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Rapid detection of porcine kobuvirus in feces by reverse transcription loop-mediated isothermal amplification

BACKGROUND: PKV is a new emerging pathogen detected in diarrhea pigs. At present, no more detection methods were reported except RT-PCR method. this study was to develop a fast diagnostic method based on the LAMP reaction for rapid detection of PKV nucleic acid in fecal samples. FINDINGS: Two pairs...

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Autores principales: Li, Changlong, Chen, Jianfei, Shi, Hongyan, Zhang, Xin, Shi, Da, Han, Xiao, Chi, Yanbin, Feng, Li
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4026823/
https://www.ncbi.nlm.nih.gov/pubmed/24755372
http://dx.doi.org/10.1186/1743-422X-11-73
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author Li, Changlong
Chen, Jianfei
Shi, Hongyan
Zhang, Xin
Shi, Da
Han, Xiao
Chi, Yanbin
Feng, Li
author_facet Li, Changlong
Chen, Jianfei
Shi, Hongyan
Zhang, Xin
Shi, Da
Han, Xiao
Chi, Yanbin
Feng, Li
author_sort Li, Changlong
collection PubMed
description BACKGROUND: PKV is a new emerging pathogen detected in diarrhea pigs. At present, no more detection methods were reported except RT-PCR method. this study was to develop a fast diagnostic method based on the LAMP reaction for rapid detection of PKV nucleic acid in fecal samples. FINDINGS: Two pairs of primers were designed to amplify the conservative 3D gene of PKV genome. The PKV RT-LAMP method possessed well specificity and had 100 times higher sensitivity than common reverse transcription PCR (RT-PCR), which could detect up to 10 RNA copies of the target gene. CONCLUSIONS: The results showed that the optimal reaction condition for RT-LAMP was achieved at 64°C for 50 min. Furthermore, the RT-LAMP procedure does not demand special equipment and is time-saving.
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spelling pubmed-40268232014-05-21 Rapid detection of porcine kobuvirus in feces by reverse transcription loop-mediated isothermal amplification Li, Changlong Chen, Jianfei Shi, Hongyan Zhang, Xin Shi, Da Han, Xiao Chi, Yanbin Feng, Li Virol J Short Report BACKGROUND: PKV is a new emerging pathogen detected in diarrhea pigs. At present, no more detection methods were reported except RT-PCR method. this study was to develop a fast diagnostic method based on the LAMP reaction for rapid detection of PKV nucleic acid in fecal samples. FINDINGS: Two pairs of primers were designed to amplify the conservative 3D gene of PKV genome. The PKV RT-LAMP method possessed well specificity and had 100 times higher sensitivity than common reverse transcription PCR (RT-PCR), which could detect up to 10 RNA copies of the target gene. CONCLUSIONS: The results showed that the optimal reaction condition for RT-LAMP was achieved at 64°C for 50 min. Furthermore, the RT-LAMP procedure does not demand special equipment and is time-saving. BioMed Central 2014-04-23 /pmc/articles/PMC4026823/ /pubmed/24755372 http://dx.doi.org/10.1186/1743-422X-11-73 Text en Copyright © 2014 Li et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/4.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Short Report
Li, Changlong
Chen, Jianfei
Shi, Hongyan
Zhang, Xin
Shi, Da
Han, Xiao
Chi, Yanbin
Feng, Li
Rapid detection of porcine kobuvirus in feces by reverse transcription loop-mediated isothermal amplification
title Rapid detection of porcine kobuvirus in feces by reverse transcription loop-mediated isothermal amplification
title_full Rapid detection of porcine kobuvirus in feces by reverse transcription loop-mediated isothermal amplification
title_fullStr Rapid detection of porcine kobuvirus in feces by reverse transcription loop-mediated isothermal amplification
title_full_unstemmed Rapid detection of porcine kobuvirus in feces by reverse transcription loop-mediated isothermal amplification
title_short Rapid detection of porcine kobuvirus in feces by reverse transcription loop-mediated isothermal amplification
title_sort rapid detection of porcine kobuvirus in feces by reverse transcription loop-mediated isothermal amplification
topic Short Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4026823/
https://www.ncbi.nlm.nih.gov/pubmed/24755372
http://dx.doi.org/10.1186/1743-422X-11-73
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