环介导等温扩增技术快速检测椰毒假单胞菌的研究  

Rapid detection of Pseudomonas cocovenenans by loop-mediated isothermal amplification

作　　者：马晓燕[1] 张蕴哲[2] 王羽[1] 刘哲 王晓丽[4] 张伟[1] 

机构地区：[1]河北农业大学食品科技学院,河北保定071001 [2]河北大学附属医院,河北保定071000 [3]石家庄科技信息职业技术学院,河北石家庄050081 [4]保定科技职业学院,河北保定071000

出　　处：《食品工业科技》2013年第3期321-324,共4页Science and Technology of Food Industry

基　　金：河北省自然科学基金资助项目(C2008000216)

摘　　要：建立环介导等温扩增技术(LAMP)快速检测椰毒假单胞菌的方法。根据公布的椰毒假单胞菌16S～23SrRNA基因序列设计引物,建立了LAMP反应体系,在此基础上检测了蜡样芽孢杆菌菌液和人工污染蜡样芽孢杆菌的银耳样品,并将LAMP法与PCR法进行比较。结果表明,LAMP检测方法具有较高的特异性和敏感性,椰毒假单胞菌的检出限为5.4CFU/mL,是PCR方法检测灵敏度的1000倍,人工污染银耳样品中的椰毒假单胞菌的检出限为76CFU/g,样品中椰毒假单胞菌的检测过程(包括DNA提取、LAMP和电泳)可在2h内完成,因此LAMP可以简便快速有效地检测食品中的椰毒假单胞菌。An assay using loop-mediated isothermal amplification （ LAMP） technology was developed for rapid detection of Pseudomonas cocovenenans. The 16S ～ 23S rRNA internal transcribed spacer （ ITS） sequence of Pseudomonas cocovenenans as target sequences,used to design LAMP primers.The LAMP mixture was made in 25μL of reaction mixture,also LAMP was used both in Pseudomonas cocovenenans and in tremella of Pseudomonas cocovenenans and we compaired the method with PCR.The result showed that a rapid technique for detection of Pseudomonas cocovenenans initially established by LAMP. The sensitivity of LAMP assay was 1000 times as high as PCR,the sensitivity of the LAMP was 5.4CFU /mL,and the detection limit of artificially contaminated tremella was 76CFU /g. The whole experimental procedure （ contain DNA purified,LAMP and UV transilluminator） can be completed in 2h. LAMP technology provides a simple,rapid and sensitive method for detection Pseudomonas cocovenenans in food.

关 键 词：环介导等温扩增 快速检测 椰毒假单胞菌 

分 类 号：TS207.4[轻工技术与工程—食品科学]