体外氟康唑诱导光滑念珠菌和近平滑念珠菌耐药及相关机制研究  

Induction of fluconazole resistance in Candida glabrata and Candida parapsilosis in vitro and corresponding mechanism

作　　者：刘锦燕[1] 王影 李文静[3] 赵悦[3] 孟玲宁 朱巍巍[1] 史册[3] 项明洁[1,3] 

机构地区：[1]上海交通大学医学院附属瑞金医院卢湾分院放免检验科,上海200020 [2]安徽省立医院南区检验科,安徽合肥230001 [3]上海交通大学医学院附属瑞金医院检验科,上海200025

出　　处：《诊断学理论与实践》2017年第5期510-515,共6页Journal of Diagnostics Concepts & Practice

基　　金：基金资助:上海市黄浦区优秀青年人才(RCPY1407);上海市医学重点专科(ZK2012A21)

摘　　要：目的:研究光滑念珠菌和近平滑念珠菌对氟康唑耐药的易感性及其诱导耐药株的耐药机制。方法 :选取对氟康唑敏感的光滑念珠菌、近平滑念珠菌临床株各1株以及对氟康唑敏感的光滑念珠菌标准株ATCC2001、近平滑念珠菌标准株ATCC22019,利用浓度递增法,在体外用氟康唑诱导使其成为耐药株,采用实时定量PCR检测外排泵相关基因、其转录因子和靶酶编码基因表达,并对光滑念珠菌PDR1转录因子、近平滑念珠菌MRR1转录因子和ERG11基因进行PCR扩增和测序。结果 :光滑念珠菌较近平滑念珠菌更易被氟康唑诱导为耐药株,CDR1的过度表达为其主要的耐药机制,对PDR1转录因子测序后发现了新的突变位点Y932C、V847F。近平滑念珠菌诱导耐药株的MDR1表达显著增加,其MRR1转录因子中亦存在新的突变位点P295S、I799S。结论 :光滑念珠菌和近平滑念珠菌对氟康唑的耐药易感性不同,耐药机制也存在差异。新发现的转录因子突变位点有待验证其功能。Objective： To investigate the susceptibility and mechanism of resistance to fluconazole in Candida glabrata and Candida parapsilosis isolates. Methods： Four strains （one clinical C. glabrata, one clinical C. parapsilosis, ATCC2001 and ATCC22019） were induced to be resistance strains in vitro by increasing concentrations of fluconazole. The expression levels of transporter genes and their transcription factor, and azole antifungal target gene were identified by real time-PCR. Meanwhile, the sequence of PDR1 gene of C. glabrata and MRR1, ERG11 gene of C. parapsilosis were de- termined by PCR based DNA sequencing. Results： C. glabrata was more prone to be resistant to fluconazole than C. parapsilosis. CDR1 was significantly overexpressed in resistant strains of C. glabrata, while MDR1 was significantly overex- pressed in resistant strains of C. parapsilosis. PDR1 mutations Y932C, V847F were identified in resistant strains of C. glabrata, and MRR1 mutations P295S, I799S were found in resistant strains of C. parapsilosis. Conclusions： C. glabrata and C. parapsilosis showed different susceptibility to fluconazole resistance, and difference also occurred in resistance mechanism. And the function of new identified mutations in transcription factor needs further verification.

关 键 词：念珠菌 氟康唑耐药 外排泵 靶酶 转录因子 

分 类 号：R519[医药卫生—内科学]