白念珠菌临床株氟康唑敏感性试验与ERG11基因突变检测.pdfVIP

756 1 Q鲞 1 期 Chin』 ennatol，December 2007．V01．40。N0．12 · 论 著 · 白念珠菌临床株氟康唑敏感性试验 与ERG1 1基因突变检测 徐永豪 李春阳 陈腊梅 胡勤峰 【摘要】目的 了解白念珠菌临床株对氟康唑的敏感性，筛查临床菌株的ERG11突变，探讨其与耐 药的关系。方法 收集鉴定白念珠菌临床株，采用微量稀释法和药敏纸片法体外测定试验菌株对氟康唑 的敏感性。分段扩增ERG11基因的第295～777 bp(483 bp)、第723～1204 bp(482 bp)和第1179～ 1667 bp(489 bp)等3个片段，并测序。结果 共收集念珠菌80株，分离白念珠菌52株，52株白念珠菌 均为敏感株。经测序的5株敏感白念珠菌ERG11基因共出现l9处突变，其中l4处为同义突变，其余 5处错义突变(T495A、A530C、G640A、A945C、G1609A)分别可引起羊毛甾醇l4 一去甲基酶的下列氨基酸 改变：D116E、K128T、E165K、E266D、V488I。结论 白念珠菌临床株对氟康唑敏感性较高；E165K和 V488I可能与耐药无关。 【关键词】念珠菌，白色；微生物敏感性试验；氟康唑；突变；ERG11；羊毛甾醇l4 一去甲基酶 Correlation beb’reen anfifungal susceptibility of clinical isolates of Candida albieans to fluconazole and ERGl1 mutations XU Yong-hao．LI Chun-yang,CHEN La—me／,HU Q／n-feng．Department ofDer- matology,Qilu Hospital,Shandong University,Jinan 250012,China Corresponding author：LI Chun-yang,email：lichunyang@medmail．corn．cn 【Abstract】 Objective To analyze the sensitivity of clinicalisolates of Candida albicans to flucona— zole．to detect mutations in their ERGl1 genes．and to investigate the correlation between ERGl1 gene mutation and resistance to fluconazole．Methods Candida albicans was identified from clinical isolates of Candida spp—The sensitivity to fluconazole was detected抽 v／no by microdilution—basesd method and Rosco tablets method．Three pairs of primers were designed to amplify three fragments of ERG1 l gene (483 bps， from 295 bp to777 bp；482bps，from 723 bp to 1204 bp；489 bps，from l179 bp to 1667 bp)after the extracting of genomic DNA．PCR proIcts were sequenced．Results Eighty clinical isolates of Candida spp．were collected．which included 52 isolates of Clan da albicans．all of which were sensitive to flucona— zole．Nineteen mutations were detected in ERGll gene of 5 fluconazole．sensitive clinical isolates．Of the 19 mutations，14