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目的初步探讨结核病患者与正常人之间血清蛋白质的差异,以期筛选出新的结核病实验诊断标志物。方法对10例菌阳肺结核(阳性组)、10例菌阴肺结核(阴性组)、10例肺外结核(肺外组)及10例正常人(正常组)分组混合血清应用17cmpH4～7IPG胶条进行双向凝胶电泳(two-dimensional electrophoresis,2-DE),每组平行电泳3块胶条。电泳结束后对2-DE胶条进行考马斯亮蓝染色,染色后用Umax powerlook2000扫描仪以投射方式、300dpi分辨率扫描获取2-DE图谱,然后用2-DE分析软件Image Master2D Platium5.0进行分析。结果在正常组、阳性组、阴性组、肺外组中,分别平均可以检测到310±64、307±33、296±54、267±23个清晰可见的蛋白质点。正常组与阳性组、阴性组、肺外组的匹配率分别为(74±2)%、(76±2)%和(68±5)%。通过对4组2-DE图谱比较,阳性组与正常组间有21个差异蛋白质点(其中9个上调),阴性组与正常组间有18个差异蛋白质点(其中8个上调),肺外组与正常组间有24个差异蛋白质点(其中7个上调)。将阳性组、阴性组、肺外组合并为结核组与正常组比较有3个差异蛋白质点(其中2个上调)。结论结核病患者和正常人血清蛋白质存在差异,有望从血清中筛选出新的结核病实验诊断标志物。本研究建立了两者间血清蛋白质组差异蛋白质的初步模型,但有待进一步验证并进行差异蛋白质鉴定。
Objective To investigate the differences of serum proteins between patients with tuberculosis and normal subjects so as to screen out new experimental diagnostic markers of tuberculosis. Methods 10cases of positive pulmonary tuberculosis (positive group), 10 cases of negative pulmonary tuberculosis (negative group), 10 cases of extrapulmonary tuberculosis (pulmonary extranodal group) and 10 normals (normal group) mixed serum 17cmpH4 ~ 7IPG strips Two-dimensional gel electrophoresis (2-DE) was performed, and each strip was electrophoresed in 3 strips. After electrophoresis, the 2-DE strips were stained with Coomassie brilliant blue. After staining, 2-DE patterns were obtained by scanning with a Umax powerlook2000 scanner and scanning with a resolution of 300 dpi, and then analyzed with 2-DE analysis software Image Master2D Platium5.0 . Results In the normal group, the positive group, the negative group and the extrapulmonary group, an average of 310 ± 64,307 ± 33,296 ± 54,267 ± 23 clearly visible protein spots were detected respectively. The matching rates of normal group, positive group, negative group and extrapulmonary group were (74 ± 2)%, (76 ± 2)% and (68 ± 5)%, respectively. By comparing 4-group 2-DE maps, there were 21 differential protein spots (9 of which were up-regulated) between the positive group and the normal group, 18 differential protein spots (8 of which were up-regulated) between the negative group and the normal group, There were 24 differential protein spots (7 of which were up-regulated) between the group and the normal group. The positive group, negative group, extrapulmonary combination and tuberculosis group compared with the normal group there are three different protein spots (two of which are upregulated). Conclusion There are differences in serum proteins between tuberculosis patients and normal people, and it is expected to screen out new experimental diagnostic markers of tuberculosis from serum. This study established a preliminary model of the differential protein between the two serum proteomes, but needs further verification and differential protein identification.