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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...

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Autores principales: ZHANG, Qingshan, WAN, Chunhe, LI, Chenxi, BAI, Xiaofei, LIU, Ming, LIU, Siguo, ZHANG, Yun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Japanese Society of Veterinary Science 2017
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.
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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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