国内首例播散性毛孢子菌病病原菌阿萨希丝孢酵母菌的DNA序列分析  

26S rDNA D1/D2 domain sequence analysis of the pathogenic strain from the first case of disseminated trichosporonosis in China

作　　者：杨蓉娅[1] 李大伟[1] 赵建花[2] 敖俊红[1] 王文岭[1] 白逢彦[2] 

机构地区：[1]解放军北京军区总医院皮肤科,100700 [2]中国科学院微生物研究所

出　　处：《中华医学杂志》2001年第8期472-475,共4页National Medical Journal of China

摘　　要：目的　鉴定播散性毛孢子菌病病原丝孢酵母菌AS 2 .2 174,并探讨将DNA序列分析用于病原酵母菌快速鉴定的可行性。方法　丝孢酵母菌株取自 1例播散性毛孢子菌病患者的皮肤病损处。用简易微量DNA提取方法从病原酵母菌株中提取细胞总DNA ,用PCR方法扩增 2 6SrDNAD1/D2区域 ,纯化后用热循环测序法对扩增产物进行直接测序。结果　测定了菌株AS 2 .2 174的 2 6SrDNAD1/D2区域的 6 0 4个碱基序列 ,并将该序列登录于GenBank (登录号 :AF337949)。通过与丝孢酵母属所有种模式菌株D1/D2序列的比较分析表明 ,AS 2 .2 174与阿萨希丝孢酵母 (Trichosporonasahii)的序列完全相同 ,而与在表型特征上与Trichosporonasahii难以区分的另外 3个丝孢酵母菌种或变种分别相差 3～ 8个碱基。结论　确定菌株AS 2 .2 174属于Trichosporonasahii,并显示DNA序列分析在病原酵母菌鉴定方面具有常规方法难以达到的快速和准确。Objective To determine the large subunit (26S) rDNA D1/D2 domain sequence of AS 2 217 4, the pathogenic strain from the first case of disseminated trichosporosis in China, and study the feasibility of applying DNA sequencing in rapid identification of pathogenic yeast. Method The strain of Trichosporon (AS 2 217 4) was collected from the skin lesion of a case of disseminated trichosporosis Nuclear DNA was extracted from the cells of AS 2.2174 by a simplified protocol and the 26S rDNA D1/D2 domain was amplified by PCR. The PCR product was then directly sequenced with the ABI PRISM BigDye cycle sequencing kit. Results The 26S rDNA D1/D2 domain of strain AS 2.2174 was determined (GenBank accession number: AF337949). Comparison analysis indicated that the D1/D2 sequence of AS 2.2174 is identical with that of the type strain of Trichosporon asahii , and differs from those T. asteroides, T. faecale and T. coremiiforme in 3, 5 and 8 nucleotides, respectively. The latter three species are phenotypically indistinguishable from T. asashii . Conclusion The strain AS 2.2174 was confirmed to belong to Trichosporon asahii . This species is the first record in China clinically and taxonomically. DNA sequencing shows an advantage in fast and accurate identification of clinically relevant yeast species.

关 键 词：毛孢子菌病 酵母菌 序列分析 DNA 

分 类 号：R379[医药卫生—病原生物学]