Cargando…

RT‐LAMP for rapid diagnosis of coronavirus SARS‐CoV‐2

The pandemic coronavirus SARS‐CoV‐2 in the world has caused a large infected population suffering from COVID‐19. To curb the spreading of the virus, WHO urgently demanded an extension of screening and testing; thus, a rapid and simple diagnostic method is needed. We applied a reverse transcription‐l...

Descripción completa

Detalles Bibliográficos
Autores principales: Huang, Wei E., Lim, Boon, Hsu, Chia‐Chen, Xiong, Dan, Wu, Wei, Yu, Yejiong, Jia, Huidong, Wang, Yun, Zeng, Yida, Ji, Mengmeng, Chang, Hong, Zhang, Xiuming, Wang, Hui, Cui, Zhanfeng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7264870/
https://www.ncbi.nlm.nih.gov/pubmed/32333644
http://dx.doi.org/10.1111/1751-7915.13586
_version_ 1783541018952466432
author Huang, Wei E.
Lim, Boon
Hsu, Chia‐Chen
Xiong, Dan
Wu, Wei
Yu, Yejiong
Jia, Huidong
Wang, Yun
Zeng, Yida
Ji, Mengmeng
Chang, Hong
Zhang, Xiuming
Wang, Hui
Cui, Zhanfeng
author_facet Huang, Wei E.
Lim, Boon
Hsu, Chia‐Chen
Xiong, Dan
Wu, Wei
Yu, Yejiong
Jia, Huidong
Wang, Yun
Zeng, Yida
Ji, Mengmeng
Chang, Hong
Zhang, Xiuming
Wang, Hui
Cui, Zhanfeng
author_sort Huang, Wei E.
collection PubMed
description The pandemic coronavirus SARS‐CoV‐2 in the world has caused a large infected population suffering from COVID‐19. To curb the spreading of the virus, WHO urgently demanded an extension of screening and testing; thus, a rapid and simple diagnostic method is needed. We applied a reverse transcription‐loop‐mediated isothermal amplification (RT‐LAMP) to achieve the detection of SARS‐CoV‐2 in 30 min. We designed four sets of LAMP primers (6 primers in each set), targeting the viral RNA of SARS‐CoV‐2 in the regions of orf1ab, S gene and N gene. A colorimetric change was used to report the results, which enables the outcome of viral RNA amplification to be read by the naked eye without the need of expensive or dedicated instrument. The sensitivity can be 80 copies of viral RNA per ml in a sample. We validated the RT‐LAMP method in a hospital in China, employing 16 clinic samples with 8 positives and 8 negatives. The testing results are consistent with the conventional RT‐qPCR. In addition, we also show that one‐step process without RNA extraction is feasible to achieve RNA amplification directly from a sample. This rapid, simple and sensitive RT‐LAMP method paves a way for a large screening at public domain and hospitals, particularly regional hospitals and medical centres in rural areas.
format Online
Article
Text
id pubmed-7264870
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher John Wiley and Sons Inc.
record_format MEDLINE/PubMed
spelling pubmed-72648702020-06-03 RT‐LAMP for rapid diagnosis of coronavirus SARS‐CoV‐2 Huang, Wei E. Lim, Boon Hsu, Chia‐Chen Xiong, Dan Wu, Wei Yu, Yejiong Jia, Huidong Wang, Yun Zeng, Yida Ji, Mengmeng Chang, Hong Zhang, Xiuming Wang, Hui Cui, Zhanfeng Microb Biotechnol Research Articles The pandemic coronavirus SARS‐CoV‐2 in the world has caused a large infected population suffering from COVID‐19. To curb the spreading of the virus, WHO urgently demanded an extension of screening and testing; thus, a rapid and simple diagnostic method is needed. We applied a reverse transcription‐loop‐mediated isothermal amplification (RT‐LAMP) to achieve the detection of SARS‐CoV‐2 in 30 min. We designed four sets of LAMP primers (6 primers in each set), targeting the viral RNA of SARS‐CoV‐2 in the regions of orf1ab, S gene and N gene. A colorimetric change was used to report the results, which enables the outcome of viral RNA amplification to be read by the naked eye without the need of expensive or dedicated instrument. The sensitivity can be 80 copies of viral RNA per ml in a sample. We validated the RT‐LAMP method in a hospital in China, employing 16 clinic samples with 8 positives and 8 negatives. The testing results are consistent with the conventional RT‐qPCR. In addition, we also show that one‐step process without RNA extraction is feasible to achieve RNA amplification directly from a sample. This rapid, simple and sensitive RT‐LAMP method paves a way for a large screening at public domain and hospitals, particularly regional hospitals and medical centres in rural areas. John Wiley and Sons Inc. 2020-04-25 /pmc/articles/PMC7264870/ /pubmed/32333644 http://dx.doi.org/10.1111/1751-7915.13586 Text en © 2020 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Articles
Huang, Wei E.
Lim, Boon
Hsu, Chia‐Chen
Xiong, Dan
Wu, Wei
Yu, Yejiong
Jia, Huidong
Wang, Yun
Zeng, Yida
Ji, Mengmeng
Chang, Hong
Zhang, Xiuming
Wang, Hui
Cui, Zhanfeng
RT‐LAMP for rapid diagnosis of coronavirus SARS‐CoV‐2
title RT‐LAMP for rapid diagnosis of coronavirus SARS‐CoV‐2
title_full RT‐LAMP for rapid diagnosis of coronavirus SARS‐CoV‐2
title_fullStr RT‐LAMP for rapid diagnosis of coronavirus SARS‐CoV‐2
title_full_unstemmed RT‐LAMP for rapid diagnosis of coronavirus SARS‐CoV‐2
title_short RT‐LAMP for rapid diagnosis of coronavirus SARS‐CoV‐2
title_sort rt‐lamp for rapid diagnosis of coronavirus sars‐cov‐2
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7264870/
https://www.ncbi.nlm.nih.gov/pubmed/32333644
http://dx.doi.org/10.1111/1751-7915.13586
work_keys_str_mv AT huangweie rtlampforrapiddiagnosisofcoronavirussarscov2
AT limboon rtlampforrapiddiagnosisofcoronavirussarscov2
AT hsuchiachen rtlampforrapiddiagnosisofcoronavirussarscov2
AT xiongdan rtlampforrapiddiagnosisofcoronavirussarscov2
AT wuwei rtlampforrapiddiagnosisofcoronavirussarscov2
AT yuyejiong rtlampforrapiddiagnosisofcoronavirussarscov2
AT jiahuidong rtlampforrapiddiagnosisofcoronavirussarscov2
AT wangyun rtlampforrapiddiagnosisofcoronavirussarscov2
AT zengyida rtlampforrapiddiagnosisofcoronavirussarscov2
AT jimengmeng rtlampforrapiddiagnosisofcoronavirussarscov2
AT changhong rtlampforrapiddiagnosisofcoronavirussarscov2
AT zhangxiuming rtlampforrapiddiagnosisofcoronavirussarscov2
AT wanghui rtlampforrapiddiagnosisofcoronavirussarscov2
AT cuizhanfeng rtlampforrapiddiagnosisofcoronavirussarscov2