【摘要】 目的　研制人心肌肌钙蛋白I(cTnI)单克隆抗体 ,建立双单克隆抗体两点一步夹心酶免疫法 (简称“双抗夹心ELISA”)。方法　用亲和层析法直接从人心肌组织中提纯cTnI ,经免疫、融合、筛选、克隆等杂交瘤技术 ,制备出 3株特异性抗cTnI单克隆抗体 ,选用其中互补的 2株 2F11和9F5,分别制成生物素化抗体和酶标抗体 ,建立双抗夹心ELISA检测法。结果　用双抗夹心ELISA和美国PBM公司LifesignTroponinI检测试剂对 40名健康献血员和 3 4例急性心肌梗塞患者血中cTnI水平进行检测 ,本法最低检测限为 0 8μg/L ,两者符合率分别为 97 5%和 93 9% ,符合临床诊断要求。结论　双抗夹心ELISA特异性强 ,灵敏度、准确度和精密度较高 ,具有应用及推广价值更多还原

【Abstract】 Objectives To prepare monoclonal antibodies (McAbs) against troponin I and develop its one-step sandwich ELISA. Methods Based on purification of human cTnI by affinity chromatography, three monoclonal antibodies were obtained by hybridoma technique and the two McAbs, 2F11 and 9F5, were labeled with biotin and HRP, respectively then used to develop the one-step sandwich ELISA. Results The sensitivity of this method for cTnI was 0 8 μg/L. The cTnI in sera of 40 nomal donors and 34 patients with acute myocardial infarction (AMI) was detected by this method and the product of PBM Co USA as standard. The coincidence rates of two assays were 97 5 and 93 9, respectively, which illustrated that our one-step sandwich ELISA is suitable to clinical application. Conclusion This method is specific, simple, sensitive and accurate.更多还原