多酶恒温核酸快速扩增技术在霍乱弧菌检测中的应用  

Application of Multi-enzyme Isothermal Rapid Amplification in the Detection of Vibrio Cholera

作　　者：李盛杰 江海涛 LI Sheng-jie;JIANG Hai-tao(School of Food Science,Nanjing Xiaozhuang University,Nanjing 211171,China)

机构地区：[1]南京晓庄学院食品科学学院,江苏南京211171

出　　处：《南京晓庄学院学报》2021年第6期49-52,共4页Journal of Nanjing Xiaozhuang University

基　　金：江苏省高校重点实验室项目(苏教科〔2016〕8).

摘　　要：霍乱弧菌(Vibriocholera)是水产养殖中常见的致病菌,建立起快速、灵敏、便捷的检测方法对加强水产养殖病害监测和防控工作有重要意义.本研究利用多酶恒温核酸快速扩增技术,根据霍乱弧菌外膜蛋白基因ompW的保守序列,设计6对引物组,对引物特异性进行验证;并设计不同的模板浓度梯度,以验证该方法的灵敏度.结果表明,本研究确定的引物对霍乱弧菌检测的特异性强,灵敏度高(可低至10 fg/μL),无需特殊设备,适合实验室或者现场的快速检测要求.Vibrio cholera is a common pathogen in aquaculture.The establishment of fast,sensitive and convenient detection methods is of great significance to strengthen the monitoring and prevention of diseases in aquaculture.In this study,we designed 6 pairs of primer sets based on the conservative sequence of the Vibrio cholera virulence gene ompW to verify the specificity of these primers in multi-enzyme isothermal rapid amplification;and designed different template concentration gradients to verify the method’s sensitivity.The results showed that the primers in this study have strong specificity and high sensitivity(as low as 10 fg/μL)for the detection of Vibrio cholera,without special equipment.And this method is suitable for rapid detection requirements in the laboratory or on-site.

关 键 词：霍乱弧菌 多酶恒温核酸快速扩增技术 外膜蛋白基因 

分 类 号：Q503[生物学—生物化学]