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目的探讨临床分离白色念珠菌ERG5基因突变、高表达与唑类抗真菌药物耐药的关系。方法对36株临床白色念珠菌进行药物敏感性试验,并对其ERG5基因进行扩增测序,将测序结果与Gen Bank中已发表的序列(Gen Bank序列号:AF069572)进行比对,分析基因突变情况;抽提白色念珠菌临床菌株的总RNA,并反转录合成c DNA,采用实时荧光定量PCR(FQ-RT-PCR)方法检测ERG5基因的表达水平。结果 36株白色念珠菌临床菌株中,共检测出ERG5基因存在1个错义突变位点;氟康唑耐药组有11株发生ERG5高表达,占61.11%,敏感组中5株发生高表达,占31.25%。耐药菌株组ERG5基因的表达量ΔCt值与敏感组比较,差异具有统计学意义(P<0.05)。结论白色念珠菌ERG5基因的高表达可能与FCA产生耐药有关,并未发现ERG5耐药菌株在同一位点发生错义突变,白色念珠菌ERG5基因突变、高表达在耐药中的作用还有待于进一步研究证实。
Objective To investigate the relationship between the clinical isolates of Candida albicans ERG5 gene mutation and high level of resistance to azole antifungal agents. Methods 36 strains of clinical Candida albicans were tested for drug susceptibility and the ERG5 gene was amplified and sequenced. The results of sequencing were compared with those published in Gen Bank (Gen Bank serial number: AF069572) The total RNA was extracted from Candida albicans clinical strains and reverse transcribed into c DNA. The expression of ERG5 gene was detected by real-time fluorescence quantitative PCR (FQ-RT-PCR). Results A total of 36 strains of Candida albicans were found in the clinical isolates of Candida albicans. One missense mutation site was found in the ERG5 gene. Eleven of the fluconazole-resistant groups showed high expression of ERG5 (61.11%), and five of the sensitive strains were highly expressed , Accounting for 31.25%. The expression of ΔCt in ERG5-resistant strains was significantly different from that in the sensitive groups (P <0.05). Conclusion The high expression of Candida albicans ERG5 gene may be related to the development of FCA resistance. No ERG5-resistant strains were found to have the missense mutation at the same site. The role of Candida albicans ERG5 gene mutation and high expression in drug resistance remains to be determined In further study confirmed.