高灵敏度人促甲状腺激素(hTSH) 酶联免疫分析试剂盒

以辣根过氧化物酶标记链亲合素 (SA) ,二株识别人促甲状腺激素 (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 ,建立了高灵敏度人促甲状腺激素生物素 亲合素酶联免疫分析 (sTSH BA ELISA)试剂盒的制备方法。本试剂盒的灵敏度为 0 0 2mIU/L ,批内变异系数 <7 8% ,批间变异系数 <9 6% ,回收率为 93 3 %～ 1 0 7 8% ,特异性鉴定显示与其它糖蛋白激素 (如FSH ,HCG ,LH)无明显的交叉反应性。正常参考值范围为 0 3～ 4 1mIU/L。本法所制备的试剂盒与中国原子能科学研究院同位素研究所的TSH IRMA试剂盒的相关方程为 y =1 .2xIRMA 0 1 4,相关系数r =0 969。本试剂盒较常规ELISA灵敏度高、简便、快速 ,适用于临床检测和科研应用

Establishment of an Enzyme Linked Immunosorbent Assay for Human Thyrotropin

A sensitive and specific ELISA for human thyrotropin(hTSH) is established by using two anti hTSH monoclonal antibody. One of them is coated on the microtiter plate, the other is conjugate of biotin. 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 0.02 mIU/L. The intra assay CVs and the intre assay CVs of 3 samples are all lower than 10%. The analytical recovery is in the range from 93.3% to 107.8%. The assay is specific for hTSH and no cross reaction with other glyco proteins, such as hLH, hHCG, hFSH. TSH concentrations range from 0.3 to 4.1 mIU/L in 142 normal subjects. For a reference IRMA(Multipact) method, the correlation regression equation of present method is y=1.2x IRMA 0.14(mIU/L)( r =0.969). This method is a rapid, sensitive, convenient and inexpensive method for measuring hTSH in human serum. It is suitable for clinical and research application.