某校园环境中烟曲霉菌的分离及CSP分型鉴定  

Isolation and genotype identification of Aspergillus fumigatus from different environments of a campus

作　　者：吴金燕[1,2] 张鹏程[1] 易国辉[1] 何小稳[1] 郭虹[1] WU jinyan ZHANG pengcheng YI guohui HE xiaowen GUO hong(Public Laboratory of Hainan Medical University, Haikou , Hainan 571199, China Laboratory for Conversation and Utilization of Bio-Resources, Yunnan University, Kunmin, Yunnan 650091, China)

机构地区：[1]海南医学院科学实验中心,海南海口571199 [2]云南大学云南省生物资源保护与利用重点实验室,云南昆明650091

出　　处：《中国热带医学》2017年第7期655-658,共4页China Tropical Medicine

基　　金：海南省自然基金(No.20158374);海南省教育厅基金(No.Hjkj2012-36)

摘　　要：目的通过采集某校园环境标本,分离、鉴定出烟曲霉菌并对其进行分型研究,了解该校园环境中烟曲霉菌基因型及其分布情况。方法釆集该校内土壤、腐叶来源样本进行培养,10%KOH染色及乳酸棉兰染色,显微镜下观察;提取基因组DNA并扩增β-微管蛋白(β-tubulin)基因以鉴定烟曲霉菌,利用烟曲霉细胞表面蛋白(CSP)对其进行分型。结果从土壤、腐叶各20份共40份样本中共分离到43株真菌,其中土壤样本中分离到19株,腐叶样本中分离到24株;经β-tubulin基因PCR扩增鉴定,22株为烟曲霉菌,包括土壤来源的8株和腐叶来源的14株;所获的烟曲霉菌经CSP基因扩增及序列测定,分为2个基因型:t01型和t25型,其中t25型为新增CSP基因型。结论烟曲霉菌在环境中普遍存在,不同地区其烟曲霉菌种类存在差异,该研究中所发现的烟曲霉菌CSP基因型t25为新增型。Objective To understand the distribution and genotypes of Aspergillus fumigatus isolated from soil and rottingleaves in the campus of a university.Methods The samples were collected from soil and rotting leaves in the campus ofauniversity in Hainan Province and cultured to isolate Aspergillus fumigatus.Aspergillus fumigatus were observed under amicroscope after coloring by 10% KOH and lactic acid Medan.The genomic DNA of Aspergillus fumigatus was extracted and β-tubulin gene was amplified to confirm Aspergillus fumigatus.Aspergillus fumigatus were typed by the cell surface protein（CSP）amplification.Results Totally 43 fungi were isolated from 40 samples including 20 soil samples and 20 rot leaf samples,among which 19 strains were isolated from soil samples and 24 strains were isolated from rot leaf samples.Twenty-two strainsincluding 8 from soil and 14 from rot leaves were identified as Aspergillus fumigatus by PCR amplification of β-tubulin gene,and the 22 Aspergillus fumigatus were divided into two genotypes（t01 and t25） by amplifying and sequencing CSP.Conclusion Aspergillus fumigatusis widespread in the environment and the species are relatively simple in the campus.TheCSP type t25 was newly reported.

关 键 词：烟曲霉菌 Β-微管蛋白 细胞表面蛋白 基因型 

分 类 号：R379.6[医药卫生—病原生物学]