云南省部分地区鸽粪中新型隐球菌的分离与鉴定  

Isolation and identification of Cryptococcus neoformans from pigeon droppings in parts of Yunnan Province

作　　者：陶星辰 尚秋菊[2] 刘晓英[1] 黄菊华[3] 杨静[2] 苏鸿雁[2] 

机构地区：[1]大理学院基础医学院,云南大理671000 [2]大理学院农学与生物科学学院 [3]大理学院公共卫生学院,云南大理671000

出　　处：《中国病原生物学杂志》2014年第10期873-876,共4页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.31060019;31100093;31460015);云南省教育厅重大专项资助项目(No.ZD2010010);云南省中青年学术带头人后备人才资助项目(No.2012HB-042);云南省高校滇西生物多样性科技创新团队基金项目(云教函〔2011〕93号);大理学院第一批专业综合改革生物科学专业综合改革建设项目(TDⅠ-05)

摘　　要：目的对云南省部分地区鸽粪中新型隐球菌进行分离鉴定,并对各个地区鸽粪中新型隐球菌带菌率进行调查。方法采集云南省4个城市鸽粪标本共473份,用黑米琼脂培养基对其进行分离,肉眼观察菌落形态、颜色,光学显微镜下观察菌体形态,对疑似菌落进行纯培养后用特异性引物CN4和CN5扩增URA5基因,阳性者鉴定为新型隐球菌,并用SPSS16.0对其结果进行统计分析。结果云南省4个地区473份鸽粪标本中共分离出152株新型隐球菌,平均阳性率为32.13%。其中大理地区阳性率为26.08%,昭通阳性率为30.05%,曲靖阳性率为81.81%,临沧阳性率为37.03%。4个地区阳性率差异有统计学意义(χ2=41.87,P<0.01)。结论云南地区鸽粪中新型隐球菌带菌率较高,且不同地区带菌率不同。Objective To isolate and identify Cryptococcus neoformansfrom pigeon droppings in parts of Yunnan Province and to investigate the rate at which those droppings tested positive for C.neoformans. Methods Sterile bamboo sticks were used to randomly collect pigeon droppings from a pigeonry.A suspension was created from 0.6g of pigeon droppings and 10 ml of sterile normal saline,and plates were coated with 100μl of black rice agar medium.The shape and color of colonies were observed and mycelial morphology was examined using an optical microscope.The plates were also used to isolate C.neoformans from pigeon droppings.The URA5 gene was amplified using the specific primers CN4 and CN5,and the serotype and mating type were verified using specific primers. Results C.neoformans was isolated from152 of 473total specimens.On average,C.neoformans was isolated at a rate of 32.13%.C.neoformans was isolated in Dali at a rate of 26.08%,in Zhaotong at a rate of 30.05%,in Qujing at a rate of 81.81%,and in Lincang at a rate of 37.03%.There were significant differences（χ^2=41.87,P〈0.01）in the rates at which C.neoformans was isolated from these 4locations. Conclusion Pigeon droppings in Yunnan tested positive for C.neoformans at a relatively high rate（32.13%）.Pigeon droppings in different areas tested positive for C.neoformans at different rates.

关 键 词：新型隐球菌 鸽粪 黑米琼脂培养基 阳性率 

分 类 号：R379.5[医药卫生—病原生物学]