摘要
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 of over- expression and mutation of ERG5 with the drug resistance of anti fungal drugs in the clinical isolates of C. albicans. Methods The drug susceptibility tests were conducted on 36 clinical isolates of C. albicans. Then all the ERG5 genes were measured and amplified,and the sequences results were compared to a known ERG5 sequence in Gen Bank（ AF069572） to identify the mutation of ERG5. Total RNA of clinical isolates of Candida albicans was extracted,and c DNA was synthesized by reverse transcription. The expression level of ERG5 gene was detected by real- time fluorescence quantitative PCR（ FQ- RT- PCR）. Results There was one mutation of ERG5 identified from the 36 isolates. The over expression of ERG5 was identified on 11 strains in FCA- resistant group,accounting for 61. 11%,and on 5strains in FCA- susceptible group,accounting for 31. 25%. Compared with in the sensitive group,there was difference on the expression volume of ERG5（ ΔCt） in the drug resistant group,and it was with statistical significance（ P〈0. 05）. Conclusion This experiment suggested that the over expression of ERG5 was positively correlated to the FCA- resistant in C. Albicans,and we had not found missense mutation in the same site. The role for mutation and elevated ERG5 gene expression to drug resistance in Candida albicans was not yet verified.
出处
《中国卫生检验杂志》
CAS
2016年第4期542-545,共4页
Chinese Journal of Health Laboratory Technology
基金
山西省青年科技研究基金(2012021035-3)
山西省基础研究项目(2012011045-3)
山西省高等学校科技公关项目(2014)
山西省卫生厅科技攻关项目(20130-1009)
山西医科大学科技创新基金(01201013)
山西医科大学第二医院青年基金(20110104)
