聚合酶链反应检测结核分支杆菌临床标本制备方法的研究

A Research on Sample Preparation for Detection of Mycobacterium Tuberculosis by Polymerase Chain Reaction

分别检测６种结核菌ＤＮＡ提取方法的裂解效能、聚合酶链反应（ＰＣＲ）扩增灵敏度及其对临床标本的阳性检出率。结果显示，ＴＥ－ＴｒｉｔｏｎＸ法和Ｃｈｅｌｅｘ－１００法能裂解绝大多数的细菌并具最高的敏感性，可检测出少至１０个菌细胞，其临床标本的阳性检出率分别为９３．３％和８６．６％；而其余４种方法效果较差，ＴＥ－Ｔｒｉｔｏｎ和Ｃｈｅｌｅｘ－１００两种沸水浴法简捷、经济、高效，值得临床实验室推广使用。

The comparision of six procedures of DNA extraction including the examination of their cell rupture efficiencies, the test of their detection limits of the PCR assay and the analysis of their positive rates in clinical samples was performed. Two boiling methods of TE-Triton and Chelex-100 showed the satisfied cell lysis efficiencies and permitted the most sensitive PCR amplification (10 cells detectable in both). When they were applied to treating the clinical samples, the highest positive rates(93.3% and 86.6%,respectively) were obtained. The four other methods were not so effictive. TE-Triton or Chelex-100 boiling method is simple, with lower cost and high efficiency and worth utilizing in clinical diagnosis laboratories.