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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。
Objective To compare the fingerprints of secreted proteins of Mycobacterium tuberculosis and non-tuberculous mycobacteria and to analyze the difference of secreted protein expression, so as to lay a foundation for the rapid identification of Mycobacterium. Methods Standard strains of Mycobacterium tuberculosis and Mycobacterium tuberculosis were selected and inactivated after culture. The secreted protein was extracted and the secreted protein was detected by surface-enhanced laser desorption / ionization time-of-flight mass spectrometry (SELDI-TOF-MS). Biomaker Wizard Software Analysis software to screen for differential proteins. Mycobacterium secretory proteins were determined repeatedly 20 times to evaluate the reproducibility of the molecular weight of mycobacteria secreted by SELDI-TOF-MS. Results AU chip can capture nearly 30 Mycobacterium secretory protein peaks, the four protein peaks for the Mycobacterium common. Compared with non-MTB protein fingerprinting, one protein peak is unique to Mycobacterium tuberculosis. SELDI-TOF-MS repeated detection of 20 mycobacterial secreted proteins showed that the relative molecular mass coefficient of variation of the same protein peak was ≤0.05%. Conclusion The discovery of differential proteins between Mycobacterium tuberculosis and non-tuberculosis mycobacteria may contribute to the identification of mycobacteria.