摘要
目的研制一种人游离甲状腺素(FT4)时间分辨荧光免疫分析法(TRFIA)试剂盒。方法采用直接竞争法建立FT4检测方法(固相包被T4-复合物,与样本中的FT4竞争结合Eu标记的T4单克隆抗体),并对该试剂盒各项性能指标进行评价。结果该试剂盒的分析灵敏度为不高于2.0pmol/L,工作范围为2.0～120.0pmol/L。分析内变异系数为4.4%～5.7%、分析间变异系数为6.5%～8.2%,其总变异系数小于9.0%。与T3、FT3、rT3和rT4的交叉反应率分别为:1.12%、0.34%、0.56%和1.51%。稳定性试验表明该试剂盒可以在2～8℃稳定1年,37℃稳定7d。用该试剂盒对500份健康人血清标本进行测试,统计该试剂盒的正常参考值范围是11.5～22.7pmol/L。200份血清标本用本试剂盒与国外的化学发光试剂盒同时检测FT4,其相关系数为0.993。结论该试剂盒各项指标均达到临床检测要求,可替代国外化学发光法的同类产品试剂盒。
Objective To prepare a kit for the detection of human Free Thyroxine （FT4） in serum by time resolved fluoroimmunoassay （TRFIA）. Methods Competitive reaction method was used to build an method of TRFIA for the detection of FT4 （solid phase was coated with T4 complex,and competitively binded to Eu labeled T4 monoclonal antibody with FT4）. And the performance of the kit was evaluated. Results The analytical sensitivity was not higher than 1.5 pmol/L,and the working range of the kit was 2.0--120.0 pmol/L. The analytical sensitivity was 1.5 pmol/L. The intra and inter assay coefficients of variation were 4.4 %- 5.7% and 6.5 %- 8.2%, respectively. The percentage of cross reactivity with T3,FT3, rT3 and rT4 was 1.12 % ,0.34 % ,0.56 % and 1.51 %, respectively. The TRFIA FT4 reagents can be stored at 2--8 %C for one year and at 37 %C for 7 days. The cutoff value for TRFIA FT4 kit in healthy subjects （n= 500） was 11.5--22.7 pmol/L,in according to 500 cases of health sample serum with this kit. The correlation coefficient of blood samples detection results （n= 185） between home made and foreign commercially available CLIA FT4 kit （SIEMENS） was 0. 993. Conclusion FT4 TRFIA kit was a valuable diagnostic kit for clinical application with better sensitivity, specificity,and accuracy.
出处
《检验医学与临床》
CAS
2012年第10期1203-1205,共3页
Laboratory Medicine and Clinic
