耐氟康唑阴道白色念珠菌ERG基因家族突变研究及唑类药物抑菌分析  

Study on ERG gene family mutation in fluconazole-resistant vaginal Candida albicans and antimicrobial analysis of Azole drugs

作　　者：李倩珺[1] 黄淑洪 冯泳琳 林卫虹[1] Qianjun Li;Shuhong Huang;Yonglin Feng;Weihong Lin(The First Affiliated Hospital of Guangzhou Medical University,Guangzhou Guangdong 510120,China;KingMed School of Laboratory Medicine,Guangzhou Medical University,Guangzhou Guangdong 510182,China)

机构地区：[1]广州医科大学附属第一医院检验科,广东广州510120 [2]广州医科大学金域检验学院,广东广州510182

出　　处：《中华临床实验室管理电子杂志》2022年第3期137-141,152,共6页Chinese Journal of Clinical Laboratory Management(Electronic Edition）

基　　金：广东省中医药局中医药科研项目(20211241)。

摘　　要：目的研究耐氟康唑阴道白色念珠菌ERG11、ERG3和ERG5基因突变,及分离株对酮康唑、克霉唑、硝酸咪康唑的抑菌分析。方法收集221株经质谱仪鉴定为白色念珠菌的菌株,微量肉汤稀释法进行氟康唑、酮康唑、克霉唑、硝酸咪康唑药敏试验,得出四种药物对白色念珠菌最小抑菌浓度(MIC)值;根据CLSI M27-A3筛选出氟康唑非敏感白色念珠菌,提取DNA后扩增ERG11、ERG3与ERG5基因,进行双向测序,与GenBank中的标准菌株基因序列(SC5314和AF069752)进行BLAST比对分析。结果白色念珠菌对氟康唑耐药率为9.05%;对白色念珠菌MIC值进行比较,酮康唑的MIC值均低于克霉唑和硝酸咪康唑,差异有统计学意义(P<0.05)。克霉唑与硝酸咪康唑比较,差异无统计学意义(P>0.05)。ERG11、ERG3、ERG5分别发现18、17、12种错义突变。结论酮康唑对白色念珠菌抑菌率总体比克霉唑、硝酸咪康唑强。耐氟康唑白色念珠菌ERG11、ERG3、ERG5基因出现若干错义突变位点,某些位点突变导致氨基酸变异可能与耐药产生有关。Objective To study ERG11,ERG3 and ERG5 gene mutations of Fluconazol-resistant vaginal Candida albicans and the antimicrobial analysis of the isolates against ketoconazole,clotrimazole and miconazole nitrate.Methods 221 strains of Candida albicans were collected and identified by mass spectrometer.Fluconazole,ketoconazole,clotrimazole and miconazole nitrate were tested by microbroth dilution method,and the minimum inhibitory concentration(MIC)of the four drugs against Candida albicans was obtained.Fluconazole non-sensitive Candida albicans was screened according to CLSI-M27.ERG11,ERG3 and ERG5 genes were amplified after DNA extraction,and bidirectional sequencing was performed,which was compared with the standard sequence(SC5314)and AF069752 in GenBank.Results The resistance rate of Candida albicans to fluconazole was 9.05%.Compared with the MIC values of Candida albicans,the MIC values of ketoconazole were lower than those of clotrimazole and miconazole nitrate,with statistical significance(P<0.05).There was no significant difference between clotrimazole and miconazole nitrate(P>0.05).ERG11,ERG3 and ERG5 were showed 18,17 and 12 missense mutations,respectively.Conclusion Ketoconazole has stronger antibacterial effect on Candida albicans than metrimazole and miconazole nitrate..Several missense mutations occurred in ERG11,ERG3 and ERG5 genes of Fluconazol-resistant Candida albicans,and amino acid variation caused by mutations at some sites may be related to drug resistance.

关 键 词：白色念珠菌 ERG基因 氟康唑 耐药 

分 类 号：R446.5[医药卫生—诊断学]