中华皮肤科杂志 ›› 2004, Vol. 37 ›› Issue (8): 466-468.

• 论著 • 上一篇    下一篇

新生隐球菌28S rDNA的基因型特征与序列分析

姚志荣1, 陈仁贵1, 廖万清2, 任大明3   

  1. 1. 上海第二医科大学新华医院皮肤科 200092;
    2. 上海第二军医大学长征医院皮肤科;
    3. 复旦大学遗传学研究所遗传工程国家重点实验室
  • 收稿日期:2003-08-20 出版日期:2004-08-15 发布日期:2004-08-15

Characterization and Sequence Analysis of 28S rDNA of Cryptococcus neoformans

YAO Zhi-rong1, CHEN Ren-gui1, LIAO Wan-qing2, REN Da-ming3   

  1. Department of Dermatology, Xinhua Hospital, Shanghai Second Medical University, Shanghai 200092, China
  • Received:2003-08-20 Online:2004-08-15 Published:2004-08-15

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摘要： 目的 探讨新生隐球菌各变种、血清型与基因型的关系。方法 新生隐球菌标准株10株、新生隐球菌荚膜缺陷株CAP10以及临床分离株19株,采用变性梯度胶电泳(DGGE)结合DNA序列分析,对以上菌株的28S rDNA片段进行研究。结果 经过DGGE和DNA序列分析,所有新生隐球菌新生变种(A、D型)具有一致的基因带型和序列,格特变种(B、C型)具有不同于新生变种的独特一致的带型和序列;AD型的基因型和序列与格特变种完全相同,新生隐球菌荚膜缺陷株CAP10(D型)的基因型类同于新生变种;19株临床分离菌在DGGE上可分为2型,17株与A、D型完全相同,2株与B、C型相同。结论 DGGE结合DNA序列分析,对于探讨新生隐球菌各变种、血清型的基因型特征、关系具有重要价值;我国非艾滋病隐球菌病感染以新生变种为主,AD型在系统发育上更类同于格特变种,而不是新生变种;本研究不支持A血清型为新生变种以外的新变种即grubii变种的观点。

关键词: 隐球菌,新型, DNA,真菌, 序列分析,DNA

Abstract: Objective To investigate genetic relationships among five serotypes of two variants of Cryptococcus neoformans. Methods PCR mediated DGGE (denaturing gradient gel electrophoresis) and sequence analysis of 28S rDNA of C. neoformans were performed in ten reference strains, C. neoformans capsular-deficient strain CAP10, and nineteen clinical isolates from non-HIV patients. Results The results of DGGE and analysis of nucleotide sequences of 28S rDNA showed identical patterns and nucleotide sequences in the serotype A and D of C. neoformans var. neoformans, which were distinct from the serotye B and C of C. neoformans var. gattii. The patterns and sequences of serotype AD coincided with those of C. neoformans var. gattii. The patterns and nucleotide sequences of C. neoformans capsular-deficient strain CAP10 (serotype D) and serotype A and D were identical. Of the nineteen clinical isolates, seventeen had patterns of serotype A and D, and the others had patterns of serotype B and C. Conclusions PCR mediated DGGE integrated with sequence analysis of 28S rDNA is a valuable tool for the classification of C. neoformans. The clinical isolate of C. neoformans var. neoformans is predominant in Chinese non-HIV patients. Serotype AD is genetically close to C. neoformans var. gattii rather than C. neoformans var. neoformans. The data seem not to be in favor of previous study that serotype A, C. neoformans var. grubii, is a new variant of C. neoformans.

Key words: Cryptococcus neoformans, DNA, fungal, Sequence analysis, DNA