人血清生长激素化学发光免疫分析方法的建立

摘要

建立链霉亲和素-生物素双抗体夹心化学发光免疫分析法(CLIA)测量人血清生长激素(GH),并进行临床应用检测.以链霉亲和素包被微孔板,生物素化一株GH单克隆抗体(McAb),辣根过氧化物酶标记另一株McAb,校准品与国家标准品进行溯源,建立GH CLIA法,进行性能参数测试,与国外试剂盒比对.结果表明本法线性范围为2.0～150.0μIU/mL,回收率为95.5%,灵敏度0.02μIU/mL,批内、批间CV分别为3.2%～7.2%和3.8%～8.2%,与hPRL、hLH和hTSH叉反应率分别为3.6%、0.05%和0.02%,与hFSH、hCG无交叉反应.本方法与国外同类试剂盒检测187份血清,相关系数为0.9837.37℃,7天热稳定性良好.本方法各项指标均满足临床检测要求,达到国外同类水平,便于临床检验应用.%To develop a streptavidin-biotin chemiluminescent immunoassay based on sandwich way with two anti-growth hormone ( GH) monoclonal antibody ( McAb) for detection of human serum growth hormone. The strep-tavidin coated in microplate, biotinylated McAb and labeled another McAb with horseradish peroxidase, the calibrator traced back to the national standard and parameters of the assay was examined. The developed assay was compared with foreign CH assay kits. The results showed that the linear measurement range of assay was 2.0-150. OμIU/mL, and the recovery rate was 95. 5%. The sensitivity of assay was 0. 02μIU/mL. The intra- andinter-assay CV were 3.2% ~7.2% and 3. 8% ~8. 2% , respectively. The cross reactivity rates with hPRL, hLH and hTSH were 3. 6% , 0. 05% and 0. 02% , respectively. There was no cross reaction with hFSH and hCG. The correlation coefficient between the assay and commercially available CH kit for 187 samples was 0.9837. The developed GH CLIA was stable in 37℃ for 7 days. This assay with simple and fast operation may be suitable for clinical application.