促卵泡激素酶促化学发光免疫分析试剂盒的研制

Development of Chemiluminscence Immunoassay Kit for Follicle-stimulating Hormone

采用双抗体夹心法建立了测定人血清FSH酶促化学发光免疫分析方法。本方法的测量范围为1.5～100 IU/L,灵敏度为0.36 IU/L,批内变异系数为1.0%～1.6%,批间变异系数为4.0%～4.6%,回收率为90.6%～117.8%,稀释实验测定值与稀释度呈线性相关,相关系数r=0.990。与TSH的交叉率<0.23%,与LH和HCG的交叉率分别<0.16%和<0.27%。与贝克曼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=0.85x-0.03,相关系数r=0.954。与免疫放射分析试剂盒进行比较,相关性方程为y=0.97x+5.29,相关系数r=0.965,与放射免疫分析试剂盒,进行比较,相关性方程为y=0.95x-3.82,相关系数r=0.954。方法学鉴定结果符合免疫分析的基本要求。

The chemiluminescence immunoassay for follicle-stimulating hormone in human serum was developed. Two monoclonal antibodies of FSH were used in the assay, one of which was labeled with horseradish peroxidase（HRP） and the other was coated on the microtite plate. Luminol was used as the substrate of HRP and hydrogen peroxide was introduced into this system. The standard range of the method was 1.5-100 IU/L. The assay sensitivity was 0.36 IU/L. The intra- and inter-assay coefficients of variance were 1.0%- 1.6% and 4.0%-4.6% respectively. Analytical recovery was 90.6%-117.8%. The correletion coefficient between measured and expected values was 0. 990. The crossreacting rate with TSH was less than 0.23%, and that with LH and HCG less than 0.16% and 0.27%. Compared with determine value clinically in chemiluminescence immunoassay （CLIA） kit from Beckman eoulter company, the correlative equation was y=0.85x-0.03, and correlation coefficient was 0. 954. Compared with immunoradiometric assay （IRMA） kit and radioimmunoassay（RIA） kit, the correlative equation were y = 0. 97x-1- 5. 29 and y = 0. 95x-3.82, and their correlation coefficient were 0. 965 and 0. 954 respectively.