Rapid, sensitive detection of Vibrio anguillarum using loop-mediated isothermal amplification

doi:10.1007/s00343-010-9247-3

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	Rapid, sensitive detection of Vibrio anguillarum using loop-mediated isothermal amplification
	Gao Hongwei 1,2; Li Fuhua 1; Zhang Xiaojun 1; Wang Bing 1; Xiang Jianhai1
	2010
发表期刊	CHINESE JOURNAL OF OCEANOLOGY AND LIMNOLOGY
卷号	28 期号:1 页码:62-66
文章类型	Article
摘要	Vibrio anguillarum is an important bacterial pathogen of aquatic organisms and a significant problem in aquatic farming. The rapid detection and identification of V. anguillarum, and other pathogens that infect marine organisms, is crucial to effective disease management. In this study, we developed a loop-mediated amplification (LAMP) assay to detect V. anguillarum in an hour in a single tube without the need for thermal cycling. Conserved regions of the metalloproteinase (empA) gene of V. anguillarum served as the targets for primer design. A fragment of the empA gene was amplified at 65 degrees C in the presence of the primer mixture and Bst DNA polymerase. In the optimized LAMP assay, 6.7 pg of V. anguillarum DNA could be detected. Six strains of V. anguillarum and 17 strains of non-V. anguillarum bacteria were used in this study to evaluate the species specificity of the primers. The six V. anguillarum strains gave a positive result in the LAMP assay. This method was also validated in V. anguillarum-infected fish. This LAMP method is more sensitive than PCR in the detection of V. anguillarum and shows good species specificity. The LAMP assay is therefore an effective method for the quick detection of V. anguillarum both in the laboratory and in the field.
关键词	Loop-mediated Isothermal Amplification (Lamp) Detection Assay Empa Gene Vibrio Anguillarum
DOI	10.1007/s00343-010-9247-3
收录类别	SCI
语种	英语
WOS记录号	WOS:000275654100008
引用统计	被引频次：5[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/32827
专题	实验海洋生物学重点实验室
作者单位	1.Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China 2.Shandong Entry Exit Inspect & Quarantine Bur Peop, Qingdao 266001, Peoples R China
第一作者单位	中国科学院海洋研究所
推荐引用方式 GB/T 7714	Gao Hongwei,Li Fuhua,Zhang Xiaojun,et al. Rapid, sensitive detection of Vibrio anguillarum using loop-mediated isothermal amplification[J]. CHINESE JOURNAL OF OCEANOLOGY AND LIMNOLOGY,2010,28(1):62-66.
APA	Gao Hongwei,Li Fuhua,Zhang Xiaojun,Wang Bing,&Xiang Jianhai.(2010).Rapid, sensitive detection of Vibrio anguillarum using loop-mediated isothermal amplification.CHINESE JOURNAL OF OCEANOLOGY AND LIMNOLOGY,28(1),62-66.
MLA	Gao Hongwei,et al."Rapid, sensitive detection of Vibrio anguillarum using loop-mediated isothermal amplification".CHINESE JOURNAL OF OCEANOLOGY AND LIMNOLOGY 28.1(2010):62-66.

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