人促甲状腺激素酶促化学发光免疫分析方法的建立

Establishment of Chemiluminescence Immunassay for Thyroid Stimulating Hormone

采用双抗体夹心法建立了定量检测人血清中促甲状腺激素含量的酶促化学发光免疫分析方法，其中，1株抗TSH抗体包被96孔微孔板，另1株与辣根过氧化物酶结合形成酶标记物，鲁米诺-过氧化氢作为发光底物。该方法测量范围为0.1～100 mIU/L ，分析灵敏度为0.04 mIU/L ，批内、批间变异系数分别为3.98％～6.48％和4.61％～13.1％，回收率为95.8％～117.4％，高浓度 TSH样品系列倍比稀释后，测定值与稀释度呈线性相关，相关系数大于0.99。与LH的交叉率＜1.62％，与FSH和 HCG的交叉率分别＜0.05％和＜0.02％。与罗氏化学发光免疫分析方法测定的临床值进行比较，相关性方程为 y＝1.10 x -0.207，相关系数 r＝0.969。与免疫放射分析试剂盒进行比较，相关性方程为 y＝1.00 x＋0.191，相关系数 r＝0.965。本方法操作简便、快速，适用于临床检测和科研应用。

The chemiluminescence immunoassay for thyroid stimulating hormone in human serum was developed . Two monoclonal antibodies of TSH were used in the assay ,one of which was labeled with horseradish peroxidase （HRP） and the other was coated on the microtiter plate .Luminol was used as the substrate of HRP and hydrogen peroxide was introduced into this system . The standard range of the method is 0.1-100 mIU/L . The assay sensitivity is 0.04 mIU/L . The intra- and inter-assay coeffi-cients of variance are 3.98%-6.48% and 4.61%-13.1% .Analytical recovery is 95.8%-117.4% .The correlation coefficient between measured and expected values is more than 0.99 .The cross reaction rate with LH is less than 1.62% ,and that with FSH and HCG is less than 0.05% and 0.02% ,respectively .Compared with determine value clinically in chemiluminescence immunoassay （CLIA ） kit from Roche company ,the correlative equation is y = 1.10 x -0.207 , and correlation coefficient is 0.969 . Compared with immunoradiometric assay （IRMA ） kit ,the correlative equation is y= 1.00 x＋ 0.191 , and correlation coefficient is 0.965 . This method is rapid and convenient , and it is suitable for clinical and research application .