摘要
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.
出处
《激光杂志》
CAS
CSCD
北大核心
2010年第4期78-79,共2页
Laser Journal
基金
重庆市自然基金(项目编号:CSTC,2007BB5080)
军队医药卫生科研基金课题(项目编号:06G073)
关键词
基因释放剂
聚合酶链反应
酵母菌
真菌
gene releaser
polymerase chain reaction
yeast
medical fungi