副溶血弧菌LAMP检测方法的建立

被引：70

作者：

徐芊

孙晓红

赵勇

潘迎捷

机构：

[1] 上海水产大学食品学院

来源：

中国生物工程杂志 | 2007年 / 12期

关键词：

副溶血弧菌; 环介导等温扩增技术(LAMP); 不耐热溶血毒素基因(tlh); 检测;

D O I：

10.13523/j.cb.20071214

中图分类号：

R155.5 [食品卫生与检验];

学科分类号：

摘要：

副溶血弧菌(Vibrioparahaemolyticus)是一种能引起食源性疾病的重要病原菌。首次将一种新颖的核酸扩增技术-环介导等温扩增技术(loop-mediatedisothermalamplification,LAMP)应用于副溶血弧菌的快速检测。针对副溶血弧菌不耐热溶血毒素基因(tlh)设计4条特异性引物(两条内引物和两条外引物)进行LAMP扩增,对扩增反应进行优化,最佳反应时间为60min,反应温度为60℃。对12种细菌共28株菌进行LAMP扩增,仅14株副溶血弧菌得到阳性扩增结果,证明引物具有很高的特异性。副溶血弧菌基因组DNA和纯培养物的检测灵敏度分别约为90fg和24cfu/ml。对模拟食品样品进行直接检测,检测限为89cfu/g。结果表明,该方法检测副溶血弧菌特异性强、灵敏度高,并且操作简便、检测成本低,1小时即可完成,有望发展成为快速检测副溶血弧菌的有效手段。

引用

页码：66 / 72

页数：7

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