摘要
目的探讨白念珠菌锌簇转录因子Cap1、Mrr1基因突变/高表达及外排泵基因MDR1的mRNA表达水平在唑类抗真菌药物氟康唑(FCA)、伊曲康唑(ITR)、伏立康唑(VRC)交叉耐药中的作用。方法收集山西医科大学第二医院皮肤性病科门诊疑似外阴阴道念珠菌病(VVC)患者阴道分泌物标本,进行真菌直接镜检、培养、分离鉴定和体外药物敏感性试验,获得试验用白念珠菌耐药和敏感菌株共68株。采用PCR方法扩增68株白念珠菌临床分离菌株Cap1、Mrr1基因,并进行双向测序,应用Blast软件,将测序结果与GenBank中已发表的序列进行比对,分析基因突变情况,实时荧光定量PCR检测锌簇转录因子Cap1、Mrr1和外排泵基因MDR1的mRNA表达水平。结果Cap1测序中1株ITR、VRC交叉耐药菌株出现P311S错义突变。Mrr1测序中1株FCA、ITR、VRC交叉耐药菌株出现5个错义突变(T917M,I'923I,E1020Q,F1032L,S1037L);FCA、ITR、VRC交叉耐药菌株组中Cap1/Mrr1/MDR1的mRNA表达量分别高于敏感菌株组;CA、ITR、VRC交叉耐药菌株组Cap1和MDR1(r=0.173,P=0.571),Mrr1和MDR1(r=-0.091,P=0.766),Cap1和Mrr1基因(r=0.025,P=0.936)的mRNA表达量不相关(均P〉0.05)。结论Cap1和Mrr1的突变可能和唑类药物交叉耐药有关。
Objective To investigate the role of zinc cluster transcription factors Capl and Mrrl mutation/high expression and the mRNA level of efflux pump MDR1 in Candida albicans cross-resistant to azole antifungal agents such as fluconazole (FCA) , itraeonazole (ITR) , and vorieonazole (VRC). Methods Vaginal swabs from patients with suspected vulvovaginal candidiasis (VVC) in the Department of Dermatology, Second Hospital of Shanxi Medical University, were collected for direct microscopy, culture, isolation, identification and in vitro drug sensitivity test. A total of 68 Candida albicans resistant and susceptible strains were obtained. Capl and Mrrl in the 68 clinically isolated strains of Candida albicans were PCR- amplified and bidirectionally sequenced. Blast software was used to compare the sequencing results with published sequences in GenBank for identification of gene mutations. Real- time fluorescent quantitative PCR was used to detect mRNA expression of zinc cluster transcription factors Cap 1 and Mrrl and efflux pump gene MDR1. Results By Cap l sequencing, P311S missense mutation was found in one strain cross-resistant to ITR and VRC. By Mrrl sequencing, five missense mutations (T917M, T923I, E1020Q, F1032L, and S1037L) were found in one strain cross-resistant to FCA, ITR and VRC. The expression level of Capl/M1T1/MDR1 mRNA was higher in FCA, ITR and VRC cross-resistant strains than in sensitive strains. In CA, ITR and VRC cross-resistant strains, there was no correlation in expression levels between Capl and MDR1 (r=0.173, P=0.571), between Mrrl and MDR1 (r=-0.091, P=0.766), or between Capl and Mrr1 (r=0.025, P=0.936) (all P〉0.05). Conclusion This study showed that mutations in Capl and Mrrl may be related to azoles cross-resistance.
作者
杨静
冯文莉
杨璐
祝欣
奚志琴
乔祖莎
岑雯
Yang Jing;Feng Wenli;Yang Lu;Zhu Xin;Xi Zhiqin;Qiao Zusa;Cen Wen(Department of Dermatology,Second Affiliated Hospital of Shanxi Medical University,Taiyuan 030001,Shanxi,China)
出处
《中华生物医学工程杂志》
CAS
2017年第5期353-362,共10页
Chinese Journal of Biomedical Engineering
基金
山西省应用基础研究计划(201701D121171)
山西省重点研发计划(指南)项目(201603D321063)
山西省卫生和计划生育委员会科研课题(201601050)
关键词
念珠菌病
外阴阴道
念珠菌
白色
转录因子
外排泵
基因突变
Candidiasis
vulvovaginal
Candida albicans
Transcription factor
Efflux pump
Gene mutation