ITS和β-微管蛋白基因序列鉴定曲霉菌的临床应用  

The clinical application of identification of Aspergillus spp.using ITS and β-tubulin gene sequences

作　　者：朱敏[1] 耿佳靖[1] 王玫[1] 黄艳飞[1] 王利君[1] 鲁辛辛[1] 

机构地区：[1]首都医科大学附属北京同仁医院检验科,100730

出　　处：《中华检验医学杂志》2011年第4期353-357,共5页Chinese Journal of Laboratory Medicine

基　　金：国家自然科学基金资助项目（30772066）

摘　　要：目的 研究ITS序列分析和β-微管蛋白基因序列分析在曲霉菌鉴定中的临床应用价值.方法 收集2007年7月至2010年1月,首都医科大学附属北京同仁医院真菌性鼻窦炎病原菌株中曲霉菌124株,分别对其进行形态学和分子鉴定.形态学包括传统培养方法、玻片培养和乳酸酚棉蓝染色及KOH消化后显微镜镜检.将菌株的PCR扩增产物进行ITS序列分析和β-微管蛋白基因序列分析,其测序结果与GenBank、European Molecular Biology Laboratory和DNA Data Bank of Japan 3个数据库比对,得到分子鉴定结果.结果 形态学鉴定为黄曲霉的56株曲霉中,经ITS序列分析鉴定为黄曲霉55株,寄生曲霉1株,β-微管蛋白基因序列分析结果与ITS序列分析相同;形态学鉴定为烟曲霉的37株曲霉中,ITS序列分析鉴定为37株烟曲霉复合种,β-微管蛋白基因序列分析鉴定为烟曲霉35株和仑图卢斯曲霉2株;形态学鉴定为杂色曲霉的21株曲霉中,ITS序列分析鉴定为杂色曲霉16株和未鉴定到种的曲霉5株,β-微管蛋白基因序列分析鉴定为杂色曲霉16株和聚多曲霉5株;形态学鉴定为构巢曲霉的10株曲霉中,ITS序列分析和β-微管蛋白基因序列分析均鉴定为构巢曲霉.结论 β-微管蛋白基因序列分析曲霉的分辨率较ITS序列分析高,可以将曲霉准确鉴定到种,ITS序列可以分析到曲霉复合种.Objective To study the clinical application of the ITS and β-tubulin gene regions in identification of Aspergillus spp. Methods One hundred and twenty-four Aspergillus strains that isolated from fungal rhino-sinusitis specimens were collected in Beijing Tongren Hospital, Capital Medical University from July 2007 to January 2010. They were identified by morphological and molecular methods. The first one included traditional culture, slide culture, and microscopic examination after lactophenol cotton blue stain and KOH digestion. The second one was amplifying and sequencing the part of ITS and β-tubulin gene and aligned all the sequences in the GenBank, European Molecular Biology Laboratory nucleotide sequence database, and DNA Data Bank of Japan. Results Of the 56 Aspergillus flavus identified by morphological features, fifty-five isolates were identified as Aspergillus flavus and 1 isolates was Aspergillus parasiticus by the ITS and β-tubulin gene region sequence analysis. In the 37 Aspergillus fumigatus identified by morphological method, and all the 37 isolates were identified as species complex of Aspergillus fumigatus by the ITS region sequence analysis, but through the sequence analysis of β-tubulin gene region, thirty-five isolates were identified as Aspergillus. fumigatus and 2 were Aspergillus lentulus. Twenty-one isolates were identified as Aspergillus versicolor by morphological method, but 16 of them were identified as Aspergillus. versicolor and 5 can not be identified to species level by the ITS region sequence. And by comparative-sequence analysis of β-tubulin gene region, the 5 isolates were identified as Aspergillus sydowii,the other 16 isolates were Aspergillus. versilcolor. Ten isolates were identified as Aspergillus nidulans by morphological features, the ITS and β-tubulin gene region sequence analysis. Conclusions β-tubulin gene sequencing is more suitable for identifying Aspergillus, and could identify Aspergillus spp. to species level Sequences of ITS region could only identify Asperg

关 键 词：曲霉菌属 DNA 核糖体间隔区 RNA 核糖体 微管蛋白 

分 类 号：R440[医药卫生—诊断学]