应用蛋白质组学技术建立Behcet病相关葡萄膜炎诊断模型并筛选血清学标记物

发布时间：2018-05-02 08:35

本文选题：Behcet病相关葡萄膜炎 + 蛋白组学　；参考：《北京协和医学院》2010年博士论文

【摘要】：目的：应用蛋白质组学技术建立Behcet病相关葡萄膜炎诊断模型并筛选血清学标记物。方法：152例血清样本包括Behcet病相关葡萄膜炎活动期、缓解期、小柳原田病(VKH)各31例及59例正常对照,所有样本同批进行MALDI-TOF-MS结合蛋白质芯片分析并绘制蛋白指纹图谱；应用BPS软件建立诊断模型并进行独立样本验证；应用磁珠结合、电泳及质谱技术分离、纯化和鉴定差异蛋白；应用ELISA和免疫沉淀技术验证所得蛋白质。结果：1)在质荷比1000-50000m/z范围内将Behcet病相关葡萄膜炎同VKH及正常对照相比较,建立Behcet病相关葡萄膜炎血清蛋白质组学诊断模型,经独立验证敏感性为84.62%,特异性为90.91%。2)6个蛋白峰(M1711.82、M8688.13、M11490.3、 M11548.2、M39478.0和M58518.8)在Behcet病相关葡萄膜炎活动期与缓解期患者中有表达差异(p0.05)；其中M1711.82和M8688.13在活动期表达低于缓解期,M11490.3、M11548.2、M39478.0和M58518.8高于缓解期；经分离、纯化、鉴定后并通过ELISA和免疫沉淀验证,M11490.3和M11548.2为血清淀粉样蛋白A (Serum Amyloid A, SAA),M39478.0为载脂蛋白Al(Apolipoprotein Al, ApoAl)。结论：应用蛋白质组学技术建立了Behcet病相关葡萄膜炎血清蛋白质组学诊断模型,并筛选出血清淀粉样蛋白A和载脂蛋白A1作为Behcet病相关葡萄膜炎活动期的血清学标记物。
[Abstract]:Objective: to establish a diagnostic model of Behcet's disease associated uveitis by proteomics and to screen serological markers. Methods Behcet's disease associated uveitis in active stage, remission stage, willow Harada disease in 31 cases and normal control in 59 cases were collected from 152 serum samples. All samples were analyzed by MALDI-TOF-MS binding protein chip and protein fingerprints were plotted. BPS software was used to establish diagnostic model and independent sample validation; magnetic bead binding electrophoresis and mass spectrometry were used to isolate and identify differential proteins; ELISA and immunoprecipitation techniques were used to verify the proteins. Results: the serum proteomics diagnosis model of Behcet disease associated uveitis was established by comparing Behcet's disease associated uveitis with VKH and normal controls within the range of quality-charge ratio (1000-50000m/z). The sensitivity and specificity of M1711.82, M11548.13, M11490.3, M11548.2, M39478.0 and M58518.8) were significantly higher in patients with active and remission uveitis associated with Behcet's disease than in patients with remission, and the expression of M1711.82 and M8688.13 in active phase was lower than that in remission stage, and the expression of M1711.82 and M8688.13 in active phase was lower than that in remission phase, and the expression of M1711.82 and M8688.13 was significantly higher than that in remission stage; after separation, the expression of M1711.82 and M8688.13 in active phase was lower than that in remission stage. After purification, identification and identification by ELISA and immunoprecipitation, M11490.3 and M11548.2 were identified as serum amyloid A serum Amyloid A, SAA Agna M39478.0 as apolipoprotein Al(Apolipoprotein Al. Conclusion: the serum proteomics diagnosis model of Behcet's disease associated uveitis was established by using proteomics technique, and serum amyloid A and apolipoprotein A1 were screened as serological markers in the active phase of Behcet's disease associated uveitis.
【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2010
【分类号】：R773

【共引文献】