文献类型：期刊文献

英文题名：Loop-mediated isothermal amplification method for rapid detection of Vibrio alginolyticus, the causative agent of vibriosis in mariculture fish

作者：Cai, S. H.[1,2];Lu, Y. S.[1,2];Wu, Z. -H.[1,2];Jian, J. C.[1,2];Wang, B.[1,2];Huang, Y. C.[1,2]

机构：[1]Guangdong Ocean Univ, Coll Fisheries, Zhanjiang 524025, Guangdong, Peoples R China;[2]Guangdong Prov Key Lab Pathogen Biol & Epidemiol, Zhanjiang, Peoples R China

年份：2010

卷号：50

期号：5

起止页码：480

外文期刊名：LETTERS IN APPLIED MICROBIOLOGY

收录：SCI-EXPANDED(收录号：WOS:000276406400007)、、Scopus(收录号：2-s2.0-77950481239)、WOS

基金：This work was supported by research grants from Oceanic and Fisheries Administrator of Guangdong Province in the People's Republic of China.

语种：英文

外文关键词：loop-mediated isothermal amplification; PCR; rapid detection; Vibrio alginolyticus

外文摘要：Aims: The purpose of this study was to develop a loop-mediated isothermal amplification (LAMP) method for the rapid, sensitive and simple detection of Vibrio alginolyticus in mariculture fish. Methods and Results: LAMP primers were designed by targeting the gyrB gene. With Bst DNA polymerase, the target DNA can be clearly amplified for 60 min at 64 degrees C in a simple water bath. The detection sensitivity of the LAMP assay for the detection of V. alginolyticus is about 3 center dot 7 x 102 CFU ml-1 (3 center dot 7 CFU per reaction). LAMP products could be judged with agar gel or naked eye after the addition of SYBR Green I. There were no cross-reactions with other bacterial strains indicating a high specificity of the LAMP. The LAMP method was applied to detect V. alginolyticus-infected fish tissues effectively. Conclusions: The LAMP established in this study is a simple, sensitive, specific, inexpensive and rapid protocol for the detection of V. alginolyticus. Significance and Impact of the Study: This LAMP method provides an important diagnostic tool for the detection of V. alginolyticus infection both in the laboratory and field.