环介导等温扩增技术检测创伤弧菌方法的建立与应用

食品科学 ›› 2012, Vol. 33 ›› Issue (20): 262-264.

环介导等温扩增技术检测创伤弧菌方法的建立与应用

薛超波，许镇坚，黄朱梁，孙瑛

舟山市质量技术监督检测研究院

收稿日期:2011-08-11 修回日期:2012-09-20 出版日期:2012-10-25 发布日期:2012-11-09
通讯作者: 薛超波 E-mail:xuechaobo2001@163.com
基金资助:
浙江省分析测试科技计划项目;浙江省科技厅项目

Development and Application of Loop-Mediated Isothermal Amplification (LAMP) Method for Detection of Vibrio parahaemolyticus

Received:2011-08-11 Revised:2012-09-20 Online:2012-10-25 Published:2012-11-09
Contact: Chao-bo XUE E-mail:xuechaobo2001@163.com
Supported by:
;Grant No.2010C32005

摘要/Abstract

摘要： 根据创伤弧菌vvhA基因设计4条特异性引物，采用环介导等温扩增方法(LAMP)建立创伤弧菌快速检测方法。结果表明，建立的LAMP方法具有高度特异性，经对13株细菌进行扩增，创伤弧菌为LAMP阳性，其他菌株均为LAMP阴性。该方法对培养的创伤弧菌的检测下限为51CFU/mL，可在60min内完成扩增反应。用建立的LAMP方法对120份海产品进行检测，共检出37份LAMP阳性样本，与传统培养方法结果一致。该方法与传统方法相比，可节省大量时间，且对实验仪器和操作人员的要求低，具有良好的实用性。

关键词: 创伤弧菌, 环介导等温扩增技术, vvhA基因

Abstract: In order to establish a loop-mediated isothermal amplification (LAMP) method for rapid diagnosis of Vibrio vulnificus, 4 primers that can recognize the vvhA gene of Vibrio vulnificus were designed and used for LAMP assay. Results showed that this method was highly specific for Vibrio vulnificus. Among 13 amplified strains, Vibrio vulnificus was LAMP positive, and other strains were LAMP negative. Moreover, the detection procedure could be completed within 60 minutes and had a detection limit of 51 CFU/mL. Of 120 seafood samples detected by this method, 37 were positive, which was consistent with the result obtained by the NMKL method (No. 156. 2nd edition. 1997). Compared with the NMKL method, this LAMP method was greatly time saving and lowered instrumental and operational requirements, suggesting good practical applicability.

Key words: Vibrio vulnificus, loop-mediated isothermal amplification (LAMP), vvhA gene

中图分类号:

TS201.3

薛超波，许镇坚，黄朱梁，孙?瑛. 环介导等温扩增技术检测创伤弧菌方法的建立与应用[J]. 食品科学, 2012, 33(20): 262-264.

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