烟曲霉的微管蛋白基因鉴定及唑类抗真菌药物的耐药性研究

目的建立烟曲霉临床分离菌株的微管蛋白基因鉴定方法,并分析烟曲霉对唑类抗真菌药物的耐药性。方法对表型鉴定为烟曲霉的菌株,采用十六烷基三甲基溴化铵(CTAB)方法提取烟曲霉菌株DNA,并采用对微管蛋白基因进行PCR扩增和测序的方法对烟曲霉菌株进行分子生物学鉴定,采用琼脂稀释法检测烟曲霉菌株对伊曲康唑和伏立康唑的耐药性。结果 91株临床分离株表型鉴定为烟曲霉,这些菌株在35℃和48℃生长。其中88株(96.70%)经微管蛋白基因鉴定为烟曲霉。87株烟曲霉菌株对4μg/mL伊曲康唑和1μg/mL伏立康唑敏感,检出1株对4μg/mL伊曲康唑和1μg/mL伏立康唑耐药菌株。结论微管蛋白基因鉴定方法可以对烟曲霉进行准确鉴定。

Identification of Aspergillus fumigatus through tubulin gene amplification and drug resistance to antifungal azoles agents

Objective To identify Aspergillus fumigatus through method of tubulin gene amplification and analyse drug resist‐ance to azole antifungal agents .Methods By using cetyltrimethyl ammonium bromide (CTAB) method ,genomic DNA from asper‐gillus fumigatus were extracted .Tubulin gene was amplified by polymerase chain reaction amplification assay and series analysis . Drug resistance of aspergillus fumigatus to antifungal azoles agents were determined by agar dilution method .Results A toal of 91 strains separated from specimen were identified as aspergillus fumigatus through phenotypic identification ,which grew at 35 ℃and 48 ℃ .Through tubulin gene amplification ,88 (96 .70% ) of the 91 strains were confirmed as aspergillus fumigatus .And 87 strains of the 88 Aspergillus fumigatus were confirmed sensitive to 4 μg/mL itraconazole and 1 μg/mL voriconazole ,and 1 strain was re‐sistant to 4 μg/mL itraconazole and 1 μg/mL voriconazole .Conclusion Aspergillus fumigatus could be identified by method of tu‐bulin gene amplification accurately .