血清肌钙蛋白-I均相化学发光免疫检测方法的建立

Establishment of homogeneous chemiluminescent immunoassy for detecting serum cardiac troponin I

目的：采用光激化学发光免疫分析技术建立血清肌钙蛋白-I（cTnI）均相免疫检测方法。方法：使用兔抗人cTnI多克隆抗体包被受体微球,羊抗人cTnI单克隆抗体进行生物素化与链霉亲和素包被的供体微球共同构成检测试剂,优化反应条件并进行方法学评价。结果：本方法快速、敏感,检测时间为17.5 min;分析灵敏度为0.045 ng/mL,功能灵敏度为0.053 ng/mL;回收率为104.96%～108.21%;批内精密度为3.88%～5.53%,批间精密度为7.60%～8.75%;特异性分析中各种内源性物质的干扰率均〈10%;本方法测得的cTnI正常参考值上限为1.05 ng/mL。与直接化学发光检测法具有良好的相关性（r2=0.979）。结论：本研究建立的方法能够用于定量检测血清cTnI,该方法具有均相、免清洗分离等优点,为临床提供了一个便捷高敏的检测平台。

Objective To establish homogeneous immunoassay for detecting serum cardiac troponin I（cTnI） by using light induced chemiluminescent immunoassay（LiCA）. Methods Polyclonal antibodies of cTnI were coated on the receptor particles, monoclonal antibodies of cTnI were biotinylated, and the donor particles were coated with streptavidin, all of which were composed of LiCA reagents. The optimal test conditions and analytical performance of the detection method were studied. Results The method was rapid, sensitive, and detection time was 17.5 min.The analytical sensitivity was 0.045 ng / mL and the functional sensitivity was 0.053 ng / mL.The recovery rate was 104.96%-108.21%; The within-run and the between-run coefficients of variation were 3.88%- 5.53% and 7.60%-8.75%, respectively. The interference rates for the endogenous substances were less than 10%. The reference value of cTnI was less than 1.05 ng / mL;Results of cTnI LiCA correlated well with direct chemiluminescence detection（r2= 0.979）. Conclusions This approach can be used for the quantitative detection of serum cTnI, and it is homogeneous and is free of clean separation. It provides a convenient, highly sensitive detection platform for clinical practice.