Fisheries College, Guangdong Ocean University, Zhanjiang, China.
Lett Appl Microbiol. 2010 May;50(5):480-5. doi: 10.1111/j.1472-765X.2010.02823.x. Epub 2010 Feb 11.
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.
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 3.7 x 10(2) CFU ml(-1) (3.7 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.
The LAMP established in this study is a simple, sensitive, specific, inexpensive and rapid protocol for the detection of V. alginolyticus.
This LAMP method provides an important diagnostic tool for the detection of V. alginolyticus infection both in the laboratory and field.
本研究旨在开发一种环介导等温扩增（LAMP）方法，用于快速、敏感、简便地检测水产养殖鱼类中的弧菌。
针对 gyrB 基因设计了 LAMP 引物。在 Bst DNA 聚合酶的作用下，目标 DNA 可在 64°C 的简单水浴中于 60 分钟内被清晰扩增。LAMP 检测法检测 V. alginolyticus 的检测灵敏度约为 3.7 x 10(2) CFU ml(-1)（每个反应 3.7 CFU）。加入 SYBR Green I 后，可通过琼脂凝胶或肉眼判断 LAMP 产物。与其他细菌菌株无交叉反应，表明具有高度特异性。该 LAMP 方法可有效用于检测感染弧菌的鱼类组织。
本研究建立的 LAMP 是一种简单、敏感、特异、廉价和快速的检测 V. alginolyticus 的方法。
该 LAMP 方法为实验室和现场检测弧菌感染提供了重要的诊断工具。
