作　　者： ; ; ; ; ; ;

机构地区： 华南理工大学轻工与食品学院

出　　处： 《食品科学》 2012年第22期217-220,共4页

摘　　要： 建立将叠氮溴化丙锭(PMA)与定量PCR(qPCR)技术结合,用于检测热灭活菌背景条件下的大肠杆菌活菌的方法。结果表明:5min以上的强烈光照可以使PMA与DNA共价交联,同时完全钝化游离的PMA以避免"假阴性"结果;抑制死菌DNA扩增的最低PMA质量浓度为10μg/mL,不抑制活菌DNA扩增的最高PMA质量浓度为20μg/mL。当活菌/总菌大于1%时,经PMA预处理,可以消除热灭活菌DNA的干扰,实现对活菌的定量检测。 In this work, a method to detect live E. coliO157：H7wasestablishedusingpropidiummonoazide（PMA）coupled with quantitative polymerase chain reaction （qPCR）. The minimum inhibitory concentration against DNA amplification from dead bacteria was 10 ~tg/mL, and PMA did not inhibit DNA application from live bacteria at concentrations equal to or lower than 20 ~tg/mL. Covalent cross-linking of PMA with DNA was induced by strong light illumination for 5 min and meanwhile, free PMA was completely inactivated, resulting in the avoidance of false negative results. When the live to dead bacteria ratio was more than 1%, PMA pretreatment allowed the elimination of DNA interference from thermally inactivated bacteria and therefore live bacteria could be quantified.

关 键 词： 叠氮溴化丙锭 定量 活菌 检测

领　　域： [轻工技术与工程] [轻工技术与工程]