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Development of a TaqMan Based Real-Time Fluorescent Quantitative PCR Assay for Detection of Porcine Cytomegalovirus in Semen

This study described a TaqMan based real-time fluorescent quantitative PCR (qPCR) method to detect porcine cytomegalovirus (PCMV) infection, targeting the conserved region of the DNA polymerase (DPOL) gene. The standard curve showed a linear regression relationship with a coefficient of 0.999 and a...

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Detalles Bibliográficos
Autores principales: Chen, Rujing, Chen, Qiuyong, Wu, Xuemin, Che, Yongliang, Wang, Chenyan, Wang, Longbai, Yan, Shan, Zhou, Lunjiang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7199591/
https://www.ncbi.nlm.nih.gov/pubmed/32420350
http://dx.doi.org/10.1155/2020/5673145
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author Chen, Rujing
Chen, Qiuyong
Wu, Xuemin
Che, Yongliang
Wang, Chenyan
Wang, Longbai
Yan, Shan
Zhou, Lunjiang
author_facet Chen, Rujing
Chen, Qiuyong
Wu, Xuemin
Che, Yongliang
Wang, Chenyan
Wang, Longbai
Yan, Shan
Zhou, Lunjiang
author_sort Chen, Rujing
collection PubMed
description This study described a TaqMan based real-time fluorescent quantitative PCR (qPCR) method to detect porcine cytomegalovirus (PCMV) infection, targeting the conserved region of the DNA polymerase (DPOL) gene. The standard curve showed a linear regression relationship with a coefficient of 0.999 and a slope of y = −3.249x + 38.958 corresponding to the amplification efficiency at 99.8%. The limit of the qPCR method was 51.9 copies/μl. The established qPCR method showed excellent specificity, with no cross-reaction observed with common porcine pathogens. The coefficient of variation for intra-assay and interassay variability ranged up to 1.51% and 2.24%, respectively. PCMV positive signals can be found in semen using this qPCR method, which suggested that we should pay more attention to PCMV contamination in semen in order to eliminate PCMV infection in artificial insemination and xenotransplantation.
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spelling pubmed-71995912020-05-15 Development of a TaqMan Based Real-Time Fluorescent Quantitative PCR Assay for Detection of Porcine Cytomegalovirus in Semen Chen, Rujing Chen, Qiuyong Wu, Xuemin Che, Yongliang Wang, Chenyan Wang, Longbai Yan, Shan Zhou, Lunjiang Biomed Res Int Research Article This study described a TaqMan based real-time fluorescent quantitative PCR (qPCR) method to detect porcine cytomegalovirus (PCMV) infection, targeting the conserved region of the DNA polymerase (DPOL) gene. The standard curve showed a linear regression relationship with a coefficient of 0.999 and a slope of y = −3.249x + 38.958 corresponding to the amplification efficiency at 99.8%. The limit of the qPCR method was 51.9 copies/μl. The established qPCR method showed excellent specificity, with no cross-reaction observed with common porcine pathogens. The coefficient of variation for intra-assay and interassay variability ranged up to 1.51% and 2.24%, respectively. PCMV positive signals can be found in semen using this qPCR method, which suggested that we should pay more attention to PCMV contamination in semen in order to eliminate PCMV infection in artificial insemination and xenotransplantation. Hindawi 2020-01-10 /pmc/articles/PMC7199591/ /pubmed/32420350 http://dx.doi.org/10.1155/2020/5673145 Text en Copyright © 2020 Rujing Chen et al. http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Chen, Rujing
Chen, Qiuyong
Wu, Xuemin
Che, Yongliang
Wang, Chenyan
Wang, Longbai
Yan, Shan
Zhou, Lunjiang
Development of a TaqMan Based Real-Time Fluorescent Quantitative PCR Assay for Detection of Porcine Cytomegalovirus in Semen
title Development of a TaqMan Based Real-Time Fluorescent Quantitative PCR Assay for Detection of Porcine Cytomegalovirus in Semen
title_full Development of a TaqMan Based Real-Time Fluorescent Quantitative PCR Assay for Detection of Porcine Cytomegalovirus in Semen
title_fullStr Development of a TaqMan Based Real-Time Fluorescent Quantitative PCR Assay for Detection of Porcine Cytomegalovirus in Semen
title_full_unstemmed Development of a TaqMan Based Real-Time Fluorescent Quantitative PCR Assay for Detection of Porcine Cytomegalovirus in Semen
title_short Development of a TaqMan Based Real-Time Fluorescent Quantitative PCR Assay for Detection of Porcine Cytomegalovirus in Semen
title_sort development of a taqman based real-time fluorescent quantitative pcr assay for detection of porcine cytomegalovirus in semen
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7199591/
https://www.ncbi.nlm.nih.gov/pubmed/32420350
http://dx.doi.org/10.1155/2020/5673145
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