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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 isolation and identification of clinical filamentous fungi, and to explore the methods of laboratory diagnosis of filamentous fungi, such as Acinetobacter spp. Schizophylla, Schizophylla, Rhizopus stolonifer and Fusarium solani , To provide a scientific basis for the clinical diagnosis of fungal infections. METHODS Five strains of fungi cultured from clinical specimens were not identified by morphological methods. ITS and ITS2 sequences were analyzed by polymerase chain reaction (PCR) and gene sequencing methods. Blastn was used on the NCBI website to obtain Homology analysis of the gene sequence and phylogenetic tree was automatically generated using phylogenetic software in Gen Bank to identify species of filamentous fungi. Results The results of r DNA-ITS sequence analysis and morphological analysis showed that all the five strains could be identified as species of Acinetobacter ssp., Schizophylluis marcescens, Rhizoctonia solani, Fusarium. Conclusion The traditional morphological methods are highly susceptible to the experience level of the inspectors. Even rare strains can not be identified. Utilizing the ITS sequence specificity of fungi, most of the filamentous fungi can be quickly and accurately identified.