食品中拟态弧菌变性高效液相色谱检测技术研究

应用PCR 结合变性高效液相色谱(DHPLC)技术对食品中致病菌--拟态弧菌进行检测，探讨其灵敏度特异性，建立快速高通量检测食品中拟态弧菌的新方法。利用DHPLC 在非变性温度下进行双链DNA 分离的原理，应用DHPLC 技术分析拟态弧菌特异性PCR 扩增产物。在优化的分析条件下，对样品洗脱峰排列形成的聚类分析图进行分析，得到拟态弧菌的特征峰型图谱。结果表明该方法有很好的特异性。与传统检测方法进行比较该方法准确度为100%。本实验建立的食品中拟态弧菌PCR-DHPLC 检测技术，特异性灵敏，准确度高，操作快速、简便，在食品安全检测中具有重要应用价值。

Study on Determination of Vibrio mimicus in Food by Polymerase Chain Reaction Coupled with Denaturing High Performance Liquid Chromatography (PCR-DHPLC)

PCR-DHPLC was adopted as a sensitive, specific, rapid and high flux approach for the detection of food-borne pathogenic bacteria, Vibrio mimicus in this study. DHPLC was used for separating double-stranded DNA under non-degeneration temperature and for detecting the specific PCR products. Under the optimum conditions, the clustering analysis diagram of elution peaks of samples were analyzed, and the spectrum with characteristic peak of Vibrio mimicus was obtained. The results showed that PCR-DHPLC has excellent specificity and accuracy of 100% compared with the traditional technology, therefore it has extensive application value in food safety assessment.