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Evaluation of a quantitative real-time PCR for rapid detection of Riemerella Anatipestifer infection in birds
To establish an accurate, rapid, and a quantifiable method for the detection of Riemerella anatipestifer infection, a widespread infectious disease in birds, we developed a TaqMan-based real-time PCR assay by using DtxR gene-specific primers and a TaqMan probe. The standard curve established with a...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Japanese Society of Veterinary Science
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5745191/ https://www.ncbi.nlm.nih.gov/pubmed/28781328 http://dx.doi.org/10.1292/jvms.17-0227 |
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author | ZHANG, Qingshan WAN, Chunhe LI, Chenxi BAI, Xiaofei LIU, Ming LIU, Siguo ZHANG, Yun |
author_facet | ZHANG, Qingshan WAN, Chunhe LI, Chenxi BAI, Xiaofei LIU, Ming LIU, Siguo ZHANG, Yun |
author_sort | ZHANG, Qingshan |
collection | PubMed |
description | To establish an accurate, rapid, and a quantifiable method for the detection of Riemerella anatipestifer infection, a widespread infectious disease in birds, we developed a TaqMan-based real-time PCR assay by using DtxR gene-specific primers and a TaqMan probe. The standard curve established with a linear correlation (R(2)) of 0.998 and efficiency of 99% between the Ct value and the logarithm of the plasmid copy number. The reproducibility and specificity of the real-time PCR assay were confirmed by using plasmids containing DtxR genes or DNAs extracted from well-known bacteria or viruses causing duck diseases. The real-time PCR assay was 100 times more sensitive than the conventional PCR. The results reveal that the established real-time PCR assay might be a useful method for diagnosis and quantitative detection of Riemerella anatipestifer in birds. |
format | Online Article Text |
id | pubmed-5745191 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | The Japanese Society of Veterinary Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-57451912018-01-05 Evaluation of a quantitative real-time PCR for rapid detection of Riemerella Anatipestifer infection in birds ZHANG, Qingshan WAN, Chunhe LI, Chenxi BAI, Xiaofei LIU, Ming LIU, Siguo ZHANG, Yun J Vet Med Sci Avian Pathology To establish an accurate, rapid, and a quantifiable method for the detection of Riemerella anatipestifer infection, a widespread infectious disease in birds, we developed a TaqMan-based real-time PCR assay by using DtxR gene-specific primers and a TaqMan probe. The standard curve established with a linear correlation (R(2)) of 0.998 and efficiency of 99% between the Ct value and the logarithm of the plasmid copy number. The reproducibility and specificity of the real-time PCR assay were confirmed by using plasmids containing DtxR genes or DNAs extracted from well-known bacteria or viruses causing duck diseases. The real-time PCR assay was 100 times more sensitive than the conventional PCR. The results reveal that the established real-time PCR assay might be a useful method for diagnosis and quantitative detection of Riemerella anatipestifer in birds. The Japanese Society of Veterinary Science 2017-08-05 2017-12 /pmc/articles/PMC5745191/ /pubmed/28781328 http://dx.doi.org/10.1292/jvms.17-0227 Text en ©2017 The Japanese Society of Veterinary Science This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License. (CC-BY-NC-ND 4.0: https://creativecommons.org/licenses/by-nc-nd/4.0/) |
spellingShingle | Avian Pathology ZHANG, Qingshan WAN, Chunhe LI, Chenxi BAI, Xiaofei LIU, Ming LIU, Siguo ZHANG, Yun Evaluation of a quantitative real-time PCR for rapid detection of Riemerella Anatipestifer infection in birds |
title | Evaluation of a quantitative real-time PCR for rapid detection of
Riemerella Anatipestifer infection in birds |
title_full | Evaluation of a quantitative real-time PCR for rapid detection of
Riemerella Anatipestifer infection in birds |
title_fullStr | Evaluation of a quantitative real-time PCR for rapid detection of
Riemerella Anatipestifer infection in birds |
title_full_unstemmed | Evaluation of a quantitative real-time PCR for rapid detection of
Riemerella Anatipestifer infection in birds |
title_short | Evaluation of a quantitative real-time PCR for rapid detection of
Riemerella Anatipestifer infection in birds |
title_sort | evaluation of a quantitative real-time pcr for rapid detection of
riemerella anatipestifer infection in birds |
topic | Avian Pathology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5745191/ https://www.ncbi.nlm.nih.gov/pubmed/28781328 http://dx.doi.org/10.1292/jvms.17-0227 |
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