【摘要】 目的测定孢子丝菌的基因序列并同参考序列进行系统发育学分析,鉴定孢子丝菌种,探讨准确高效的鉴别菌种的方法。方法提取既往根据组织病理鉴定为申克孢子丝菌的16株临床孢子丝菌株的基因组DNA,利用核糖体内部转录间隔区(Ribosomal Internal Transcribed Spacer,ITS)和部分β微管蛋白(β-tubulin)序列进行PCR扩增,扩增序列同来源于基因库(GenBank)的参考序列进行系统发育学分析。结果基因组DNA分别扩增出约600bp的ITS及约400bp的β-tubulin编码基因,测序后构建的系统发育树分析显示所有的菌株均位于球形孢子丝菌分支上,该16株菌并非申克孢子丝菌(Sporothrix schenckii,S.schenckii),而是球形孢子丝菌(Sporothrix globosa,S.globosa)。结论中国东北孢子丝菌病致病菌可能为球形孢子丝菌或并非只是单一的申克孢子丝菌,分子生物学鉴定方法更为准确,为临床快速有效鉴别孢子丝菌菌种奠定基础。更多还原

【Abstract】 In this study,we conducted the phylogenetic analysis between the Sporothrixgene sequences and the reference sequences by elucidating the molecular characteristic,investigating more accurate and efficient identification of the pathogen of the disease.The isolates from patients which had been identified as Sporothrix schenckii was analyzed by molecular biology method.The phylogeny were inferred by DNA sequence analyses based on datasets of the ribosomal internal transcribed spacer(ITS)and a combined ITS and partialβ-tubulin region.The sequences with reference sequences which were retrieved from GenBank were used for phylogenetic analyses.Results showed that approximate 600 bp ITS gene and 400bpβ-tubulin gene were synthesized,and all of the isolates were clustered in a distinct clade with a type of S.globosa.Therefore,the 16 isolates were not S.schenckii,but S.globosa.It’s indicated that S.schenckiiis not the only causative agent of sporotrichosis,or even S.globosa may be the major cause of sporotrichosis in China.Identification of pathogens of sporotrichosis methods is more accurate than traditional methods from the perspective of molecular biology,which would lay the foundation for identification of clinical Sporothrixspecies.更多还原