摘要
目的利用时问分辨荧光免疫分析（TrFIA）技术，建立新生儿促甲状腺素（NeonatalTSH）微量检测法，并研制其检测试剂盒。方法采用一株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＋），固相双抗体夹心时间分辨荧光免疫分析法检测NeonatalTSH。结果自建的NeonatalhTSHTrFIA法灵敏度高，可达2．28btU／ml，且在测量范围内，自制试剂盒剂量一反应曲线的相关系数r达0．9987，与hLH，hFSH，HCG无明显交叉反应。分析内或分析间质控结果在靶值±25％内，室内低质控、高质控分析内和分析间变异系数（Cy）分别为7．12％和6．09％，7．82％和7．03％。与DELFIAONeonatalTSH试剂标准品进行准确性比对，实测值与标示效价比平均值为1．05。与DELFIA睁NeonatalTSH试剂比对试验符合率一致，相关系数r可达0．9822。试剂盒室间质评能力比对成绩100％。结论自建的NeonatalTSHTrFIA法具有灵敏度高，特异度强，准确度高，精密性好和非放射性等优点，具有良好的临床应用价值。
Objective To establish a method of Time-resolved Fluoroimmunoassay（TrFIA） for the quantitative determina- tion 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 its 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 μU/ml. The correlation coef- ficient of self-made kit＇s curve was 0. 998 7. There was no obvious cross-reaction to human luteinizing hormone（hLH） ,hu- man 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 cor- relation coefficient of self made Kit and DELFIA Neonatal TSH kit was 0. 982 2,and the results of self made Kit and DELFIA~ Neonatal TSH kit were showing no difference. The lab＇s proficiency testing was excellent. Conclusions The self-made Neonatal TSH TrFIA has several advantages including high sensitivity, strong specificity,good accuracy, nice precision,and non-radioactive immunoassay. It is suitable for clinical use.
出处
《现代检验医学杂志》
CAS
2009年第4期79-81,共3页
Journal of Modern Laboratory Medicine
基金
基金项目：科技型中小企业技术创新基金（08c26114411281）,广州市科技计划项目〈2006V42C0051）.
