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Rapid and sensitive detection of Senecavirus A by reverse transcription loop-mediated isothermal amplification combined with a lateral flow dipstick method

Senecavirus A (SVA) is a critical pathogen causing vesicular lesions in sows and acute death of newborn piglets, resulting in very large economic losses in the pig industry. To restrict the transmission of SVA, an establishment of an effective diagnostic method is crucial for the prevention and cont...

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Autores principales: Li, Jinhui, Liang, Weifang, Xu, Shuaifei, Shi, Jian, Zhou, Xia, Liu, Bowen, Yu, Li, Xiong, Jingfeng, Si, Guangbin, He, Dongsheng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6497277/
https://www.ncbi.nlm.nih.gov/pubmed/31048910
http://dx.doi.org/10.1371/journal.pone.0216245
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author Li, Jinhui
Liang, Weifang
Xu, Shuaifei
Shi, Jian
Zhou, Xia
Liu, Bowen
Yu, Li
Xiong, Jingfeng
Si, Guangbin
He, Dongsheng
author_facet Li, Jinhui
Liang, Weifang
Xu, Shuaifei
Shi, Jian
Zhou, Xia
Liu, Bowen
Yu, Li
Xiong, Jingfeng
Si, Guangbin
He, Dongsheng
author_sort Li, Jinhui
collection PubMed
description Senecavirus A (SVA) is a critical pathogen causing vesicular lesions in sows and acute death of newborn piglets, resulting in very large economic losses in the pig industry. To restrict the transmission of SVA, an establishment of an effective diagnostic method is crucial for the prevention and control of the disease. However, traditional detection methods often have many drawbacks. In this study, reverse transcription loop-mediated isothermal amplification (RT-LAMP) was combined with a lateral flow dipstick (LFD) to detect SVA. The resulting RT-LAMP-LFD assay was performed at 60°C for 50 min and then directly judged on an LFD visualization strip. This method shows high specificity and sensitivity to SVA. The detection limit of RT-LAMP was 4.56x10(-8) ng/μL RNA, approximately 11 copies/μL RNA, and it was 10 times more sensitive than RT-PCR. This detection method’s positive rate for clinical samples is comparable to that of RT-PCR. This method is time saving and highly efficient and is thus expected to be used to diagnose SVA infections in this field.
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spelling pubmed-64972772019-05-17 Rapid and sensitive detection of Senecavirus A by reverse transcription loop-mediated isothermal amplification combined with a lateral flow dipstick method Li, Jinhui Liang, Weifang Xu, Shuaifei Shi, Jian Zhou, Xia Liu, Bowen Yu, Li Xiong, Jingfeng Si, Guangbin He, Dongsheng PLoS One Research Article Senecavirus A (SVA) is a critical pathogen causing vesicular lesions in sows and acute death of newborn piglets, resulting in very large economic losses in the pig industry. To restrict the transmission of SVA, an establishment of an effective diagnostic method is crucial for the prevention and control of the disease. However, traditional detection methods often have many drawbacks. In this study, reverse transcription loop-mediated isothermal amplification (RT-LAMP) was combined with a lateral flow dipstick (LFD) to detect SVA. The resulting RT-LAMP-LFD assay was performed at 60°C for 50 min and then directly judged on an LFD visualization strip. This method shows high specificity and sensitivity to SVA. The detection limit of RT-LAMP was 4.56x10(-8) ng/μL RNA, approximately 11 copies/μL RNA, and it was 10 times more sensitive than RT-PCR. This detection method’s positive rate for clinical samples is comparable to that of RT-PCR. This method is time saving and highly efficient and is thus expected to be used to diagnose SVA infections in this field. Public Library of Science 2019-05-02 /pmc/articles/PMC6497277/ /pubmed/31048910 http://dx.doi.org/10.1371/journal.pone.0216245 Text en © 2019 Li et al 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 author and source are credited.
spellingShingle Research Article
Li, Jinhui
Liang, Weifang
Xu, Shuaifei
Shi, Jian
Zhou, Xia
Liu, Bowen
Yu, Li
Xiong, Jingfeng
Si, Guangbin
He, Dongsheng
Rapid and sensitive detection of Senecavirus A by reverse transcription loop-mediated isothermal amplification combined with a lateral flow dipstick method
title Rapid and sensitive detection of Senecavirus A by reverse transcription loop-mediated isothermal amplification combined with a lateral flow dipstick method
title_full Rapid and sensitive detection of Senecavirus A by reverse transcription loop-mediated isothermal amplification combined with a lateral flow dipstick method
title_fullStr Rapid and sensitive detection of Senecavirus A by reverse transcription loop-mediated isothermal amplification combined with a lateral flow dipstick method
title_full_unstemmed Rapid and sensitive detection of Senecavirus A by reverse transcription loop-mediated isothermal amplification combined with a lateral flow dipstick method
title_short Rapid and sensitive detection of Senecavirus A by reverse transcription loop-mediated isothermal amplification combined with a lateral flow dipstick method
title_sort rapid and sensitive detection of senecavirus a by reverse transcription loop-mediated isothermal amplification combined with a lateral flow dipstick method
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6497277/
https://www.ncbi.nlm.nih.gov/pubmed/31048910
http://dx.doi.org/10.1371/journal.pone.0216245
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