滤纸干血-新生儿促甲状腺素酶联免疫分析试剂盒的研制

The Establishment of an Enzyme Linked Immunosorbent Assay for Neonatal Thyrotropin

摘要: 以辣根过氧化物酶标记链亲合素(SA),两株识别人促甲状腺素(hTSH)高度特异的单克隆抗体分别用于包被微孔和生物素化,以四甲基联苯胺(TMB)为底物,建立了滤纸干血-新生儿促甲状腺素生物素-亲合素酶联免疫分析(NeonatalTSH-BA-ELISA)试剂盒的制备方法。本方法的灵敏度为0.5mIU/L,批内变异系数<15%,批间变异系数<20%,回收率为90.1%～103.3%。本方法所制备的试剂盒与NeonatalTSH-IR-MA(磁颗粒法)试剂盒具有良好的相关性,相关方程为y=1.01x+0.34,相关系数为r=0.873。本试剂盒无放射性,操作简便、快速,适用于临床检测和科研应用。

Abstract: A sensitive and specific ELISA for neonatal thyrotropin(Neonatal TSH) is established by using two antiTSH monoclonal antibodies. One of them is coated on the microtiter plate, and the other is biotinlated. The label is horseradish peroxidase(HRP) conjugate of streptavidin. TMBH2O2 solution is used as the substrate of HRP. The sensitivity of the assay is 0.5 mIU/L. The intraand introassay CVs of 3 samples are lower than 15% and 20% respectively. The analytical recovery is in the range from 90.1% to 103.3%. Compared with IRMA, the correlative equation is y=1.01x+0.34, and r=0.873. This method for measuring neonatal TSH is rapid, sensitive, convenient and inexpensive. It is suitable for clinical and research application.