基于双抗体夹心ELISA建立人血浆肌钙蛋白I的定量测定技术  

Establishment of a technique of quantitative detection of human plasma cardiac troponin I based on double-antibody-sandwich ELISA

作　　者：孙崇云[1] 周蕾[2,3] 王晓晨[2,3,4] 赵勇[2,3] 王鑫蕊[2,3] 张平平[2,3] 杨瑞馥[2,3] 王成彬[1] 

机构地区：[1]温州医科大学检验医学院与生命科学学院,浙江温州325035 [2]军事医学科学院微生物流行病研究所、病原微生物生物安全国家重点实验室,北京100071 [3]生物应急与临床POCT北京市重点实验室(BZ0329),北京100071 [4]吉林农业大学动物科学技术学院,长春130118

出　　处：《军事医学》2016年第6期529-534,共6页Military Medical Sciences

基　　金：国家863计划资助项目(2013AA032205);北京市科技新星计划资助项目(Z151100000315086);国家科技重大专项资助项目(2013ZX10004101-003,2012ZX10004801-002-004,2012ZX10004801-004-015)

摘　　要：目的基于双抗体夹心ELISA技术建立人血浆肌钙蛋白I(cTnI)定量检测方法,便于基层医疗机构使用。方法基于抗原、抗体筛选,包被抗体、酶标抗体用量优化,建立双抗体夹心ELISA检测cTnI的方法。测定cTnI标准抗原,利用Curve Expert软件建立定量公式,根据临床大型仪器实测结果校准定量公式,实现ELISA测定D值到血浆中cTnI浓度的直接定量。结果与结论双抗体夹心ELISA测量cTnI的敏感性可达0.1ng/ml,敏感性满足临床要求。小样本临床样品的评价实验显示,双抗体夹心ELISA定量结果与临床大型仪器测得的浓度差异无统计学意义(t=0.058,P>0.05)。所建立的双抗体夹心ELISA法可用于定量检测人血浆cTnI。Objective To establish a method based on double-antibody-sandwich ELISA for quantitative detection of human cardiac troponin I（ c Tn I） in grass-roots medical institutions. Methods Double-antibody-sandwich ELISA method for detecting cardiac troponin I（ c Tn I） was established based on the screening of antigens and antibodies and the dosage optimization of coated antibody and enzyme labeled antibody. The quantitative formula was deduced by Expert Curve software according to c Tn I standard antigen detection and calibrated according to the measured results of clinical large-scale instrument. The direct quantification from D value measured by ELISA to plasma c Tn I concentration was realized. Results and Conclusion The sensitivity of double-antibody-sandwich ELISA for c Tn I detection could reach 0. 1 ng / ml,which could meet the clinical requirements. Evaluation experiment of a small amount of clinical samples displayed that no significant difference is found between the concentrations of c Tn I detected by double-antibody-sandwich ELISA and clinical large-scale instrument（ t = 0. 058,P〉 0. 05）. The double-antibody-sandwich ELISA method established in this study can be used to detect human plasma c Tn I quantitatively.

关 键 词：肌钙蛋白I 酶联免疫吸附试验 定量检测 

分 类 号：R446.6[医药卫生—诊断学]