rDNA—RFLP多酶切技术用于人类致病相关毛孢子菌种问鉴定初探  

Primary exploration on identification of pathogenic Trichosporon spp. with rDNA-RFLP analysis

作　　者：吕雪莲[1] 张晓利[2] 王淼淼[1] 刘泽虎[1] 沈永年[1] 刘维达[1] 

机构地区：[1]大连市皮肤病医院,116021 [2]中国医学科学院北京协和医学院皮肤病研究所

出　　处：《中华皮肤科杂志》2010年第8期534-537,共4页Chinese Journal of Dermatology

基　　金：科技部重大专项（2008ZX10004-002）;2007-2009年度卫生部部属（管）医院临床学科重点项目（第7号）;卫生部公益性行业科研专项经费（200802032）

摘　　要：目的 探讨rDNA-ITS/IGS1-RFLP多酶切技术构建毛孢子菌的酶切谱系并进行聚类分析,尝试将该方法用于人类致病相关毛孢子菌的快速种间鉴定.方法 收集8种14株毛孢子菌,对其进行rDNA-ITS/IGS1扩增测序,并分别应用HaeⅢ、Hha Ⅰ、HaeⅢ和Hha Ⅰ双酶、HinfⅠ、Msp Ⅰ和Taq Ⅰ对扩增片段进行RFLP酶切.结果 rDNA-ITS-RFLP多酶切可将8种毛孢子菌分为不同进化分支的4个亚类,而rDNA-IGS1-RFLP多酶切则可实现8种14株毛孢子菌准确的种间鉴定,并实现对部分进化距离较远的阿萨希毛孢子菌基因型之间的区分.结论 rDNA-ITS/IGS1-RFLP多酶切技术有望用于毛孢子菌属种水平的快速鉴定.Objective To genotype Trichosporon spp. with rDNA-ITSAGSl-RFLP analysis followed by cluster analysis, and attempt to apply this method to rapid species identification of human pathogenic Trichosporon spp.. Methods Fourteen strains of Trichosporon, which belonged to 8 species, were collected. The rDNA-ITS/IGSl regions were amplified by PCR and sequenced. Simultaneously, the amplicons were digested separately with restriction enzymes, including Hae III, Hha I , Hae IH and Hha I , Hinf I , Msp I and Taq I . Results The 8 species of Trichosporon could be classified into 4 subgroups with rDNA-ITS-RFLP, while inter-species identification of all the 14 strains from 8 species of Trichosporon could be realized with rDNA-IGSl-RFLP. Also, those genotypes of T. asahii which had relative long phylogenic distance could even be discriminated with rDNA-IGSl-RFLP. Conclusion The rDNA-ITS/IGSl-RFLP analysis is expected to be used in rapid interspecific identification of genus Trichosporon.

关 键 词：毛孢子菌属 DNA 核糖体 多态性 限制性片段长度 

分 类 号：R379[医药卫生—病原生物学]