作　　者： (张甜甜）; (朱玉霞）; (樊尚荣）; (石玉）;

机构地区： 安徽医科大学北京大学深圳医院临床学院,深圳518036 北京大学深圳医院,深圳518036

出　　处： 《现代妇产科进展》 2017年第8期564-568,573,共6页

摘　　要： 目的:应用多位点序列分型(MLST)分析38株热带假丝酵母菌的亲缘关系,体外比较热带假丝酵母菌对常见抗真菌药物的敏感性,检测耐药基因的表达水平。方法:通过对6对等位基因进行测序,用e BURST分组和非加权配对算术平均法(UPGMA)聚类分析菌株的进化和变异。微量肉汤稀释法进行体外抗真菌药物敏感试验,实时荧光定量聚合酶链反应(q-RT-PCR)检测耐药基因CDR1、CDR2、ERG1、ERG3、ERG6、ERG9和ERG11表达。结果:38株菌株中获得30个二倍体序列(DST),其中14个新DST,11个新等位基因,27个DST为单体型。e BURST分组,主要分成2组,大多数DST属于单体型。聚类分析总共分18个进化枝。阿尼芬净、米卡芬净、卡泊芬净、氟康唑、伏立康唑、伊曲康唑、两性霉素B、氟胞嘧啶、咪康唑、布康唑、克霉唑、特康唑、特比萘芬和制霉菌素14种抗真菌药物的MIC90分别为0.030μg/ml、0.015μg/ml、0.500μg/ml、1.000μg/ml、0.125μg/ml、0.250μg/ml、1.000μg/ml、0.250μg/ml、2.000μg/ml、2.000μg/ml、0.250μg/ml、0.250μg/ml、256.000μg/ml和8.000μg/ml。绝大多数耐药基因表达量较高的菌株可能在进化枝11和14。结论:热带假丝酵母菌MLST变异较大,菌株之间遗传距离较远。热带假丝酵母菌对特比萘芬普遍耐药,对目前其他常用的抗真菌药物多为敏感。 Objective： To analyze the relationship between 38 strains of C. tropicalis from vulvovaginal candidasis by using the multilocus sequence typing（ MLST） method. To compare the susceptibility of C.tropicalis to common antifungal agents in vitro and the expression of common resistance genes in C.Tropicalis.Methods： We sequenced the 6 alleles of C.tropicalis,compared them with C.tropicalis MLST database and obtained diploid sequence types（ DST）.We grouped the DST by using e BRUST and constructed the phylogenetic tree by using unweighted pair group method with arithmetic averages（ UPGMA） method.Antifungal susceptibility test was detected in 38 strains of C.tropicalis by using broth dilution method.The expression of CDR1,CDR2,ERG1,ERG3,ERG6,ERG9 and ERG11 were detected by using real-time fluorescent quantitative polymerase chain reaction（ q-RT-PCR）. Results： We obtained 30 DSTs and 18 clades from 38 strains of C.tropicalis,of which 14 were new DSTs,11 were new alleles and 27 DSTs were singleton. The minimum inhibitory concentration 90（ MIC90） of anidulafungin,micafungin,caspofungin,fluconazole,voriconazole,itraconazole,amphotericin B,flucytosine,miconazole,butoconazole,clotrimazole,terconazole,terbinafine and nystatin for C.tropicalis were： 0. 030μg/ml,0. 015μg/ml,0. 500μg/ml,1. 000μg/ml,0. 125μg/ml,0. 250μg/ml,1.000μg/ml,0.250μg/ml,2. 000μg/ml,2. 000μg/ml,0. 250μg/ml,0. 250μg/ml,256.000 μg/ml and 8.000μg/ml,respectively.The antifungal resistance genes expression levels in clade 11 and clade 14 strains were higher than in the others.Conclusion： The variation of C.tropicalis based on MLST was larger.C.tropicalis was susceptible to commonly used antifungal agents.

关 键 词： 外阴阴道假丝酵母菌病 热带假丝酵母菌 多位点序列分型 药物敏感性 耐药基因