上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（临床研究） • 上一篇    下一篇

耐氟康唑白假丝酵母靶酶和外排泵编码基因过表达检测

史 册1,2，刘锦燕2，魏 冰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， 上海 200025； 2.卢湾分院放免检验科， 上海 200020
  • 出版日期:2013-08-28 发布日期:2013-09-16
  • 通讯作者: 项明洁, 电子信箱: mjxiang123456@126.com。
  • 作者简介:史 册（1990—）， 男， 硕士生; 电子信箱: shice1602@163.com。
  • 基金资助:

    上海市科委基金（114119b0500）和上海市卫生局基金（2009239）

Detection of overexpression of genes encoding target enzyme and efflux pumps in fluconazole-resistant Candida albicans

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

  1. 1.Department of Clinical Microbiology Laboratory, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai200025, China; 2.Radioimmunology and Clinical Laboratory, Luwan Branch, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200020, China
  • Online:2013-08-28 Published:2013-09-16
  • Supported by:

    Shanghai Science and Technology Committee Foundation, 114119b0500; Shanghai Municipal Health Bureau Foundation, 2009239

PDF (PC)

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摘要：

目的 研究临床氟康唑耐药白假丝酵母中耐药相关基因MDR1、CDR1、CDR2和ERG11的表达水平及其与耐药的关系。方法 采用Real-Time PCR技术检测耐药相关基因MDR1、CDR1、CDR2和ERG11的表达水平，计算敏感组菌株基因△Ct的平均值与标准差，低于－3 s为过表达。罗丹明6G (Rhodamine6G)外排实验检测外排泵高表达的白假丝酵母菌株中的外排功能。结果 在12株耐药株中，4个基因中以CDR1和CDR2表达上调最为显著，分别有8株和3株，而ERG11只有1株表达上调，未见MDR1表达上调。外排实验中外排泵表达升高的菌株外排能力强，加入葡萄糖之后有些菌株外排进一步增强，而敏感菌株加入葡萄糖之前几乎无外排，加入葡萄糖之后略有外排。结论 本实验基因表达与外排泵功能检测结果均证实，白假丝酵母对氟康唑耐药与ABC转运蛋白超家族过表达有关。

关键词: 白假丝酵母, 氟康唑, 耐药机制, 靶酶, 外排泵, 过表达

Abstract:

Objective To investigate the expression of MDR1, CDR1, CDR2 and ERG11 genes in clinical fluconazole-resistant Candida albicans isolates, and explore its relationship with drug resistance. Methods The expression of drug resistance related genes MDR1, CDR1, CDR2 and ERG11 was detected by Real-Time PCR, and mean and standard deviation of △Ct in genes of susceptible strains were calculated, with lower than －3 s as overexpression. The efflux of Rhodamine6G was determined to examine the function of efflux pumps in the overexpression strains of Candida albicans. Results Among the 12 drug resistant strains, CDR1 and CDR2 were the most notable genes, with overexpression in 8 and 3 strains respectively. There was one strain overexpressing ERG11, and none for MDR1. The efflux of Rhodamine6G increased in the strains overexpressing efflux pumps, and the effect was enhanced after addition of glucose. As to the susceptible strains, there was almost no efflux before addition of glucose, and there was a slight change after addition of glucose. Conclusion Fluconazole-resistance is related to the overexpression of ABC transport protein superfamily, which is confirmed by the gene expression profile and function test of efflux pumps.

Key words: Candida albicans, fluconazole, mechanism of resistance, target enzyme, efflux pump, overexpression