树鼩皮肤病原真菌的分离鉴定及基因ITS分析-中国实验动物学报.docVIP

人工饲养树鼩皮肤真菌携带的调查 尹博文，陈玲霞，李振宇，李娜，孙晓梅 （中国医学科学院/北京协和医学院医学生物学研究所树鼩种质资源中心， 云南省重大传染病疫苗研发重点实验室，昆明 650118） 【摘要】 目的 调查人工饲养条件下树鼩皮肤真菌携带情况，为实验树鼩的微生物质量控制提供依据。方法 刮取79只树鼩皮肤表皮碎屑，包括野生来源的51只和人工繁殖的F1代28只，采用沙氏青霉素瓶斜面法和沙氏平皿培养法分别进行培养，对长出的每株真菌进行取样涂片染色，显微镜镜检观察真菌形态，用酚氯仿法提取真菌DNA，PCR扩增ITS基因，测序后经BLAST进行核酸同源性比对，进一步鉴定分析。 结果 野生来源树鼩有9份样长出真菌，F1代有3份样长出真菌菌落，携带率分别15.69%和10.71%。所有真菌DNA最后进行BLAST比对鉴定后发现为7个种属的真菌，但均无致病性。结论 人工饲养的树鼩种群中皮肤真菌携带率较低，并且随繁殖代次增加而下降，未检测到皮肤病原真菌。 【关键词】 树鼩；皮肤真菌；ITS基因扩增；鉴定 【中图分类号】 Q95-33 【文献标识码】 A The Survey of Dermatophyte in Artificial Feeding of Tree Shrews YIN Bo-wen, CHEN Ling-xia, LI Zhen-yu, LI Na, SUN Xiao-mei (Center of Tree shrews Germplasm Resources, Institute of Medical Biology, Chinese Academy of Medical Sciences and Peking Union Medical College, Yunnan Key Laboratory of Vaccine Research Development on Severe Infection Diseases, Kunming 650118, China) [Abstract] Objective To investigate the pathogenic dermal fungi in tree shrews from the center of germplasm resources, to provide the basic data for the experimental tree shrews’ microbiological quality control. Methods 79 samples obtained from tree shrews were inoculated on Sabouraud’s platiculture and slant cultivation ,51 of which are from the wild origin and 28 from artificial feeding F1 generation, and they were subjected to microscope observation. Then extracted DNA with traditional methods. Amplify the ITS gene by PCR. Finally sequencing by blast of nucleotide homology analysis. Results 9 samples of the wild tree shrews turned into growing fungus,and so as 3 of the F1 generation. The carrying rate are 15.69% and 10.71% respectively. All fungi were found to be non-pathogenic followed BLAST. Conclusion The carrying rate of Dermatophyte in artificial feeding of tree shrews is very low, and which decreased with the increasing breeding generation. Pathogenic dermal fungi has not been found. [Key words] Tree Shrew ; Dermatophyte; ITS Gene Amplification; Identification  树鼩作为灵长类动物的近亲，被用于建立