应用TaqMan MGB探针实时荧光定量PCR结合分离培养监测分析灰仓鼠细菌多样性  

Monitoring and analysis on bacterial diversity of Cricetulus migratorius by TaqMan MGB probe real- time fluorescence quantitative PCR and bacteria isolation culture

作　　者：高正琴[1] 邢进[1] 冯育芳[1] 关伟鸿[1] 贺争鸣[1] 岳秉飞[1] 

机构地区：[1]中国食品药品检定研究院,北京100050

出　　处：《药物分析杂志》2014年第2期221-227,共7页Chinese Journal of Pharmaceutical Analysis

基　　金：国家科技支撑计划(重大疾病动物模型和实验动物资源的标准化及评价体系的建立)(2011BAI15B01)

摘　　要：目的:监测分析中国灰仓鼠细菌的多样性,收集更多的质量信息,获得更全面的微生物背景知识,为灰仓鼠的种群建立、生物净化和动物模型的建立提供科学的数据依据。方法:60只成年灰仓鼠来自新疆地区,安乐死后剖解采集标本。应用TaqMan MGB探针实时荧光定量PCR和常规微生物学方法进行细菌多样性研究。结果:本研究采用常规微生物学方法从灰仓鼠中鉴定出179株细菌,包括大肠埃希菌、鲍氏志贺菌、肺炎克雷伯菌、阴沟肠杆菌、栖冷克吕沃尔菌、嗜肺巴斯德菌、溶血性巴斯德菌、产气巴斯德菌、铜绿假单胞菌、栖稻假单胞菌、门多萨假单胞菌、粪产碱杆菌、阴道加特纳菌、卡它莫拉菌、莫拉克斯氏菌、CDC group EF-4a、特氏纤维单胞菌、戊糖片球菌、金黄色葡萄球菌、溶血性葡萄球菌、腐生葡萄球菌、模仿葡萄球菌、头状葡萄球菌、孔氏葡萄球菌、木糖葡萄球菌、鸡葡萄球菌、里昂葡萄球菌、山羊葡萄球菌、粘滑罗斯菌、麻疹孪生球菌、肺炎链球菌、咽峡炎链球菌、血链球菌、牛链球菌Ⅰ型、乳房链球菌、少酸链球菌、无乳链球菌、粪肠球菌、黄肠球菌、浅绿气球菌、解酪蛋白巨大球菌、拥挤棒杆菌、人皮肤棒状杆菌、蜂房芽孢杆菌、短芽孢杆菌、环状芽胞杆菌。分离培养鉴定出12个科21个属48个种179株细菌,肠杆菌科是主要优势菌群,葡萄球菌科、肠球菌科、假单胞菌科、巴斯德菌科、链球菌科是次优势菌群。应用TaqMan MGB探针实时荧光定量PCR方法从灰仓鼠中检出167份核酸阳性标本,包括支原体、泰泽氏菌、空肠弯曲菌、CAR菌、幽门螺杆菌、肝螺杆菌、嗜肺巴斯德菌。TaqMan MGB探针实时荧光定量PCR检测结果显示支原体是最主要的优势菌群,泰泽氏菌、空肠弯曲菌、CAR菌、幽门螺杆菌、肝螺杆菌是次优势菌群。结论:TaqMan MGB探针实时荧光定量PCR结�Objective： To monitor and analyze the bacteria diversity of Cricetulus migratorius in China,gather more information about the quality of Cricetulus migratorius,and gain more comprehensive knowledge of their microorganism background so as to provide a scientific data basis for population building and biological cleaning and animal model establishment. Methods： A total of 60 adult Cricetulus migratorius brought from Xinjiang area were dissected after being euthanized and specimens were collected.Bacterial diversity research was carried out by TaqMan MGB probe real-time fluorescence quantitative PCR and routine microbiological methods. Results： A total of 179 bacterial isolates were characterized from Cricetulus migratorius by the routine microbiological method,including Escherichia coli,Shigella boydii,Klebsiella pneumoniae,Enterococcus cloacae,Enterobacter aerogenes,Kluyver cryocrescens,Pasteurella pneumotropica, Mannheimia haemolytica,Pasteurella aerogenes,Pseudomonas aeruginosa,Pseudomonas oryzihabitans,Pseudomonas mendocina,Alcaligenes faecalis,Gardnerella vaginalis,Moraxella catarrhalis,Moraxella and CDC group EF-4a,Cellulomonas turbata,Pediococcus pentosaceus,Staphylococcus aureus,Staphylococcus haemolyticus,Staphylococcus saprophyticus,Staphylococcus simulans,Staphylococcus capitis,Staphylococcus cohnii,Staphylococcus xylosus,Staphylococcus gallinarum,Staphylococcus lugdunensis,Staphylococcus caprae,Rothia mucilaginosa,Gemella morbillorum,Staphylococcus pneumoniae,Streptococcus anginosus,Globicatella sanguinis,Streptococcus bovis I,Streptococcus uberis,Streptococcus acidominimus,Streptococcus agalactiae,Enterococcu faecium,Enterococcus casseliflavus,Aerococcus viridans,Macrococcus caseolyticus,Leifsonia aquatica,Dermabacter hominis,Bacillus alvei,Brevibacillus brevis and Bacillus circulans in 96 isolates.By the routine microbiological method,12 families,21 genera and 48 species of 179 strains were isolated and identified.The major dominant bacteria group was Eenterobacteriaceae,followed by Staphylococc

关 键 词：灰仓鼠 细菌多样性 TAQMAN MGB探针 实时荧光定量PCR 常规微生学方法 分离培养 生化鉴定 监测分析 

分 类 号：R917[医药卫生—药物分析学]