中华皮肤科杂志 ›› 2005, Vol. 38 ›› Issue (8): 503-505.

• 论著 • 上一篇    下一篇

聚合酶链反应检测深部致病真菌的实验研究

刘军, 刘维达   

  1. 中国医学科学院、中国协和医科大学皮肤病研究所真菌科 南京 210042
  • 收稿日期:2004-10-25 出版日期:2005-08-15 发布日期:2005-08-15
  • 通讯作者: 刘维达,E-mail:liumyco@hotmail.com E-mail:liumyco@hotmail.com
  • 基金资助:
    江苏省卫生厅医学科技发展基金重大课题(H200203)

Detection of Deep Pathogenic Fungi by PCR

LIU Jun, LIU Wei-da   

  1. Institute of Dermatology, Chinese Academy of Medical Sciences & Peking Union Medical College, Nanjing 210042, China
  • Received:2004-10-25 Online:2005-08-15 Published:2005-08-15

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摘要： 目的 建立能用于临床实践的检测常见致病真菌的聚合酶链反应(PCR)方法.方法 设计了以热启动PCR为基础的实验方法,首先用真菌通用引物对标本进行单重PCR,若阳性,再用白念珠菌、烟曲霉和新生隐球菌的种特异性引物进行三重PCR来检测这3种常见致病真菌.结果 对9属55种78株常见深部真菌均扩增出260bp的DNA片段,而对细菌和人DNA均未扩增出目的 片段,具有高度特异性和敏感性,该方法操作简便且成本低.结论 以热启动PCR为基础的单重PCR和三重PCR方法可能成为临床上深部真菌感染理想的快速诊断工具.

关键词: 聚合酶链反应, 念珠菌属, 曲霉菌属, 隐球菌属

Abstract: Objective To develop a PCR method to detect common deep pathogenic fungi for the application in clinical settings.Methods A hot-start polymerase chain reaction (PCR)-based method was developed.A simplex PCR with a universal fungus-specific primer set was used to detect experimental and simulating clinical specimens.For specimens with positive results,a triplex PCR was further applied with the species-specific primers of Candida albicans,Aspergillus fumigatus and Cryptococcus neoformans.Results A 260 bp product was successfully amplified from all 78 strains of 55 fungal species in 9 fungal genera,but not amplified from any strain of other microbes and human cells.The detection system was of high specificity and sensitivity,and convenient and inexpensive for use.Conclusion The simplex and triplex PCR approach based on highly specific hot-start PCR might be useful for quick detection of deep fugal infection in routine clinical laboratory practice.

Key words: Candida, Aspergillus, Cryptococcus, Polymerase chain reaction