TIMP-1 ELISA检测方法条件的优化研究  

Optimization of quantitative ELISA for tissue inhibitor of metalloproteinases-1 (TIMP-1)

作　　者：邵彬[1] 聂青和[1] 王平忠[1] 苟艳子[1] 杨洁[1] 

机构地区：[1]第四军医大学唐都医院传染病科全军感染病诊疗中心,陕西西安710038

出　　处：《胃肠病学和肝病学杂志》2007年第6期601-604,共4页Chinese Journal of Gastroenterology and Hepatology

基　　金：陕西省科技攻关项目(社发攻关:2003K10G63)

摘　　要：目的进一步研究酶联免疫吸附方法(ELISA)检测人血清中基质金属蛋白酶组织抑制剂-1(TIMP-1)的最优化试验条件。同时最大程度降低成本,使血清TIMP-1 ELISA国产试剂盒价格、质量更优。方法用抗人TIMP-1单克隆抗体(mAb),辣根过氧化物酶(HRP)标记的抗TIMP-1,通过选用国产酶联板及对酶联板进行紫外线辐照处理,改变固相化条件;通过对抗体包被时间、样本温育时间等条件优化,建立简便、快速、准确的ELISA定量技术。同时复检临床408例肝病患者血清,并与优化前检测结果做以比较。结果改变固相化条件,并延长包被时间至48 h,抗原抗体反应时间为37℃2 h,可获得更佳的血清TIMP-1定量结果。结论优化后的TIMP-1测定方法在敏感性、重复性及标准曲线线性关系更好,可提高临床肝纤维化血清阳性检出率。Objective To optimize the procedure of quantitative enzyme-linked immunosorbent assay （ELISA） for tissue inhibitor of metalloproteinases-1 （TIMP-1） in human serum. Methods By using anti-TIMP-1 mAb, and anti- TIMP1 mAb with HRP, the polystyrene microtiter plates were made in China and were treated with ultraviolet radiation to improve the immobilization. The constitution of coating time and incubation time were optimized as well to establish a simple, rapid and accurate ELISA, and was used in the detection of 408 hepatitis patients. Results By improving the immobilization condition and By extending coating time to 48 h and the condition of antigen-ntibody reaction, （37 ℃ 2 h） , quantitative ELISA for TIMP-1 assay had better results. Conclusion By optimiz the sensitivity , repatition and standard curve of the quantitative ELISA for TIMP-1 is more good, and it can improve serological positive ratio of liver fibrosis.

关 键 词：基质金属蛋白酶组织抑制剂-1 酶联免疫吸附法 优化 试验条件 肝纤维化 

分 类 号：R575[医药卫生—消化系统]