【作者】 郭会时； 张寄南； 杨笛； 何农跃；

【Author】 GUO Hui-shi, ZHANG Ji-nan , YANG Di, HE Nong-yue Department of Chemistry, Shaoguan University, Shaoguan 512005, China Research Institute of Cardiovascular Disease, Hospital of Jiangsu Province, Nanjing 210029, China State Key Laboratory of Bioelectronics, Southeast University, Nanjing 210096, China

【机构】 广东省韶关学院化学系； 江苏省人民医院心血管病研究所； 东南大学生物电子学国家重点实验室；

【摘要】 文章在传统ELISA法的基础上,在进行免疫反应后,改用高效化学发光试剂AMPPD作为检测底物,并将传统两步双抗夹心法改为一步法进行免疫反应,建立了一种高灵敏度的测定人血清中心肌肌钙蛋白I(cTnI)的化学发光酶免疫分析法。采用单因素变化法和方阵滴定法优化了实验条件。该方法具有很高的灵敏度,检出限为0.02 ng·mL-1,比传统ELISA法检测限降低一个数量级以上;检测范围为0．04～36．2 ng·mL-1,比传统ELISA法扩宽了近两个数量级;方法准确可靠,重复性好,加标试验回收率介于97．5％～102．8％之间,应用于实际样品测定,结果满意。更多还原

【Abstract】 Cardiac troponin I (cTnI) is a subunit of cardiac troponin complex, which is comprosed of troponin I, troponin T and troponin C. Following the myocardial damage, the troponin complex is broken up and the individual protein components are released into the bloodstream. Cardiac troponin I has high tissue specificity because of its structural difference from the corresponding skeletal isoforms in its amino acid composition. Therefore, cTnI in the scrum of patients has been considered as a "gold standard" for diagnosis of myocardial injury. Nowadays the most extensively used method for cTnI detection is enzyme-linked immunosorbent assay (ELISA). However, it has the disadvantages of low sensitivity, and long detection period. In the present study the authors present a sensitive immunoassay for cTnI based on the concept of chemiluminescent enzyme immunoassays (CLEIA). The assay was executed as the traditional ELISA except that two major modifications were made to obtain a rapid test and a high sensitivity. The first modification is that the two-step dual monoclonal antibody "sandwich" principle of traditional ELISA was substituted by a one-step immunoreaction mode, which decreased the turnaround time greatly. The second modification is that a chemiluminescent substrate (AMPPD) and chemiluminescencc detection techniques were used to replace the colori-metric substrate and colorimetric detection methods, which elevated the sensitivity remarkably. The detection procedure of the assay could be fulfilled within 75 min. A linear range between the luminescent signal current and the concentration of cTnI from 0. 04 to 36. 2 ng · mL-1 and a detection limit of 0. 02 ng · mL-1 were obtained. The established method was tested by determining cTnI in real samples using ELISA for comparison analysis, and good results were obtained.更多还原