血清HER2蛋白化学发光免疫检测试剂的开发及分析性能评估

Development and performance evaluation of chemiluminescent immunoassay reagent for serum HER2 protein

目的开发一种自动化、经济、定量、操作简便、样本易获得的人表皮生长因子受体2(HER2)的检测方法。方法本研究采用体外诊断化学发光免疫分析法定量检测血清中HER2蛋白的含量,将HER2单克隆抗体作为一抗包被于羧基磁珠上,并将另一二抗标记于吖啶酯上,进行双抗体夹心检测。结果本研究建立的HER2蛋白检测方法的检测范围为0~500 ng/ml,精密度为CV<5%,分析灵敏度(LOD)为0.075 ng/ml,功能灵敏度(LOQ)为0.093 ng/ml,线性范围为2.135~780.427 ng/ml,用HAMA阳性血清测试抗干扰实验,显示对本试剂盒无干扰,加速稳定性实验显示试剂可稳定存放1年。结论与在售化学发光试剂相比,本文建立的HER2蛋白检测方法在各项分析性能表现优异。成功搭建了HER2蛋白检测的原型试剂。

Objective To develop an automated,economical,quantitative,easy to operate and easily obtained sample detection of human epidermal growth factor receptor-2（HER2）. Methods Chemiluminescence immunoassay was used in vitro to quantitatively detect the content of HER2 protein in serum. One HER2 monoclonal antibody（primary antibody） was coupled to the carboxyl magnetic beads,and another HER2 monoclonal antibody（secondary antibody） was labeled to acridinium ester,to assemble a double antibody sandwich method detection reagent. Results The detection range of HER2 protein in this research was from 0 to 500 ng/ml,and precision was CV lower than 5%. The limited of detection and quantification were 0. 075 and 0. 093 ng/ml,respectively. Linearity range is from 2. 135 to 780. 427 ng/ml. The antiinterference test of HAMA positive serum test showed no interference to this kit. The accelerated stability experiment showed that the reagents could be stably stored for 1 years. Conclusions Compared with Her2 protein chemiluminescence reagents sold on the market,the detection in this study has an excellent expression in various analytical performance,which establish a HER2 protein chemiluminescence detection prototype successfully.