利用环介导等温扩增技术快速检测饮用水中的大肠杆菌

Rapid Detection of Escherichia coli in Purified Water by Loop-Mediated Isothermal Amplification Technology

本研究以饮用水为研究对象,利用大肠杆菌mal B保守基因设计LAMP特异引物,通过优化反应条件,建立了适用于快速检测饮用水中大肠杆菌的LAMP方法,可在60℃条件下反应60 min完成检测,检测灵敏度达到1.35 CFU/m L。利用SYBR GreenⅠ染料剂可在紫外灯下直接观察检测结果。通过检测掺入已知大肠杆菌的纯净水样品,结果表明,本研究所建立的LAMP方法具有特异性强、灵敏度高、操作简便的特点,可用于饮用水中大肠杆菌致病菌的快速检测,对预防和控制大肠杆菌的传染具有重要意义。

In this study, with the specific primers designed according to the conserved region of malB gene of Escherichia coli, a rapid method of loop - mediated isothermal amplification for detecting Escherichia coli in purified water was established through optimizing the reaction conditions. For one time detection, it could be completed at 60℃ in 60 minutes, and the detection sensitivity was 1.35 CFU/mL. The detection results could be observed under UV lamp with SYBR Green I . By detecting the purified water samples incorporated with known Escherichia coli, the results showed that the established LAMP method owned advantages of strong specificity, high sensitivity and easy to operation. It was suitable for rapid detection of pathogenic Escherichia coli in purified water, and it had important significance for prevention and control of infection from Escherichia coll.