中华皮肤科杂志 ›› 2005, Vol. 38 ›› Issue (8): 495-497.

• 论著 • 上一篇    下一篇

利用核糖体基因进行申克孢子丝菌基因分型

张振颖1, 刘晓明1, 高晓蓉2, 杨国玲1, 金礼吉2, 安利佳2   

  1. 1. 大连医科大学附属第一医院皮肤科教研室 116011;
    2. 大连理工大学环境生命学院
  • 收稿日期:2004-09-07 发布日期:2005-08-15
  • 通讯作者: 刘晓明,E-mail:LXM_1956@Yahoo.com.cn E-mail:LXM_1956@Yahoo.com.cn
  • 基金资助:
    国家自然科学基金资助项目(30470104)

DNA Typing of Sporothrix schenckii by Analysis of Ribosomal-DNA Regions

ZHANG Zhen-ying1, LIU Xiao-ming1, GAO Xiao-rong2, YANG Guo-ling1, JIN Li-ji2, AN Li-jia2   

  1. Department of Dermatology, First Afilliated Hospital, Dalian Medical University, Dalian 116011, China
  • Received:2004-09-07 Published:2005-08-15

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摘要： 目的 利用探针与DNA印迹法对申克孢子丝菌进行种内分型,探讨其基因型特征与菌种来源及临床表现的关系.方法 CTAB法提取来源于不同地区31株孢子丝菌临床分离株及1株标准株的基因组DNA,以真菌通用引物ITS4、NS5扩增标准株的rDNA序列作为探针,与经限制性内切酶ApaI酶切后的基因组DNA进行印迹杂交.结果 杂交后形成多种清晰而稳定的带型,根据带型将31株孢子丝菌分为15种基因型(A-O型),其中A、B、C三型占51.61%.结论 探针与DNA印迹法是孢子丝菌种内分型较为敏感而可靠的方法,该方法所分基因型的不同与菌种的地区来源及临床表现有一定的相关性.

关键词: 孢子丝菌属, DNA,核糖体, 基因型, 印迹法,DNA

Abstract: Objectives To identify genotypes of 31 Sporothrix schenckii (S.schenckii) strains by Southern blotting and to explore the relationship between genotypes and geographic distributions and clinical manifestations.Methods Total DNA was extracted by cetyltriethyl ammonium bromide (CTAB).The polymorphisms were detected by hybridization of ApaⅠ-digested S.schenckii genomic DNA with a probe amplified from the small-subunit rDNA and adjacent internal transcribed spacer (ITS) regions.The band patterns manifested by Southern blotting were employed to investigate genotypes of 31 strains of S.schenckii collected from five different areas in China.Results Of 31 strains of S.schenckii,15 individual patterns (DNA Type A-O) were recognized.Type A to C accounted for 51.61% of all strains.Conclusion The Southern blotting provides a highly sensitive and reliable means for DNA typing of S.schenckii.It is also found that there is an obvious correlation between DNA patterns and different geographic distribution and clinical manifestations.

Key words: Sporothrix, DNA,ribosomal, Genotype, Blotting,Southern