分枝杆菌特征性蛋白的筛选  

Screening characteristic proteins of Mycobacterium

作　　者：胡琦[1] 杨永长[2] 黄文芳[2] 刘华[2] 喻华[2] 肖代雯[2] 黄波[1] 冯文莉[1] 

机构地区：[1]重庆医科大学医学检验系临床检验诊断学教育部重点实验室,重庆400016 [2]四川省医学科学院四川省人民医院检验科,四川成都610072

出　　处：《中国微生态学杂志》2008年第6期541-543,共3页Chinese Journal of Microecology

基　　金：四川省科技厅资助项目(2008JY0017)

摘　　要：目的研究分枝杆菌蛋白表达,筛选分枝杆菌特征性蛋白,为分枝杆菌的快速鉴定奠定基础。方法选取分枝杆菌标准菌株,提取细菌蛋白,干化学法测蛋白浓度,应用表面增强激光解吸电离飞行时间质谱技术(SELDI-TOF-MS)检测分枝杆菌蛋白表达,和非分枝杆菌蛋白指纹图谱比较,筛选分枝杆菌特征性蛋白。重复测定20次分枝杆菌蛋白标本,评价SELDI检测分枝杆菌蛋白分子量的重复性。结果耻垢分枝杆菌ATCC 14468有约20个蛋白峰,结核分枝杆菌ATCC 25177、土地分枝杆菌ATCC 15755、胞内分枝杆菌ATCC 13950、耻垢分枝杆菌ATCC 607有近40个蛋白峰。与非分枝杆菌蛋白指纹图谱相比,4个蛋白峰为分枝杆菌所特有。SELDI重复检测20次分枝杆菌蛋白标本显示同一蛋白峰的分子量变异系数≤0.05%。结论分枝杆菌有其特征性蛋白峰,可能有助于分枝杆菌的快速鉴定。Objective To screen characteristic proteins of Mycobacterium which may lay a foundation for its rapid identification. Methods Proteins of standard strains of Mycobacterium were extracted and detected by dry chemistry method. The proteins were analyzed by surface enhanced laser desorption/ionization time-of-flight mass spectra （SELDI-TOF- MS） ,compared with other bacteria, and the data was collected by Ciphergen Protein Chip software. The proteins of every stain were analyzed by SELDI for twenty times to evaluate the reproducibility of protein molecular mass. Results About twenty proteins were found in M. smegmatis 14468 ,and about forty proteins were found in M. tuberculosis ATCC 25177 ,M. terrae ATCC 15755 ,M. intrucellulare ATCC 13950 and M. smegmatis ATCC 607. Compared with other bacteria, four proteins were only found in Mycobacterium. The CV of protein in molecular weight was less than 0.05% in Mycobacterium. Conclusion The Mycobacterium has characteristic proteins,which may lay a foundation for its rapid identification.

关 键 词：分枝杆菌 特征性蛋白 表面增强激光解吸电离飞行时间质谱 

分 类 号：R372[医药卫生—病原生物学]