上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

重复序列PCR与多位点分型技术在白假丝酵母菌基因分型中的比较

魏 冰1,2，刘锦燕2，史 册1，项明洁1,2   

  1. 上海交通大学 医学院附属瑞金医院 1.检验科； 2.卢湾分院放射免疫检验科， 上海 200025
  • 出版日期:2013-11-28 发布日期:2013-12-03
  • 通讯作者: 项明洁, 电子信箱: mjxiang123456@126.com。
  • 作者简介:魏 冰（1986—）， 女， 技师， 硕士生; 电子信箱: weibing1207@126.com。
  • 基金资助:

    上海市卫生局资助项目（2009239）；上海市科委生物医药资助项目（11411960500）；上海市医学重点专科建设项目（2012年A类21号）

Comparative study on genotyping of Candida albicans by repetitive sequence-based PCR and multilocus sequence typing

WEI Bing1,2, LIU Jin-yan2, SHI Ce1, XIANG Ming-jie1,2   

  1. 1.Clinical Laboratory; 2.Radioimmunology and Clinical Laboratory Luwan Branch, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China
  • Online:2013-11-28 Published:2013-12-03
  • Supported by:

    Shanghai Municipal Health Bureau Foundation, China, 2009239; Bio-medical Program of Science and Technology Commission of Shanghai Municipality, China, 11411960500; Key Medical Disciplines Construction Program of Shanghai, China, 2012-A-21

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摘要：

目的 应用多位点序列分型技术（MLST）和重复序列聚合酶链反应（REP-PCR）对临床分离出的白假丝酵母菌进行分型，并对两种方法作出应用评价。方法 收集上海地区医院中真菌性阴道炎分离出的40株白假丝酵母菌，选择合适引物进行扩增，通过电泳比较分析获得REP-PCR分型。选择7对管家基因进行PCR反应并测序，将测序结果与标准序列进行比对，获得由7对管家基因组成的等位基因谱，最终得到相应的序列分型。结果 REP-PCR中Ca22-Ca22引物对得到电泳条带最优，40株白假丝酵母菌共分成7种REP-PCR型；MLST分型得出29种，且均为新型。结论 MLST分辨率较REP-PCR高，但由于REP-PCR分型方法更为快捷、经济，更适用于实验室大量菌株分型和临床分型。MLST则更客观，更适合用于进化学的研究及全球性流行病学的调查研究。

关键词: 白假丝酵母菌, 多位点序列分型技术, 重复序列聚合酶链反应, 基因分型

Abstract:

Objective To compare repetitive sequence-based polymerase chain reaction (REP-PCR) and multilocus sequence typing (MLST) in genotyping of Candida albicans, and assess the two methods. Methods Forty Candida albicans strains were isolated from Candida vaginitis patients from local hospitals of shanghai. With the most suitable primers used to amplify, the amplicons were analyzed through electrophoresis to generate different REP-PCR types. Seven loci in housekeeping genes were amplified and sequenced. The sequences were compared with MLST database to obtain sequence type (ST). Results For REP-PCR, Ca22-Ca22 has the best genotyping effect. Seven REP-PCR types were found in 40 Candida albicans isolates. Twenty-nine MLST types were found and they were all new types. Conclusion MLST has much better discriminatory power than REP-PCR, but REP-PCR offers a simple and rapid method for molecular typing. Therefore, REP-PCR can be the first choice for a large number of isolates in laboratory and clinical practice. MLST is more objective and more suitable for the research of evolutionary and global epidemiology investigation.

Key words: Candida albicans, multilocus sequence typing, repetitive sequence-based polymerase chain reaction, genotyping