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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...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Hindawi
2020
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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. |
format | Online Article Text |
id | pubmed-7199591 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Hindawi |
record_format | MEDLINE/PubMed |
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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