基因释放剂法快速提取病原性真菌DNA直接用于PGR扩增的研究

Rapid preparation of DNA from pathogenic fungi by using gene releaser for direct polymerase chain reaction

目的：利用基因释放剂快速制备病原性真菌DNA模板,简化PCR方法和减少标本用量。方法：利用基因释放剂提取临床样品中酵母菌DNA,直接用于PCR扩增,并与常用的氯化苄提取法进行比较。结果：该方法成功用于9种临床常见致病性酵母菌PCR扩增,敏感性达10个孢子/mL。用于含菌血标本的检测,均获得满意的结果。结论：该技术检测时间短、操作简单、成本低等优点,非常适合用于临床标本中致病性酵母菌DNA模板的快速制备。

Objective：In order to simplify PCR and reduce the amount of samples,gene releaser was used to prepare PCR template of pathogenic fungi without extraction of DNA.Methods： DNA of pathogenic yeast were extracted with gene releaser and used directly for polymerase chain reaction.This extraction method was compared with the method with benzyl chloride.Results： This method was successful for PCR amplification of nine kinds of pathogenic yeast.The sensibility of the PCR was about 10 cells/ml.Satisfactory results had been obtained in detection of pathogenic yeast in blood.Conclusion： This method is a simple,fast,sensitive and low cost technology.Gene releaser is a very useful agent for the rapid and simplified direct PCR amplification without extraction of DNA.