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目的探究白念珠菌生物膜形成过程中不同时期的锌簇转录因子TAC1与外排泵基因MDR1,CDR1,CDR2与白念珠菌生物膜耐药的关系。方法 96孔板培养白念珠菌生物膜,MTT法分别测定白念珠菌生物膜形成不同时期(4h,24h,48h)对氟康唑的敏感性。6孔板培养白念珠菌生物膜,分别在4h,24h,48h提取生物膜的总RNA,逆转录cDNA,RT-PCR检测TAC1,MDR1,CDR1,CDR2的基因表达量。结果白念珠菌在生物膜的形成过程中TAC1,MDR1,CDR1,CDR2的基因表达差异具统计学意义,经过Bonferroni多重比较,结果显示标准株00889 TAC1,MDR1,CDR1,CDR2早期基因表达量比中、晚期高。临床敏感株359 TAC1,MDR1,CDR1,CDR2中期基因表达量比晚期高。结论锌簇转录因子TAC1和外排泵基因可能在白念珠菌生物膜的早中期对氟康唑的耐药形成中发挥了一定作用。
Objective To investigate the relationship between zinc cluster transcription factor TAC1 and efflux pump genes MDR1, CDR1 and CDR2 and the resistance to Candida albicans biofilm during different stages of C. albicans biofilm formation. Methods 96-well plate culture Candida albicans biofilms, MTT assay were determined at different stages of Candida albicans biofilm formation (4h, 24h, 48h) susceptibility to fluconazole. The biofilm of C. albicans was cultured in 6-well plate. The total RNA of biofilm was extracted at 4h, 24h and 48h respectively. The mRNA of TAC1, MDR1, CDR1 and CDR2 was detected by RT-PCR. Results The gene expression of TAC1, MDR1, CDR1 and CDR2 of Candida albicans in biofilm formation was statistically significant. After Bonferroni multiple comparisons, the results showed that the early gene expression of TAC1, MDR1, , Late high. The expression of metaphase gene in clinical sensitive strains 359 TAC1, MDR1, CDR1 and CDR2 was higher than that in the late stage. Conclusion The genes of zinc cluster transcription factor TAC1 and efflux pump may play a role in fluconazole resistance formation in early and middle period of Candida albicans biofilm.