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A triplex real-time PCR method to detect African swine fever virus gene-deleted and wild type strains

Currently there is still no effective vaccines and drugs available for African swine fever virus (ASFV), a life-threatening virus to domestic pigs and wild boars. Therefore, accurate diagnosis is important for the prevention and control of the virus. In this study, we developed a triplex real-time P...

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Autores principales: Yang, Hao, Peng, Zhong, Song, Wenbo, Zhang, Chen, Fan, Jie, Chen, Hongjian, Hua, Lin, Pei, Jie, Tang, Xibiao, Chen, Huanchun, Wu, Bin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9521421/
https://www.ncbi.nlm.nih.gov/pubmed/36187818
http://dx.doi.org/10.3389/fvets.2022.943099
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author Yang, Hao
Peng, Zhong
Song, Wenbo
Zhang, Chen
Fan, Jie
Chen, Hongjian
Hua, Lin
Pei, Jie
Tang, Xibiao
Chen, Huanchun
Wu, Bin
author_facet Yang, Hao
Peng, Zhong
Song, Wenbo
Zhang, Chen
Fan, Jie
Chen, Hongjian
Hua, Lin
Pei, Jie
Tang, Xibiao
Chen, Huanchun
Wu, Bin
author_sort Yang, Hao
collection PubMed
description Currently there is still no effective vaccines and drugs available for African swine fever virus (ASFV), a life-threatening virus to domestic pigs and wild boars. Therefore, accurate diagnosis is important for the prevention and control of the virus. In this study, we developed a triplex real-time PCR method to detect and differentiate ASFV gene-deleted and wild type strains based on three viral genes B646L, MGF_360-14L gene, and CD2v. Standard curves plotted showed that there was a strong linear correlation (R(2) > 0.99) between Ct values and the corresponding copy numbers of synthesized standard plasmids. The detection limits of the method for B646L, MGF_360-14L, and CD2v were 78.9, 47.0, and 82.1 copies/μl, respectively. Detection results of different types of swine viruses showed that the method only gave amplification curves to ASFV. Finally, we found the triplex real-time PCR method developed in this study displayed better results on detecting the laboratory sample mocks, and it could be used as a supplemental method to detect ASFV genotype I strains. These findings suggest that the triplex real-time PCR method developed in this study have good specificity and sensitivity. This triplex real-time PCR method might also represent an effective tool for the detection of ASFV gene-deleted and wild type strains.
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spelling pubmed-95214212022-09-30 A triplex real-time PCR method to detect African swine fever virus gene-deleted and wild type strains Yang, Hao Peng, Zhong Song, Wenbo Zhang, Chen Fan, Jie Chen, Hongjian Hua, Lin Pei, Jie Tang, Xibiao Chen, Huanchun Wu, Bin Front Vet Sci Veterinary Science Currently there is still no effective vaccines and drugs available for African swine fever virus (ASFV), a life-threatening virus to domestic pigs and wild boars. Therefore, accurate diagnosis is important for the prevention and control of the virus. In this study, we developed a triplex real-time PCR method to detect and differentiate ASFV gene-deleted and wild type strains based on three viral genes B646L, MGF_360-14L gene, and CD2v. Standard curves plotted showed that there was a strong linear correlation (R(2) > 0.99) between Ct values and the corresponding copy numbers of synthesized standard plasmids. The detection limits of the method for B646L, MGF_360-14L, and CD2v were 78.9, 47.0, and 82.1 copies/μl, respectively. Detection results of different types of swine viruses showed that the method only gave amplification curves to ASFV. Finally, we found the triplex real-time PCR method developed in this study displayed better results on detecting the laboratory sample mocks, and it could be used as a supplemental method to detect ASFV genotype I strains. These findings suggest that the triplex real-time PCR method developed in this study have good specificity and sensitivity. This triplex real-time PCR method might also represent an effective tool for the detection of ASFV gene-deleted and wild type strains. Frontiers Media S.A. 2022-09-15 /pmc/articles/PMC9521421/ /pubmed/36187818 http://dx.doi.org/10.3389/fvets.2022.943099 Text en Copyright © 2022 Yang, Peng, Song, Zhang, Fan, Chen, Hua, Pei, Tang, Chen and Wu. 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
Yang, Hao
Peng, Zhong
Song, Wenbo
Zhang, Chen
Fan, Jie
Chen, Hongjian
Hua, Lin
Pei, Jie
Tang, Xibiao
Chen, Huanchun
Wu, Bin
A triplex real-time PCR method to detect African swine fever virus gene-deleted and wild type strains
title A triplex real-time PCR method to detect African swine fever virus gene-deleted and wild type strains
title_full A triplex real-time PCR method to detect African swine fever virus gene-deleted and wild type strains
title_fullStr A triplex real-time PCR method to detect African swine fever virus gene-deleted and wild type strains
title_full_unstemmed A triplex real-time PCR method to detect African swine fever virus gene-deleted and wild type strains
title_short A triplex real-time PCR method to detect African swine fever virus gene-deleted and wild type strains
title_sort triplex real-time pcr method to detect african swine fever virus gene-deleted and wild type strains
topic Veterinary Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9521421/
https://www.ncbi.nlm.nih.gov/pubmed/36187818
http://dx.doi.org/10.3389/fvets.2022.943099
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