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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...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2019
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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. |
format | Online Article Text |
id | pubmed-6497277 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
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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