中华皮肤科杂志 ›› 2002, Vol. 35 ›› Issue (5): 352-354.

• 论著 • 上一篇    下一篇

红色毛癣菌的基因分型研究

李乔1, 杨国玲1, 刘维达2, 李朝军3   

  1. 1. 大连医科大学第一附属医院皮肤科, 116011;
    2. 中国医学科学院、中国协和医科大学皮肤病研究所;
    3. 南京师范大学
  • 收稿日期:2001-01-31 出版日期:2002-10-15 发布日期:2002-10-15

DNA Typing of Trichophyton Rubrum

LI Qiao1, LIU Weida1, YANG Guoling2, LI Zhaojun3   

  • Received:2001-01-31 Online:2002-10-15 Published:2002-10-15

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摘要： 目的 探讨探针与DNA印迹杂交法对红色毛癣菌的基因分型并分析其基因型与来源地区的相关性。方法 用溴化十六烷基三甲铵(CTAB)法提取DNA,以皮肤癣菌的特异性引物NS5[5'-AACTTAAAGGAATTGACGGAAG-3']与ITS4[5'-TCCTCCGCTTATTGATATGC-3']为引物,以红色毛癣菌的标准菌株为模板,用PCR法扩增出其rDNA部分18S区、ITSⅠ区、5.8S区和ITSⅡ区为探针,用随机引物法将探针标记32P,用EcoRⅠ酶切基因组DNA,采用DNA印迹的标准流程将酶切的基因组DNA与探针杂交;显示的不同带型,以此作为红色毛癣菌基因分型的依据。结果 所试49株红色毛癣菌(南京21株、大连26株、北京2株)分为20型(A-T型),其中A-C型占48.98%.南京株绝大多数为3条带,大连株大部分为4条带。结论 用ITS区域探针与DNA印迹杂交法对红色毛癣菌基因组分型敏感性强、分辨力高;南京与大连两地区红色毛癣菌DNA分型具有明显差异。DNA分型对红色毛癣菌病的流行病学、临床疗效的判定以及指导用药均具有重要价值。

关键词: 发癣菌属, 基因,真菌, 真菌学分型技术

Abstract: Objective To explore the DNA typing of Trichophyton rubrum(T.rubrum)by Southern blotting,analyze the relationship between genotype and different geographical areas.Methods Forty nine strains of T.rubrum(21 from Nanjing,26 from Dalian and 2 from Beijing)were studied.DNA was extracted using cetyltrimethyl ammonium bromide(CTAB).The polymorphisms were detected by hybridization of EcoRⅠ-digested T.rubrum genomic DNA with a probe amplified from the small-subunit rDNA and adjacent internal transcribed spacer(ITS)regions.The probe was made using a pair of primers NS55'-AACTTAAAGGAATTGACGGAAG-3'3 and ITS45'-TCCTCCGCTTATTGATATGC-3'3 and was labeled by 32P.The band patterns obtained by Southern blotting were used as the basis of the genotyping of T.rubrum.Results Twenty individual patterns(DNA type A to T) were recognized among 49 strains of T.rubrum.Type A to C accounted for 48.98% of all strains.The DNA patterns of Nanjing strains were represented by 3 bands,those of Dalian strains were represented by 4bands.Conclusion The DNA typing of T.rubrum using Southern blotting was sensitive,highly distinguishable.The DNA patterns of Nanjing strains are obviously diffe rent from those of Dalian strains.

Key words: Trichophyton, Genes,fungal, Mycological typing techniques