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Rapid, sensitive detection of Vibrio anguillarum using loop-mediated isothermal amplification
Gao Hongwei1,2; Li Fuhua1; Zhang Xiaojun1; Wang Bing1; 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
DOI10.1007/s00343-010-9247-3
收录类别SCI
语种英语
WOS记录号WOS:000275654100008
引用统计
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
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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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