基因释放剂在快速制备医学真菌DNA中的应用

Application of the Gene Releaser in Rapid Extraction of the DNAs from Medical Fungi

目的 评价基因释放剂在快速制备医学真菌DNA中的作用和应用条件。方法 样品加入基因释放剂后分别用程序加热法、微波处理法和煮沸法处理 ,然后用 18srDNA通用引物进行PCR扩增。优化PCR扩增体系和条件。结果 3种方法均能达到实验要求 ,其中以微波处理法中的 5档火力 7min条件效果最好 ,各菌株均可被扩增出一条 482bp的DNA带。敏感性为 10 1个孢子 /ml。结论 使用基因释放剂可简便快速地从小量致病真菌样本中制备DNA 。

Objective To evaluate the use and application conditions of the gene releaser in rapid extraction of the DNAs from the specimens of medical fungi.Method After the gene releaser added, the specimens were disposed by using the procedural heating, microwave disposing and boiled methods, respectively. PCRs were carried out with a pair of primer from fungal 18s rRNA, and the system and conditions of the amplification were optimized.Results These 3 methods alll achieved the goal of the PCR test, and condition of microwave, 5 grade power and 7 minutes, was the best. A 482 bp DNA band could be amplified in all strains tested. The sensibility of the PCR was about 10 1 cells/ml. Conclusion Gene releaser can extract DNA from a little ammont of fungal specimens rapidly and simply, and can be used for the direct PCR diagnosis for deep fungal infections.