新生隐球菌的AFLP分析

中国真菌学杂志　 2010, Vol. 5 　Issue (2): 101-104.

新生隐球菌的AFLP分析

邹先彪¹, 温海², 廖万清², 吴建华³, 仇芸², 杨金水⁴

1. 北京市解放军总医院304临床部皮肤科, 北京, 100048;
2. 第二军医大学长征医院皮肤科, 上海, 200003;
3. 第二军医大学长海医院皮肤科, 上海, 200433;
4. 复旦大学遗传所, 上海, 200433

收稿日期:2010-02-11 出版日期:2010-04-28 发布日期:2010-04-28
作者简介:邹先彪,男(汉族),博士,副主任医师.E-mail:xbzou@126.com
基金资助:
国家自然科学基金(39770002)

Amplified fragment length polymorphism analysis of Cryptococcus neoformans

ZOU Xian-biao¹, WEN Hai², LIAO Wan-qing², WU Jian-hua³, QIU Yun², YANG Jin-shui ⁴

1. Department of Dermatology, 304th Hospitol Affiliated to PLA General Hospital, Beijing 100048, China;
2. Department of Dermatology, Changzheng Hospitol, The Second Military Medical University, Shanghai 200003, China;
3. Department of Dermatology, Changhai Hospitol, The Second Military Mcrlical Universtiy, Shanghai 200433, China;
4. Insititute Genetic of Fudan Universtiy,Shanghai 200433, China;

Received:2010-02-11 Online:2010-04-28 Published:2010-04-28

摘要/Abstract

摘要： 目的评估AFLP-DNA指纹技术在新生隐球菌分类中应用情况。方法新生隐球菌基因组DNA用双酶酶切,双链接头连于其酶切末端,用与接头和酶切位点互补的引物扩增DNA片段,其产物在高分辨的变性聚丙酰胺凝胶上电泳分离,然后进行银染。结果分析来自5种血清型和临床分离株的18株新生隐球菌,可见有30多条大小在30～500bp的DNA-AFLP指纹,相同的血清型有不同的指纹图谱,来自同一患者不同病期的两株分离株和来自同一患者患者的不同部位的两株分离株都显示出相同的带型。结论显示了AFLP的高分辨率,是适用于新生隐球菌流行病学调查的有力工具。

关键词: 新生隐球菌, DNA, 扩增片段长度多态性

Abstract: Objective To evaluate AFLP-DNA fingerprinting method in the discrimination of C.neoformans.Methods Genomic DNA from C.neoformans was digested simultaneously with two restriction enzymes.Double-stranded adapters were ligated to the ends of restricted fragments and DNA fragments were amplified by PCR using primers complementary to the adapters and the restriction site sequences.AFLP products were separated using high resolution denaturing polyacrylamide gel electrophoresis and visualized by silver staining.Results DNA fingerprint patterns of 18 C.neoformans isolates from five serotypes and clinical isolates were analyzed by AFLP.AFLP fingerprints comprised over 30 bands detected in size range 30-500 bp.Different AFLP patterns were detected in isolates of the same serotype and identical banding patterns were observed from in isolates from the same patient at different time interals and different sources from one patiant.Conclusions AFLP is highly discriminatory,and powerful epidemiological tool for improving our understanding of C.neoformans.

Key words: Cryptococcus neoformans, DNA, AFLP

中图分类号:

R379.5

邹先彪, 温海, 廖万清, 吴建华, 仇芸, 杨金水. 新生隐球菌的AFLP分析[J]. 中国真菌学杂志, 2010, 5(2): 101-104.

ZOU Xian-biao, WEN Hai, LIAO Wan-qing, WU Jian-hua, QIU Yun, YANG Jin-shui. Amplified fragment length polymorphism analysis of Cryptococcus neoformans[J]. Chinese Journal of Mycology, 2010, 5(2): 101-104.

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