新生儿促甲状腺素的时间分辨荧光免疫分析及其试剂盒研制.doc.doc

新生儿促甲状腺素的时间分辨荧光免疫分析及其试剂盒研制 谭玉华，冯健明，卢顺舵，罗建安，李建国，季涛，李贵情 （广州市丰华生物工程有限公司，广州 510730 ） 摘要：目的 利用时间分辨荧光免疫分析（TrFIA）技术，建立新生儿促甲状腺素（Neonatal TSH）微量检测法，并研制其检测试剂盒。方法 采用了一株TSH单克隆抗体用于固相包被，另一株TSH单克隆抗体用于标记铕（Eu3+），固相’s Test Reagent Tan Yu-hua,Feng Jian-ming, Lu Shun-duo, Luo Jian-an, Li Jian-guo, Ji Tao,Li Gui-qing ( Guang zhou Fenghua Bio-engineering Co.,LTD,Guangzhou 510730 China) Abstract: Objective To establish a method of Time-resolved Fluoroimmunoassay(TrFIA) for the quantitative determination of human thyroid stimulating hormone(hTSH) in blood specimens dired on filter paper as an aid in screening newborns for neonatal hypothyroidism,and to prepare it抯 test reagent .Methods The method based on solidphase sandwich TrFIA for Neonatal Hypothyropin(Neo-hTSH),which used two anti-TSH monoclonal antibody -one was coated on microplates and another was labeled by Europium. Results The assay sensitivity of the method was 2.28礥/ml.The correlation coefficient of self made kit抯 curve was 0.9987.There was no obvious cross-reaction to human luteinizing hormone(hLH) , human follicle stimulating(hFSH) and human chorionic gonadotropin(HCG).The intra-assay and the inter-assay measured values of controls were not more than 25% difference from their labeled values, still their intra-assay and the inter-assay coefficients of variation(CV) of low and high controls were 7.12% and 6.09%, 7.82% and 7.03%,respectively. The correlation coefficient of self made Kit and DELFIA ?Neonatal TSH kit was 0.9822,and the results of self made Kit and DELFIA ?Neonatal TSH kit were showing no difference. The lab抯 proficiency testing was excellent. Conclusions The Neonatal TSH TrFIA established which has several advantages including high sensitivity, strong specificity, good accuracy, nice precision, and non-radioactive immunoassay. It is suitable for clinical use. Key words: Neonatal; Hypothyropin; Time-resolved Fluoroimmunoassay 新生儿血液中促甲状腺素（Neonatal TSH）浓度升高(出生72小时后采血)是原发性甲状腺功能低下症（CH）最早的实验指标。CH是造成儿童智力