表面增强激光解析电离飞行时间质谱技术在结核分枝杆菌与胞内分枝杆菌差异蛋白分析中的应用

Analysis of differential proteins in mycobacterium tuberculosis and mycobacterium intracellulare with technology of surface-enhanced laser desorption/ionization time-of-flight mass spectrometry

目的对比分析结核分枝杆菌与胞内分枝杆菌菌体蛋白、分泌蛋白指纹图谱,评价表面增强激光解析电离飞行时间质谱(SELDI-TOF-MS)技术在结核病快速诊断中的价值。方法结核分枝杆菌和胞内分枝杆菌标准菌株经BACTEC960仪培养30d,灭活菌株,离心沉淀物提取菌体蛋白,上清液提取分泌蛋白。利用SELDI-TOF-MS进行蛋白质/多肽谱分析。结果结核分枝杆菌与胞内分枝杆菌菌体蛋白指纹图谱存在差异,有33个差异蛋白峰;结核分枝杆菌与胞内分枝杆菌分泌蛋白指纹图谱存在差异,有51个差异蛋白峰;结核分枝杆菌有其特征性蛋白峰。结论结核分枝杆菌与胞内分枝杆菌菌体蛋白、分泌蛋白指纹图谱有明显的差异蛋白峰,应用SELDI-TOF-MS技术可以早期、快速、简便地诊断结核病。

Objective To observe and analyze the cytosolic proteins and the secreted proteins fingerprint of mycobacterium tuberculosis and mycobacterium intracellulare,and evaluate the value of surface-enhanced laser desorption/ionization time-offlight mass spectrometry(SELDI-TOF-MS)for rapid diagnosis.Methods Mycobacterium tuberculosis and mycobacterium intracellulare standard strains were cultured in BACTEC 960 MGIT bottles for 30 days at 37 ℃,followed by bear inactivation of water bath for 30 min at 100 ℃.The cytosolic protein was extracted from the centrifugal precipitate,and the secreted protein was extracted from the supernatant.After extracting,the cytosolic proteins and secreted proteins were analyzed by SELDITOF-MS.Results 1)Thirty-three cytosolic protein peaks were different from mycobacterium tuberculosis and mycobacterium intracellulare.2)Fifty-one secreted protein peaks were different from mycobacterium tuberculosis and mycobacterium intracellulare.3)Mycobacterium tuberculosis have the characteristic protein peaks.Conclusion There are the differential protein peaks in the cytosolic proteins and the secreted proteins fingerprint of mycobacterium tuberculosis and mycobacterium intracellulare,which may provide an early,rapid and simple way for diagnosis of tuberculosis with SELDI-TOF-MS.