PMA与PCR结合的细菌活细胞检测方法

被引:26
作者
罗剑飞
林炜铁
郭勇
机构
[1] 华南理工大学生物科学与工程学院
关键词
叠氮溴化丙锭; 脱氧核糖核酸; 聚合酶链反应; 活细胞检测;
D O I
暂无
中图分类号
Q93-3 [微生物研究与微生物实验];
学科分类号
071005 ; 100705 ;
摘要
基于PCR的分子生物技术在检测过程中不能有效区分样品中细菌的死活状态,这会导致对细菌数量的错误估计.文中用叠氮溴化丙锭(PMA)对样品基因组提取进行前处理,使PMA与样品中死细胞的DNA分子共价交联,并抑制该DNA分子的PCR扩增.结果表明:当样品中PMA质量浓度大于3μg/mL、曝光时间大于3 min时,PMA可抑制细胞膜破裂的E.coli死细胞DNA的PCR扩增;PMA质量浓度高于50μg/mL时对活细胞DNA的PCR扩增有一定影响;当浊度小于10NTU时,PMA能有效地抑制死细胞DNA的PCR扩增,而当浊度大于100NTU时,PMA失去效果.
引用
收藏
页码:142 / 146
页数:5
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