环介导恒温扩增法快速检测海产品中的副溶血弧菌  

Rapid Detection of Vibrio parahemolyticus in Seafood by Loop-mediated Isothermal Amplification

作　　者：彭帅[1] 石磊[2] 

机构地区：[1]华南理工大学生物科学与工程学院,广州510640 [2]华南理工大学轻工与食品学院,广州510640

出　　处：《生物技术通报》2011年第2期184-186,191,共4页Biotechnology Bulletin

基　　金：国家自然科学基金项目(20877028);2008年珠海市产学研专项项目(PC20082034)

摘　　要：副溶血弧菌广泛分布于海水或海产品中,人类摄入或接触污染的水源和食物易引起感染。近年来,有关致病性弧菌引起腹泻的报道逐渐增多,但GB标准的细菌学诊断方法检测周期长达1周左右,而且操作较为复杂,难以满足控制疾病暴发和传播的需要,就这一现状,建立了一套不仅快速、准确,而且操作简便、不依赖昂贵仪器的检测方法,应用于海产品中副溶血弧菌快速检测。采用环介导恒温扩增方法(LAMP),针对副溶血弧菌的gyrB基因设计特异引物,进行恒温扩增。使用该方法最低检出限达到101CFU/mL,灵敏度可以达到0.1pg副溶血弧菌基因组DNA,为海产品中副溶血弧菌的检测提供了一个新的辅助方法。Vibrio parahemolyticus is widely distributed in seawater or seafood,human contact with contaminated water or food will be easily caused infection.Recently,reports of diarrhea caused by the pathogenic Vibrio gradually increased.But the GB standard bacteriological diagnostics test cycle up to 1 week or so,and the operation is complex,difficult to meet the need of control of disease outbreaks and spread.Loop-mediated isothermal amplification is a novel nucleic acid amplification method in which reagents react under isothermal conditions with high specificity,efficiency,and rapidity.In this study,the LAMP method was employed in rapid detection of Vibrio parahemolyticus in seafood.The species-specific primers were designed by targeting the gyrB gene.The results showed that the method is sensitive;the 101CFU/mL of Vibrio parahemolyticus or 0.1 pg genome DNA can be detected within 4 hours.This new method might facilitate genetic analysis applied in Vibrio parahemolyticus in seafood.

关 键 词：副溶血弧菌 环介导恒温扩增(LAMP) gyrB基因 

分 类 号：TS254.7[轻工技术与工程—水产品加工及贮藏工程]