WarmStart colorimetric loop-mediated isothermal amplification for the one-tube, contamination-free and visualization detection of Shigella flexneri

Objectives: Shigella flexneri (S. flexneri) is prevalent worldwide and the most common Shigella in many countries, causing highly contagious diarrhea, which seriously threatens public health. This study aimed to develop a colorimetric loop-mediated isothermal amplification (LAMP) for the rapid, accu...

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Autores principales:	Yaoqiang Shi, Min Xu, Xiaoqiong Duan, Shilin Li, Jia-wei Ding, Limin Chen
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Loop-mediated isothermal amplification Shigella flexneri Hypothetical protein gene Colorimetric detection Point-of-care testing Infectious and parasitic diseases RC109-216
Acceso en línea:	https://doaj.org/article/a30283f9f45940b7bc8b1b0102fc70ab
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spelling	oai:doaj.org-article:a30283f9f45940b7bc8b1b0102fc70ab2021-11-30T04:14:10ZWarmStart colorimetric loop-mediated isothermal amplification for the one-tube, contamination-free and visualization detection of Shigella flexneri1201-971210.1016/j.ijid.2021.09.015https://doaj.org/article/a30283f9f45940b7bc8b1b0102fc70ab2021-11-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S1201971221007219https://doaj.org/toc/1201-9712Objectives: Shigella flexneri (S. flexneri) is prevalent worldwide and the most common Shigella in many countries, causing highly contagious diarrhea, which seriously threatens public health. This study aimed to develop a colorimetric loop-mediated isothermal amplification (LAMP) for the rapid, accurate, and visualization detection of S. flexneri. Methods: According to the screened specific genes of S. flexneri, three groups of LAMP primers were designed and evaluated, and the colorimetric LAMP reaction volume was optimized. The specificity of the colorimetric LAMP was validated by 20 S. flexneri and 96 non-S. flexneri clinical isolates. In addition, the sensitivity of the developed assay was evaluated by the serial 10-fold dilutions of plasmid DNA. Results: A colorimetric LAMP assay was developed based on the specific S. flexneri hypothetical protein gene (Accession: AE014073 Region: 4170556.4171068). The colorimetric LAMP method had good specificity for detecting S. flexneri and enabled detection of S. flexneri within 30 minutes, with a plasmid detection limit of 7*10° copies/μL. The results of amplification could be easily identified by color. Conclusions: This colorimetric LAMP assay could be used for rapid and accurate diagnosis of S. flexneri infection, especially in remote hospitals and laboratories with under-equipped medical facilities, and in situations where an urgent diagnosis is needed.Yaoqiang ShiMin XuXiaoqiong DuanShilin LiJia-wei DingLimin ChenElsevierarticleLoop-mediated isothermal amplificationShigella flexneriHypothetical protein geneColorimetric detectionPoint-of-care testingInfectious and parasitic diseasesRC109-216ENInternational Journal of Infectious Diseases, Vol 112, Iss , Pp 55-62 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Loop-mediated isothermal amplification Shigella flexneri Hypothetical protein gene Colorimetric detection Point-of-care testing Infectious and parasitic diseases RC109-216
spellingShingle	Loop-mediated isothermal amplification Shigella flexneri Hypothetical protein gene Colorimetric detection Point-of-care testing Infectious and parasitic diseases RC109-216 Yaoqiang Shi Min Xu Xiaoqiong Duan Shilin Li Jia-wei Ding Limin Chen WarmStart colorimetric loop-mediated isothermal amplification for the one-tube, contamination-free and visualization detection of Shigella flexneri
description	Objectives: Shigella flexneri (S. flexneri) is prevalent worldwide and the most common Shigella in many countries, causing highly contagious diarrhea, which seriously threatens public health. This study aimed to develop a colorimetric loop-mediated isothermal amplification (LAMP) for the rapid, accurate, and visualization detection of S. flexneri. Methods: According to the screened specific genes of S. flexneri, three groups of LAMP primers were designed and evaluated, and the colorimetric LAMP reaction volume was optimized. The specificity of the colorimetric LAMP was validated by 20 S. flexneri and 96 non-S. flexneri clinical isolates. In addition, the sensitivity of the developed assay was evaluated by the serial 10-fold dilutions of plasmid DNA. Results: A colorimetric LAMP assay was developed based on the specific S. flexneri hypothetical protein gene (Accession: AE014073 Region: 4170556.4171068). The colorimetric LAMP method had good specificity for detecting S. flexneri and enabled detection of S. flexneri within 30 minutes, with a plasmid detection limit of 7*10° copies/μL. The results of amplification could be easily identified by color. Conclusions: This colorimetric LAMP assay could be used for rapid and accurate diagnosis of S. flexneri infection, especially in remote hospitals and laboratories with under-equipped medical facilities, and in situations where an urgent diagnosis is needed.
format	article
author	Yaoqiang Shi Min Xu Xiaoqiong Duan Shilin Li Jia-wei Ding Limin Chen
author_facet	Yaoqiang Shi Min Xu Xiaoqiong Duan Shilin Li Jia-wei Ding Limin Chen
author_sort	Yaoqiang Shi
title	WarmStart colorimetric loop-mediated isothermal amplification for the one-tube, contamination-free and visualization detection of Shigella flexneri
title_short	WarmStart colorimetric loop-mediated isothermal amplification for the one-tube, contamination-free and visualization detection of Shigella flexneri
title_full	WarmStart colorimetric loop-mediated isothermal amplification for the one-tube, contamination-free and visualization detection of Shigella flexneri
title_fullStr	WarmStart colorimetric loop-mediated isothermal amplification for the one-tube, contamination-free and visualization detection of Shigella flexneri
title_full_unstemmed	WarmStart colorimetric loop-mediated isothermal amplification for the one-tube, contamination-free and visualization detection of Shigella flexneri
title_sort	warmstart colorimetric loop-mediated isothermal amplification for the one-tube, contamination-free and visualization detection of shigella flexneri
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/a30283f9f45940b7bc8b1b0102fc70ab
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_version_	1718406823643447296

WarmStart colorimetric loop-mediated isothermal amplification for the one-tube, contamination-free and visualization detection of Shigella flexneri

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