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中国真菌学杂志 2014, Vol. 9  Issue (5): 264-267.

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不同毒力差异基因型的新生隐球菌MLST分型及交配型鉴定研究

康颖倩, 陈玉如, 王梅竹, 赵亮, 李小玲, 金方, 刘涛华   

  1. 贵阳医学院微生物教研室, 贵阳 550004
  • 收稿日期:2014-09-22 出版日期:2014-10-28 发布日期:2014-10-28
  • 作者简介:康颖倩, 女 (汉族), 博士, 副教授.E-mail:joycekangtokyo@qq.com
  • 基金资助:
    国家自然基金地方项目(31060006,31260029);贵州省科技厅国际合作项目(黔科合外G字[2009]700114号);贵州省社会发展科技攻关项目(黔科合SY字[2011]3017号);贵阳市科技局社会发展与民生计划(筑科合同[2011103]16号)

MLST genotyping and mating type identification of C.neoformans in genotypes with virulent difference

KANG Ying-qian, CHEN Yu-ru, WANG Mei-zhu, ZHAO Liang, LI Xiao-ling, JIN Fang, LIU Tao-hua   

  1. Department of Microbiology, Guiyang Medical University, Guiyang 550004
  • Received:2014-09-22 Online:2014-10-28 Published:2014-10-28

摘要: 目的 了解及比较两组毒力差异明显的新生隐球菌格鲁比变种的多位点序列分型 (MLST)的特点并进行交配型鉴定.方法 采用多位点序列分型 (MLST)的方法,设计7个看家基因 (CAP59,GPD1,LAC1,PLB1,SOD1,URA5IGS1)的引物, 扩增并分析来源分别为环境和临床的各10株新生隐球菌格鲁比变种的基因型,并鉴定实验菌株交配型,与多位点微卫星分型 (MLMT)结果对比,比较不同基因分型方法在分类中的稳定性和可靠性.结果 在微卫星分型中为MLMT-36的10株环境分离株,MLST分型为ST-15,而在微卫星分型中为MLMT-13型的10株临床分离株,MLST分型为ST-32,所有菌株交配型均为MAT-α.结论 MLST分型结果与MLMT分型结果高度一致,提示以上两种分子分型技术在真菌分类鉴定研究中可显示对于其分离背景及进化来源的高分辨率及稳定性.

关键词: 新生隐球菌, MLST分型, MLMT分型, 交配型鉴定

Abstract: Objectives To study the Multilocus Sequence Typing (MLST) characteristics of C.neoformans var.grubii and to identify their mating types.Methods Primers of 7 housekeeping genes (CAP59,GPD1,LAC1,PLB1,SOD1,URA5 and IGS1) were designed for PCR amplification; the genotypes of 20 environment and clinical isolates were analyzed with MLST result and the matting types of these strains were identified. And both the results of MLST and Multilocus Microsatellite Typing (MLMT) were compared to each other for evaluating their stability and reliability in taxonomy.Results The multilocus microsatellite genotype of all the 10 environment isolates was MLMT-13 in previous experiment and the MLST type of there strains was ST-15 in this experiment. Similarly, all the MLMT-36 clinical isolates belong to ST-32. And all the strains were MAT-α.Conclusion The MLST Results are highly consistent with MLMT Results in previous experiment which reveals that both molecular typing Methods are high resoluted and stable on classification of different isolation background on species or varieties level.

Key words: Cryptococcus neoformance, MLST, MLMT, mating type

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