烟曲霉伊曲康唑耐药株的体外诱导及与CYP 51A基因相关性研究.pdfVIP

Journal of Practical Dermatology, Aug. ,Vol. No.4 实用皮肤病学杂志 2014年8月第7卷第4期 2014 7 253 烟曲霉伊曲康唑耐药株的体外诱导 及与CYP 51A基因相关性研究 敖俊红，廖　勇，郝震锋，祝　贺，杨蓉娅 [摘要]　 目的 构建烟曲霉体外诱导伊曲康唑耐药株，初步探讨与 CYP 51A 基因的相关性。 方法　通过含有伊曲康唑的固体培养基对烟曲霉进行梯度浓度体外诱导耐药，微量液基稀释 法测定诱导株对伊曲康唑的敏感性；提取诱导株基因组，扩增唑类药物靶酶 CYP 51A 基因， 进行测序和序列分析。结果　诱导获得 8 株烟曲霉体外诱导株，其中 6 株对伊曲康唑表现出 耐药性。耐药株唑类药物靶酶 CYP 51A 基因发生 G54 (G54R, G54E) 和 L98H 的点突变。结 论　伊曲康唑体外梯度浓度诱导是获得烟曲霉伊曲康唑耐药株的有效方法；烟曲霉对伊曲康 唑抗真菌药物的耐药性可能与唑类药物靶点基因 CYP 51A 的点突变相关。 [ ] 关键词 　 烟曲霉；伊曲康唑；耐药，体外诱导；CYP 51A基因 [ ] R379.6 R978.5 [ ] A [ ]　 1674-1293(2014)04-0253-03 中图分类号　 ； 　　 文献标识码 　　 文章编号 In vitro evolution of itraconazole resistance in Aspergillus fumigatus and its relationship with CYP 51A gene AO Jun-hong ，LIAO Yong，HAO Zhen-feng，et al Institute of Skin Damage and Repair, General Hospital of Beijing Military Command, Beijing 100700, China [Abstract]　Objective　To obtain in vitro itraconazole-resistant strains of Aspergillus fumigatus and explore its relationship with CYP 51A gene. Methods　Wild type strain of Aspergillus fumigatus was serially transferred to agar medium containing itraconazole of gradually increased concentration in vitro. The in vitro susceptibility of induced strains was determined by broth microdilution assay. The genomes of itraconazole-resistant mutant strains were extracted, then the azole drugs target enzyme gene CYP 51A was amplified, sequenced and analyzed. Results　Eight induced strains of Aspergillus fumigatus in vitro were obtained, and six strains shown resistance to itraconazole. Azole drugs target enzyme gene CYP 51A of itraconazole-resistant strains occurs G54 (G54R, G54E) and L98H point mutations. Conclusions　Itraconazole-resistant strains of Aspergillus fumigatus can be induced in vitro by concentration gradient. Point mutations of azole antifungal drug target enzyme ge