甲氧基肾上腺素和甲氧基去甲肾上腺素化学发光免疫检测方法的建立及分析性能评估

Establishment and analysis performance evaluation of chemiluminescence immunoassay method for metanephrine and methoxy normetanephrine

目的构建甲氧基肾上腺素(MN)和甲氧基去甲肾上腺素(NMN)的化学发光免疫检测试剂。方法将MN、NMN的特异性抗体分别包被到磁微粒表面,通过标本前处理工艺将待测物生物素化,把与生物素特异性结合的亲和素用辣根过氧化物酶(HRP)标记,通过免疫反应,可以形成固相化的抗体-待测物-生物素-亲和素-HRP免疫反应产物。结果分别建立了MN、NMN的校准曲线,通过性能评估试验测得该检测试剂与多种结构类似物均无明显的交叉反应;MN检测试剂的分析灵敏度为10.21 ng/mL,NMN检测试剂的分析灵敏度为6.91 ng/mL;MN、NMN检测试剂变异系数分别为4.79%~5.25%、3.85%~4.73%;且两种检测试剂的稳定性评估结果显示试剂稳定性较好。结论该研究成功构建了MN、NMN化学发光免疫检测试剂。

Objective To construct a chemiluminescence immunoassay reagent for metanephrine(MN)and methoxy normetanephrine(NMN).Methods The specific antibodies of MN and NMN were respectively coated on the surface of magnetic particles,and the analyte was biotinylated through the sample pretreatment process,and the avidin was labeled with horseradish peroxidase(HRP),through the immune reaction,the immobilized antibody-analyte-biotin-avidin-HRP immune reaction product might be formed.Results The calibration curves of MN and NMN were established.The performance evaluation test showed that there was no obvious cross-reaction between the detection reagent and various structural analogs;the analytical sensitivity of the MN detection reagent was 10.21 ng/mL,and the analytical sensitivity of the NMN detection reagent was 6.91 ng/mL;the coefficients of variation of the detection reagents for MN and NMN were 4.79%-5.25%and 3.85%-4.73%respectively.And the stability evaluation results of the two detection reagents show that the reagents are stable.Conclusion This study successfully constructs the chemiluminescence immunoassay reagent for MN and NMN.