半巢式PCR扩增联合基因测序鉴定丝状真菌的实验研究  

Experiment Study of Using Semi-nested PCR Amplification Combined Gene Sequencing to Identify Filamentous Fungi

作　　者：刘先宁 胡淑玲[2] 安娜[1] 王亚妮[1] 朱娟莉[1] 朱秀萍[1] 

机构地区：[1]西安市第一医院陕西省眼科研究所陕西省眼科学重点实验室,西安710002 [2]陕西省人民医院检验科,西安710068

出　　处：《现代检验医学杂志》2013年第5期48-49,52,共3页Journal of Modern Laboratory Medicine

基　　金：西安市科技局项目（SF 09023）.

摘　　要：目的 采用分子生物学方法鉴定痰标本培养分离出的丝状真菌菌种.方法 收集陕西省人民医院检验科2012年2月～5月痰标本分离的丝状真菌菌落20份,采用半巢式PCR联合基因测序法及形态学方法同时鉴定菌种.结果 基因测序法：烟曲霉菌13株,黑曲霉菌3株,土曲霉菌1株,绿色木霉1株,娄地青霉菌1株,淡紫拟青霉菌1株.形态学方法：17株曲霉菌与基因序列分析法结果一致,其余3例未能鉴别出菌种.结论 半巢式PCR扩增联合基因测序鉴定丝状真菌快速、简便、准确,可克服传统表型法鉴定真菌的缺点.Objective To identify filamentous fungus which were isolated and cultured from sputum specimens with molecular biology method. Methods Collected 20 copies of filamentous fungi positive colonies isolated in clinical laboratory of Shaanxi Provincial People＇s Hospital from February to May 2012. Then identified pathogens using PCR combined gene sequencing and morphological method. Results Gene sequencing：The 20 copies of filamentous fungus identification results were 13 cop-ies aspergillus fumigatus, 3 copies aspergillus niger, 1 copy soil aspergillus, 1 copy triehoderma viride, 1 copy penicillium ro-queforti and 1 copy penicillium paecilomyces. Morphological method：The result of 17 copies aspergillus were same with the gene sequencing,and the remaining three copies were failed to identify the species. Conclusion Using Semi-nested PCR am- plification combined gene sequencing to identify pathogens is fast,simple and accurate and can overcome the shortcoming of traditional phenotype method.

关 键 词：痰标本 丝状真菌 半巢式PCR联合基因测序 

分 类 号：Q503[生物学—生物化学] R379.2[医药卫生—病原生物学]