磁微粒化学发光法测定人尿液/血浆中儿茶酚胺类物质

Determination of catecholamines in human urine/plasma by magnetic particle chemiluminescence method

目的建立血浆和尿液中游离儿茶酚胺类物质的化学发光免疫检测方法。方法将血浆或尿液标本进行提取、酰化、释放、甲基化处理,处理后的待测物分别与固相化的特异性抗体反应,再加入亲和素标记的辣根过氧化物酶(HRP),与待测抗原上的生物素结合,可以形成固相化的抗体-抗原-HRP免疫反应复合物,HRP催化底物发光。结果建立的肾上腺素(E)检测试剂的灵敏度为6.2 pg/mL,去甲肾上腺素(NE)检测试剂的灵敏度为11.9 pg/mL,多巴胺(DA)检测试剂的灵敏度为4.2 pg/mL,且与多种结构类似物均无明显的交叉反应;E、NE、DA检测试剂的变异系数分别为1.12%~1.77%、1.45%~3.65%、1.30%~1.96%;3种检测试剂的稳定性较好。结论本研究成功建立了检测尿液和血浆中游离E、NE、DA的化学发光免疫检测方法。

Objective To establish a chemiluminescence immunoassay method for free catecholamines in plasma and urine.Methods The plasma or urine samples were extracted,acylated,released and methylated.The processed analytes were treated with solid-phased norepinephrine(NE),adrenaline(E),dopamine(DA)specific antibody reaction,and then added avidin-labeled horseradish peroxidase(HRP),combined with the biotin on the antigen to be tested,could form a solid-phased antibody-antigen-HRP immune reaction complex,HRP catalyzed the substrate to emit light.Results The sensitivity of the established E,NE,DA detection reagent was 6.2,11.9,4.2 pg/mL,respectively,and there was no obvious crossover with many structural analogs;the coefficient of variation of E,NE,and DA detection reagents were 1.12%—1.77%,1.45%—3.65%,1.30%—1.96%,respectively;the stability of the three detection reagents was good.Conclusion This study successfully established a chemiluminescence immunoassay method for detecting free E,NE,and DA in urine and plasma.