中华皮肤科杂志 ›› 2005, Vol. 38 ›› Issue (8): 492-494.

• 论著 • 上一篇    下一篇

马拉色菌临床株分类鉴定的研究

崔凡1, 陶诗沁2, 沈永年1, 吕桂霞1, 陈伟1, 李筱芳1, 胡素泉1, 杨丽佳2, 刘维达1   

  1. 1. 中国医学科学院、中国协和医科大学皮肤病研究所 南京 210042;
    2. 无锡市第二人民医院皮肤科
  • 收稿日期:2004-09-24 发布日期:2005-08-15
  • 通讯作者: 刘维达,E-mail:liumyco@hotmail.com E-mail:liumyco@hotmail.com

Genotyping of Clinical Isolates of Malassezia from Patients with Tinea Versicolor by PCR-RFLP

CUI Fan1, TAO Shi-qin2, SHEN Yong-nian1, LÜ Gui-xia1, CHEN Wei1, LI Xiao-fang1, HU Su-quan1, YANG Li-jia2, LIU Wei-da1   

  1. Institute of Dermatology, Chinese Academy of Medical Sciences & Peking Union Medical College, Nanjing 210042, China
  • Received:2004-09-24 Published:2005-08-15

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摘要： 目的 用PCR-RFLP方法对马拉色菌临床株快速分类,并结合生化鉴定分析结果的可靠性.方法 先用吐温试验和过氧化氢酶试验对74株分离自花斑癣的马拉色菌临床株和7株标准菌株进行分种,再用特异性引物扩增马拉色菌的28SrDNA,对扩增产物分别用Eco88Ⅰ、Bsp143Ⅱ和BshNⅠ进行RFLP分析.结果 3种限制性内切酶成功地将限制、钝形和厚皮马拉色菌鉴定出来,并发现限制马拉色菌在基因同源性上与其他6种马拉色菌存在较大差异.此外通过RFLP法还纠正了生化分型的错误.结论 PCR-RFLP法是一种有希望能够快速、准确对马拉色菌属进行种间分类的分子生物学技术.

关键词: 马拉色霉菌属, 花斑癣

Abstract: Objectives To develop a rapid genotyping method of clinical isolates of Malassezia from patients with tinea versicolor by PCR-RFLP,and to evaluate reliability of the approach as compared with biochemical classification.Methods Tween assimilation test and catalase reaction were carried out to identify 74 isolates of Malassezia species from patients with tinea versicolor and 7 Malassezia reference strains.The sequence of 28S rDNA of Malassezia species was amplified by PCR,and then the product was analyzed by RFLP with Eco88I,Bsp143Ⅱ and BshNⅠ,respectively.Results M.restricta,M.obtusa and M.pachydermatis were successfully identified by three restriction endonucleases.M.restricta was found to be more diverse from the other 6 species in genetic homology.By comparison with PCR-RFLP technique,a possible mistake was discovered with biochemical method.Conclusion PCR-RFLP is a promising molecular biological technique,which could rapidly and correctly classify Malassezia species.

Key words: Malassezia, Tinea versicolor