人可溶性B7-H3酶联试剂盒的研制及在肝病患者血清中水平的检测  

Establishment of a sandwich ELISA for testing serum sB7-H3 and detection of sB7-H3 levels in liver disease

作　　者：李淑湘[1] 张光波[1] 孙海洪[2] 高鑫[1] 张学光[1] 

机构地区：[1]苏州大学医学生物技术研究所,江苏苏州215007 [2]苏州大学医学部生物化学与分子生物系,江苏苏州215123

出　　处：《细胞与分子免疫学杂志》2012年第1期84-86,90,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自科学基金资助项目(30901789);江苏省卫生厅科技项目(30307001494)

摘　　要：目的:建立定量检测人的可溶性B7-H3(sB7-H3)酶联检测试剂盒,并分析肝病患者外周血中sB7-H3水平及其临床意义。方法:在已获得2株识别位点不同的小鼠抗人B7-H3单克隆抗体(4H7和2E6)基础上,采用单抗4H7为包被抗体,以经生物素标记的抗体2E6为检测抗体,以重组人可溶性B7-H3为标准品,建立可溶性B7-H3分子的酶标检测方法。在此基础上对健康献血者和不同肝病患者的血清样本进行了检测并分析其临床意义。结果:研制了sB7-H3酶联检测试剂盒,检测的线性范围是8.192～2 000 ng/L,批内、批间变异系数分别为3.35%和6.97%。酶标检测板在4℃放置1个月,变异系数(CV%)<±8.6,回收率为94%～117%,特异性试验显示无交叉反应,提示该检测试剂盒具有良好的灵敏度、稳定性和特异性。用该试剂盒分析不同肝病患者外周血sB7-H3水平后发现,(1)乙肝患者以及乙肝合并肝硬化或肝癌患者与健康对照组之间并不存在差异(P>0.05);(2)重症肝炎患者血清B7-H3水平低于健康对照组(P=0.0183);(3)血吸虫性肝硬化患者血清B7-H3水平显著高于健康对照组(P<0.01)。结论:建立了灵敏、特异的人sB7-H3酶联检测试剂盒,对肝病患者检测的结果表明,不同的肝病患者外周血sB7-H3表达水平不同,显示该试剂盒具有潜在的应用价值。AIM： To establish quantitative ELISA for soluble sB？-H3 and evaluate its clinical application. METHODS： Two mAbs of mouse anti-human sBT-H3 （4H7 and 2E6）established by our lab were used. The mAb 4H7 was used as coating antibody and the 2E6 as a sandwich antibody, which recognized a different epitope and was labeled by biotin. The serum sBT-H3 levels of healthy volunteers and patients were measured by this method. RESULTS： Sandwich ELISA was established with the linear range covering from 8. 192 ng/L to 2 000 ng/L. The outcomes of repeatability test, recovery test, stability test and specificity test were quite good. The increased levels of sB7-H3 were found in schistosomiasis cirrhosis, the reduced levels in severe hepatitis and the difference in both situations showed significance. CONCLUSION： A specific and sensitive sandwich ELISA is developed, the detection in liver disease indicates that the ELISA kit provides a potential tool in the clinical application.

关 键 词：可溶性B7-H3 ELISA 肝病 

分 类 号：R392.33[医药卫生—免疫学]