军团菌质粒快速检测方法的探讨

A Preliminary Study on A Rapid Procedure for Detection and Isolation of Plasmids in Legionella

本文通过对国内外目前应用于细菌质粒快速检测方法进行比较、筛选,结果表明:在所采用的7种方法中,4种可以检出军团菌质粒,它们是Kado和Liu法、Takahashi和Nagano法、Birnborm和Doly法和Crosa的大分子质粒提取法。鉴于这4种方法各有不足之处,参照这些方法,我们建立了一快速、简便、经济、有效的军团菌质粒检测方法。该法的关键步骤是:硷性裂解、盐析沉淀去染色体,氯仿抽提纯化、乙醇沉淀浓缩质粒DNA。此法能检出>80Md的军团菌大质粒,所得结果可直接用于酶切试验。

A comparative study was undertaken of 7 rapid procedures for the plasmid detection of Legionella. Among them, 4 were found effective. They are Kado &Liu's, Birhboim & Doly's, Takahashi & Nagano's, and Crosa's procedure for large plasmid detection of gram negative bacteria. Each of them had it's shortcoming.So a simple, rapid and effective procedure for plasmido DNA isolation was developed for Legionella without phenol. By applying this method, plasmid DNA of legionella could be lisolated and identified easily.