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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...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2022
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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. |
format | Online Article Text |
id | pubmed-9669448 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
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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