结核分枝杆菌与非结核分枝杆菌分泌蛋白指纹图谱的比较

目的比较结核分枝杆菌和非结核分枝杆菌分泌蛋白的指纹图谱,分析分泌蛋白表达的差异,为分枝杆菌的快速鉴别奠定基础。方法选取结核分枝杆菌和非结核分枝杆菌标准菌株,培养后灭活,提取分泌蛋白,用表面增强激光解吸电离飞行时间质谱(SELDI-TOF-MS)检测菌株分泌蛋白的表达,Biomaker Wizard Software分析软件筛选差异蛋白。重复测定20次分枝杆菌分泌蛋白,评价SELDI-TOF-MS检测分枝杆菌分泌蛋白相对分子质量的重复性。结果AU芯片能捕获近30个分枝杆菌分泌蛋白峰,4个蛋白峰为分枝杆菌共有。与非结核分枝杆菌分泌蛋白指纹图谱比较,有1个蛋白峰为结核分枝杆菌所特有。SELDI-TOF-MS重复检测20次分枝杆菌分泌蛋白显示,同一蛋白峰的相对分子质量变异系数≤0.05%。结论结核分枝杆菌和非结核分枝杆菌差异蛋白的发现,可能有助于分枝杆菌的鉴别。

Fingerprint Mapping of Proteins Secreted by Mycobacterium tuberculosis and Nontuberculosis Mycobacteria

Objective To compare the fingerprint maps of proteins secreted by Mycobacterium tuberculosis and by nontuber-culosis mycobacteria,analyze the difference in expressions of secreted proteins and lay a foundation of rapid differentiation of mycobacteria.Methods The standard strains of Mycobacterium tuberculosis and nontuberculosis mycobacteria were cultured and inactivated,from which the secreted proteins were extracted and determined for expression levels by SELDI-TOF-MS.The differential pro-tein was screened by Biomaker Wizard Software.The secreted proteins were determined by SELDI-TOF-MS for 20 times to evaluate the reproducibility of the method.Results Nearly 30 secreted protein peaks were captured on AU chip,of which 4 were found in all the mycobacteria.Only one differential protein in Mycobacterium tuberculosis was found by comparison of fingerprints.The same protein peak was determined by SELDI-TOF-MS for 20 times,and the result showed a variation coefficient of not more than 0.05%.Conclusion The determination of differential proteins between Mycobacterium tuberculosis and nontuberculosis mycobacteria may be helpful to differentiation of mycobacteria.