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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...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2014
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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. |
format | Online Article Text |
id | pubmed-4026823 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
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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