中华皮肤科杂志 ›› 1998, Vol. 31 ›› Issue (5): 295-297.

• 论著 • 上一篇    下一篇

新生隐球菌随机扩增DNA指纹分型研究

温海1, 廖万清1, 姚志荣1, 陈江汉1, 吴建华1, M. A. Viviani2   

  1. 1. 第二军医大学附属长征医院皮肤科真菌病研究室, 上海200003;
    2. 意大利米兰大学卫生和预防医学研究所
  • 收稿日期:1997-04-17 修回日期:1997-08-27 出版日期:1998-10-15 发布日期:1998-10-15
  • 基金资助:
    意大利米兰大学卫生和预防医学研究所专项资助

Typing of Cryptococcus neoformans by Random Amplification of DNA Fingerprinting

Wen Hai1, Liao Wanqing1, Yao Zhirong1   

  1. Cryptococcus Laboratory, Department of Dermatology, Changzheng Hospital attached to the Second Military Medical University, Shanghai 200003
  • Received:1997-04-17 Revised:1997-08-27 Online:1998-10-15 Published:1998-10-15

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摘要： 目的 用随机引物扩增新生隐球菌临床和环境分离株的DNA,观察DNA指纹与血清型的关系,探索新生隐球菌基因分型标准。方法 采用3种不同寡核苷酸重复序列[CN1(GTG)5、CN2(GACA)4和CN3(GATA)4]为引物,对从临床和鸽粪中分离到的24株血清A型和8株四种血清型的新生隐球菌标准株的DNA进行扩增,根据扩增产物的电泳带型来对比DNA指纹和血清型之间的关系。结果 24株临床和鸽粪分离的血清A型株中,20株产生和标准株完全相似的DNA带型,1株与标准D型DNA带型相似。3株的PCR指纹与血清A、B、C、D型均不相同。2株B型和2株C型产生几乎完全一致的DNA指纹带型。结论 ①大多数血清A型新生隐球菌菌株具有相似DNA指纹带型。②血清B型和C型(gattii变种)的DNA带型不易区别。③相同血清型不同的株可能有不同的DNA指纹带型。④PCR法用于新生隐球菌菌株鉴定是一种快速、方便、可行的方法。

关键词: 新生隐球菌, PCR, 随机引物

Abstract: Objective To investigate the DNA typing, observe relationship between DNA fingerprinting patterns and serotypes of Cryptococcus neoformans, and find a suitable genotyping standard for Cryptococcus neoformans. Methods Three primers, including CN1(GTG)5, CN2(GACA)4 and CN3(GATA)4, were used to distinguish variations among strains of C.neoformans. Results The distinguishable fingerprinting bands for serotype of C.neoformans were yielded by primer CN2. Using this primer, of 24 clinical and environmental isolates of serotype A of C.neoformans and 8 standard strains investigated by PCR, 20 strains produced complete identical fingerprinting patterns, the other 4 strains had different fingerprinting patterns. 2 strains of serotype B and C yielded indistinguishable fingerprinting patterns. Conclusion ①The majority of strains of serotype A had similar and stable fingerprinting patterns. ②Some strains of serotype B and C had an indistinguishable fingerprinting patterns. ③The same serotype strains from different sources may produce different DNA fingerprinting patterns. ④The DNA fingerprinting is a rapid, simple and feasible method for identifying Cryptococcus neoformans.

Key words: Cryptococcus neoformans, PCR, Primer