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目的分析对比结核分枝杆菌(MTB)与非结核分枝杆菌(NTM)分泌蛋白指纹图谱,寻找检测MTB的蛋白标志物。方法经BACTEC 960仪培养出结核分枝杆菌标准菌株,提取分泌蛋白,应用表面增强激光解析电离飞行时间质谱(SELDI-TOF-MS)检测MTB蛋白表达,和NTM标准株、临床MTB及其他肺部炎症细菌蛋白指纹图谱进行对比分析。结果分枝杆菌分泌蛋白共检出83个蛋白峰,3145、3895m/z在所有分枝杆菌均高表达,其他肺部炎症分离株未见表达。NTM与MTB标准株比较,有51个蛋白峰分泌蛋白存在差异,MTB有5个特征性蛋白峰(2930、3878、4103、4902、5804m/z)。结论分枝杆菌有其特征性分泌蛋白,有助于分枝杆菌快速鉴定。应用SELDI-TOF-MS技术对BACTEC 960阳性培养液的上清液蛋白组成进行分析,有助于MTB的快速鉴别。
Objective To compare and analyze the fingerprints of the secreted proteins of Mycobacterium tuberculosis (MTB) and non-tuberculous mycobacteria (NTM) in order to find out the protein markers for detecting MTB. Methods Standard strains of Mycobacterium tuberculosis were cultured by BACTEC 960 instrument. The secretory proteins were extracted and MTB protein expression was detected by surface-enhanced laser desorption / ionization time-of-flight mass spectrometry (SELDI-TOF-MS). NTM standard strains, clinical MTB and other lung Inflammatory bacterial protein fingerprinting comparison analysis. Results Mycobacterium secretory proteins detected a total of 83 protein peaks, 3145,3895m / z were highly expressed in all mycobacteria, other lung inflammatory isolates were not expressed. There were 51 protein peak secretory proteins in NTM compared with MTB standard strains, and MTB had 5 characteristic protein peaks (2930, 3878, 4103, 4902, 5804 m / z). Conclusion Mycobacterium has its characteristic secreted protein, which is helpful for rapid identification of mycobacteria. SELDI-TOF-MS technology was used to analyze the protein composition of the supernatant of BACTEC 960 positive culture medium, which is helpful for the rapid identification of MTB.