微孔板化学发光免疫分析法检测人血清中三碘甲状腺原氨酸

Microplate chemiluminescence immunoassay for determination of triiodothyronine in human serum

建立了一种定量人血清中三碘甲状腺原氨酸(T3)的化学发光免疫分析方法,使用辣根过氧化物酶(HRP)标记T3抗原,HRP催化鲁米诺-过氧化氢化学发光为检测体系.对几种影响因素,如包被抗体、酶标物的滴度进行选择,对温育及检测时间等进行优化.进行了方法学评价:灵敏度为0.07ng/mL;批内和批间变异均小于10%;回收率在80%～90%之间;与结构相似物T4及rT3均无明显交叉.使用本方法与现有的发光试剂盒进行比对,相关性良好,相关系数为0.9547.

A microplate chemiluminescence immunoassay（CLIA） was developed for the clinical determination of Triiodothyronine（T3） in human serum,and the luminal-hydrogen peroxide was used as chemiluminescence system catalyzed by horseradish peroxidase（HRP）.Several physicochemical parameters such as the dilution ratios of coated antibody and HRP-T3 conjugate was chose,the incubation time and detection time were optimized.Methodology evaluation was done：the detection limit was 0.07 ng/mL;intra-assay and inter-assay coefficient of variation（CV） was less than 10%;the recovery was 80%-90%;there was not have obviously cross-reactivity of T4 and rT3 with the similar structure.Compared with a commercial CLIA kit,there was a good correlation between the two methods,the correlation coefficient was 0.9547.