摘要
应用聚合酶链式反应(Polymerase Chain Reaction,PCR)技术,建立能在12h内快速检测大肠杆菌的体系。把环境水样过滤浓缩后,将截留在膜上的细菌直接或8h增菌后提取DNA模板,以大肠杆菌Uid A基因设计引物,经过PCR扩增、凝胶电泳检测,水环境样品在不增菌条件下和增菌条件下检出限分别为1.52×104个/L,1个/L。与滤膜法比较,PCR法时间缩短、灵敏度和准确度提高,适宜于水环境中大肠杆菌的快速检测。
To rapidly detect Escherichia coli (E.coli) in water environment in 12 hours, a PCR-based system was established. The water sample was filtered by membrane, then the DNA template was extracted from the E.eoli left on the membrane directly or after 8 hours enrichment. The primers were designed from the E.coli Uid A gene, and PCR amplification, agarose gel electrophoresis was conducted. The results showed that the detection limits of E.coli in water sample with and without 8 hours enrichment were 1.52×10^4CFU/L and 1CFU/L respectively. Compared with the membrane filter method, PCR detection method has higher sensitivity and accuracy and is time saving.
出处
《环境科技》
2010年第2期52-54,58,共4页
Environmental Science and Technology
关键词
大肠杆菌
快速检测
聚合酶链式反应
Escherichia eoli
Rapid detection
Polymerase Chain reaction (PCR)
E.coli