临床少见丝状真菌的实验室诊断  

Laboratory diagnosis of rarely filamentous fungi

作　　者：尹秀云[1] 梁钰英 于农[1] 曾利军[1] 李一鹤[1] 蒋虔[1] 徐建民[1] 杨哲[1] 李好莲 陈建魁[1] 陈水平[1] 

机构地区：[1]军事医学科学院附属医院检验科,北京100071

出　　处：《军事医学》2015年第11期855-858,共4页Military Medical Sciences

基　　金：国家科技重大专项课题资助项目(2013ZX10004203);国家重大科学仪器设备开发专项资助项目(2012YQ18011708);军事医学科学院附属医院创新科研基金课题资助项目(ZH-2014-09)

摘　　要：目的通过对临床少见丝状真菌的分离和分析鉴定,探讨尖端赛多孢子菌、裂褶菌、短柄帚霉、匍枝根霉、茄病镰刀菌等临床不常见丝状真菌的实验诊断方法,为临床明确诊断真菌感染种类提供科学依据。方法从临床标本培养出的5株真菌用形态学方法未能作出准确鉴定,通过聚合酶链反应(PCR)和基因测序方法分析核糖体转录间隔区域ITS和ITS2序列,在NCBI网站应用Blastn对获得的基因序列进行同源性分析及利用Gen Bank中的系统发育软件自动生成系统发育树,以确定丝状真菌的种。结果 r DNA-ITS序列分析结合形态学分析,5株菌均可鉴定到种的水平,鉴定结果分别是:尖端赛多孢子菌、裂褶菌、短柄帚霉、匍枝根霉和茄病镰刀菌。结论传统形态学方法,极易受检验人员经验水平影响,对于罕见菌株甚至不能作出鉴定,利用真菌的ITS序列的特异性,可快速、准确将大部分丝状真菌鉴定到种水平。Objective To investigate the laboratory diagnosis of filamentous fungi which are rarely seen in clinical practice.Methods Five strains of fungi were isolated from clinical samples and initially identified by the morphological method.Total DNA of fungi was extracted and amplified by the PCR method using universal primers of ITS2-ITS4 gene, respectively.The PCR products were sequenced and the obtained sequences were then analyzed by the blastn program incorporated in NCBI.Results The five strains of fungi were diagnosed as Scedosporium apiospermum,Schizophyllum commune,Scopulariopsis brevicaulis,Rhizopus stolonifer,and Fusarium solani.Conclusion The laboratory diagnosis of filamentous fungi which rarely occur in clinical practice should integrate various methods,including morphological, microbiological,and molecular biological methods.

关 键 词：RDNA 丝状真菌 内转录间隔区 形态学鉴定 序列分析 

分 类 号：R440[医药卫生—诊断学]