多药耐药外排泵ABC基因对白色假丝酵母菌及克柔假丝酵母菌耐药性产生的研究

发布时间：2018-11-12 17:00

【摘要】：目的探讨多药耐药外排泵基因在白色假丝酵母菌和克柔假丝酵母菌耐唑类药物中的作用,对假丝酵母菌属的耐药研究提供理论基础。方法通过体外药敏试验筛选出临床白色假丝酵母菌和克柔假丝酵母菌对伊曲康唑的耐药菌株和敏感菌株,使用实时荧光定量PCR(FQ-RT-PCR)方法检测耐药和敏感菌株多药耐药外排泵基因(CDR1/CDR2/ABC1)的mRNA表达水平,使用罗丹明6G为示踪剂了解白色假丝酵母菌和克柔假丝酵母菌的药物外排情况。结果临床分离的白色假丝酵母菌和克柔假丝酵母菌对氟胞嘧啶、两性霉素B的敏感率均较高,均80.00%,白色假丝酵母菌对伊曲康唑的耐药率较高,70.00%;根据实时荧光定量RT-PCR实验获得的ΔCt值,经统计分析发现,白色假丝酵母菌耐药菌株CDR1、CDR2mRNA的表达量高于敏感菌株的表达量,差异有统计学意义(P0.05),但克柔假丝酵母菌中仅有一株耐药菌株出现高表达,敏感菌株和耐药菌株ABC1mRNA的表达量差异无统计学意义;通过罗丹明6G外排试验测定,随时间延长罗丹明6G外排明显增加,在CDR1/CDR2mRNA高表达的耐药菌株明显增高。结论外排泵基因CDR1、CDR2在白色假丝酵母菌耐药性的产生中可能起到一定作用,但不能说明外排泵基因ABC1在克柔假丝酵母菌耐药性的产生中无作用,因样本量较小,需要扩大样本量进一步研究。
[Abstract]:Objective to investigate the role of multidrug resistant efflux pump gene in the study of resistance to azoles in Candida albicans and Candida krrostris, and to provide a theoretical basis for the study of drug resistance of Candida. Methods the resistant and sensitive strains of clinical Candida albicans and Candida krorostris to itraconazole were screened by drug sensitivity test in vitro. Real-time fluorescence quantitative PCR (FQ-RT-PCR) was used to detect the mRNA expression of multidrug resistant efflux pump gene (CDR1/CDR2/ABC1) in drug-resistant and sensitive strains. Rhodamine 6G was used as tracer to understand the drug efflux of Candida albicans and Candida krorostris. Results the sensitivity rates of Candida albicans and Candida krorostris to fluorocytosine and amphotericin B were both high (80.00g). The resistance rate of Candida albicans to itraconazole was high (70.00g). According to the 螖 Ct value obtained by real-time fluorescence quantitative RT-PCR test, the expression of CDR1,CDR2mRNA in Candida albicans resistant strain was higher than that of sensitive strain (P0.05). However, only one resistant strain of Candida krorostris showed high expression, and there was no significant difference in the expression of ABC1mRNA between sensitive strain and resistant strain. The results of Rhodamine 6G efflux test showed that the efflux of Rhodamine 6G increased with time, and that of drug-resistant strains with high expression of CDR1/CDR2mRNA increased significantly. Conclusion the efflux pump gene CDR1,CDR2 may play a certain role in the production of resistance of Candida albicans, but it does not indicate that the efflux pump gene ABC1 has no effect on the production of resistance of Candida krrostris because the sample size is small. Further research is needed to expand the sample size.
【作者单位】：山西医科大学第二医院皮肤科;
【基金】：山西省青年科技研究基金资助项目(2012021035-3) 山西省基础研究基金资助项目(2012011045-3) 山西省高等学校科技公关基金资助项目(20141106) 山西省卫生厅科技攻关基金资助项目(201301009) 山西医科大学科技创新基金资助项目(01201013);山西医科大学第二医院青年基金资助项目(20110104)
【分类号】：R440

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