Development of a droplet digital PCR assay to detect bovine alphaherpesvirus 1 in bovine semen

BACKGROUND: Infectious bovine rhinotracheitis (IBR) caused by bovine alphaherpesvirus 1 (BoHV-1) is one of the most important contagious diseases in bovine. This is one of the most common infectious disease of cattle. This has led to high economic losses in the cattle farming industry. BoHV-1 can po...

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Autores principales: Yu, Zhichao, Zhao, Zhiguo, Chen, Linjun, Yan, Han, Cui, Qiang, Ju, Xianghong, Yong, Yanhong, Liu, Xiaoxi, Ma, Xingbin, Zhang, Guanhua
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Research
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8976375/
https://www.ncbi.nlm.nih.gov/pubmed/35366879
http://dx.doi.org/10.1186/s12917-022-03235-2
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author Yu, Zhichao
Zhao, Zhiguo
Chen, Linjun
Yan, Han
Cui, Qiang
Ju, Xianghong
Yong, Yanhong
Liu, Xiaoxi
Ma, Xingbin
Zhang, Guanhua
author_facet Yu, Zhichao
Zhao, Zhiguo
Chen, Linjun
Yan, Han
Cui, Qiang
Ju, Xianghong
Yong, Yanhong
Liu, Xiaoxi
Ma, Xingbin
Zhang, Guanhua
author_sort Yu, Zhichao
collection PubMed
description BACKGROUND: Infectious bovine rhinotracheitis (IBR) caused by bovine alphaherpesvirus 1 (BoHV-1) is one of the most important contagious diseases in bovine. This is one of the most common infectious disease of cattle. This has led to high economic losses in the cattle farming industry. BoHV-1 can potentially be transmitted via semen during natural or artificial insemination (AI). Therefore, testing methods for the early diagnosis of BoHV-1 infection are urgently needed for international trade of ruminant semen. In this study, we developed a novel droplet digital PCR (ddPCR) assay for the detection of BoHV-1 DNA in semen samples. RESULTS: The ddPCR results showed that the detection limit was 4.45 copies per reaction with high reproducibility. The established method was highly specific for BoHV-1 and did not show cross-reactivity with specify the organisms (BTV, BVDV, Brucella, M . bovis). The results of clinical sample testing showed that the positivity rate of ddPCR (87.8%) was higher than that of qPCR (84.1%). CONCLUSIONS: The ddPCR assay showed good accuracy for mixed samples and could be a new added diagnostic tool for detecting BoHV-1. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12917-022-03235-2.
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spelling pubmed-89763752022-04-03 Development of a droplet digital PCR assay to detect bovine alphaherpesvirus 1 in bovine semen Yu, Zhichao Zhao, Zhiguo Chen, Linjun Yan, Han Cui, Qiang Ju, Xianghong Yong, Yanhong Liu, Xiaoxi Ma, Xingbin Zhang, Guanhua BMC Vet Res Research BACKGROUND: Infectious bovine rhinotracheitis (IBR) caused by bovine alphaherpesvirus 1 (BoHV-1) is one of the most important contagious diseases in bovine. This is one of the most common infectious disease of cattle. This has led to high economic losses in the cattle farming industry. BoHV-1 can potentially be transmitted via semen during natural or artificial insemination (AI). Therefore, testing methods for the early diagnosis of BoHV-1 infection are urgently needed for international trade of ruminant semen. In this study, we developed a novel droplet digital PCR (ddPCR) assay for the detection of BoHV-1 DNA in semen samples. RESULTS: The ddPCR results showed that the detection limit was 4.45 copies per reaction with high reproducibility. The established method was highly specific for BoHV-1 and did not show cross-reactivity with specify the organisms (BTV, BVDV, Brucella, M . bovis). The results of clinical sample testing showed that the positivity rate of ddPCR (87.8%) was higher than that of qPCR (84.1%). CONCLUSIONS: The ddPCR assay showed good accuracy for mixed samples and could be a new added diagnostic tool for detecting BoHV-1. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12917-022-03235-2. BioMed Central 2022-04-02 /pmc/articles/PMC8976375/ /pubmed/35366879 http://dx.doi.org/10.1186/s12917-022-03235-2 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Yu, Zhichao
Zhao, Zhiguo
Chen, Linjun
Yan, Han
Cui, Qiang
Ju, Xianghong
Yong, Yanhong
Liu, Xiaoxi
Ma, Xingbin
Zhang, Guanhua
Development of a droplet digital PCR assay to detect bovine alphaherpesvirus 1 in bovine semen
title Development of a droplet digital PCR assay to detect bovine alphaherpesvirus 1 in bovine semen
title_full Development of a droplet digital PCR assay to detect bovine alphaherpesvirus 1 in bovine semen
title_fullStr Development of a droplet digital PCR assay to detect bovine alphaherpesvirus 1 in bovine semen
title_full_unstemmed Development of a droplet digital PCR assay to detect bovine alphaherpesvirus 1 in bovine semen
title_short Development of a droplet digital PCR assay to detect bovine alphaherpesvirus 1 in bovine semen
title_sort development of a droplet digital pcr assay to detect bovine alphaherpesvirus 1 in bovine semen
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8976375/
https://www.ncbi.nlm.nih.gov/pubmed/35366879
http://dx.doi.org/10.1186/s12917-022-03235-2
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