruses Zou, Junwei Yu, Ju Mu, Yuanyuan Xie, Xiangyu Wang, Run Wu, Haiqiang Liu, Xuan Xu, Fazhi Wang, Juhua Wang, Yong Front Vet Sci Veterinary Science Since their recent discovery, the prevalence of novel feline enteric viruses, including feline bocavirus 1 (FBoV-1), feline astrovirus (FeAstV), and feline kobuvirus (FeKoV), has been reported in China. Co-infections of these viruses with feline parvovirus (FPV) are common causes of diarrhea in cats. Viral co-infections are difficult to identify because of their non-specific clinical signs. To detect and identify these viruses, a quick and specific pathogen-testing approach is required. Here, we establish a real-time PCR (qPCR) based on multiple TaqMan probes for the simultaneous detection of FBoV-1, FeAstV, FeKoV, and FPV. Specific primers and TaqMan fluorescent probes were designed to ensure specificity. The results showed that the detection limit of single qPCR was up to 10 copies, and the detection limit of multiplex qPCR was up to 100 copies, with correlation coefficients >0.995 in all cases. Clinical sample detection revealed a 25.19% (34/135) total rate of co-infection among the viruses and a 1.48% (2/135) quadruple infection rate. Thus, this multiplex qPCR approach can serve as a quick, sensitive, and specific diagnostic tool for FBoV-1, FeAstV, FeKoV, and FPV identification, and it may be utilized for routine surveillance of these emerging and reemerging feline enteric viruses. Frontiers Media S.A. 2022-11-03 /pmc/articles/PMC9669448/ /pubmed/36406081 http://dx.doi.org/10.3389/fvets.2022.1005759 Text en Copyright © 2022 Zou, Yu, Mu, Xie, Wang, Wu, Liu, Xu, Wang and Wang. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Veterinary Science
Zou, Junwei
Yu, Ju
Mu, Yuanyuan
Xie, Xiangyu
Wang, Run
Wu, Haiqiang
Liu, Xuan
Xu, Fazhi
Wang, Juhua
Wang, Yong
Development of a TaqMan-based multiplex real-time PCR for simultaneous detection of four feline diarrhea-associated viruses
title Development of a TaqMan-based multiplex real-time PCR for simultaneous detection of four feline diarrhea-associated viruses
title_full Development of a TaqMan-based multiplex real-time PCR for simultaneous detection of four feline diarrhea-associated viruses
title_fullStr Development of a TaqMan-based multiplex real-time PCR for simultaneous detection of four feline diarrhea-associated viruses
title_full_unstemmed Development of a TaqMan-based multiplex real-time PCR for simultaneous detection of four feline diarrhea-associated viruses
title_short Development of a TaqMan-based multiplex real-time PCR for simultaneous detection of four feline diarrhea-associated viruses
title_sort development of a taqman-based multiplex real-time pcr for simultaneous detection of four feline diarrhea-associated viruses
topic Veterinary Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9669448/
https://www.ncbi.nlm.nih.gov/pubmed/36406081
http://dx.doi.org/10.3389/fvets.2022.1005759
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