游离三碘甲状腺原氨酸(FT3)时间分辨荧光免疫分析试剂盒的研制

Development of the time-resolved fluoroimmunoassay kit for detecting FT3

目的研制一种游离三碘甲状腺原氨酸(free triiodothyronine,FT3)时间分辨荧光免疫分析(time-resolved fuoroimmunoassay,TRFIA)试剂盒。方法采用直接竞争法建立游离三碘甲状腺原氨酸(FT3)检测TRFIA试剂盒(固相包被T3-复合物,与样本中的FT3竞争结合DTTA-Eu3+标记的T3单克隆抗体),并对试剂盒各项指标进行评价。结果该试剂盒的有效检测浓度范围为1.5~70 pmol/L,分析灵敏度为不高于0.7 pmol/L,批内、批间的不精密度分别不高于8.2%与10.5%。与T4、FT4、反T3、反T4交叉反应率分别为:<0.01%、0.11%、<0.01%、<0.01%。稳定性试验表明试剂可以在2~8℃稳定一年,37℃稳定7d。用该试剂盒对500份正常人血清标本进行测试,统计该试剂盒的正常参考值范围是3.2~6.5 pmol/L。156份血清标本用本试剂盒与国外某知名品牌化学发光法试剂盒同时检测,其相关系数为0.992。结论试剂盒各项指标均达到临床检测要求,可考虑作为国外化学发光法同类产品试剂盒的替代产品。

Objective To develop a kit for the detection of human free triiodothyronine（FT3） in serum by time-resolved fuoroimmunoassay（TRFIA）.Methods The TRFIA FT3 kit was established by direct competitive method（solid-phase was coated with T3 complex,and competitively binded to T3 monoclonal antibody labeled DTTA-Eu3＋with FT3）.And the performance of the kit was evaluated.Results The effective working range of the kit was 1.5~70 pmol/L and the analytical sensitivity was not higher than 0.7 pmol/L.The intra-assay and inter-assay coeffcients of variation were not higher than 8.2% and 10.5%,respectively.The percentage of cross reactivity with T4,FT4,rT3,rT4 were 0.01%,0.11%,0.01% and 0.01%,respectively.The TRFIA FT3 reagents can be stored at 2℃ ~8℃ for one year and at 37℃ for 7 days.The cutoff value for TRFIA FT3 kit in healthy subjects was 3.2 pmol/L^6.5 pmol/L,in according to 500 cases of health sample serum with this kit.The correlation coeffcient of blood samples detection results（n=156） between home-made and CLIA kit of one well-known foreign brands was 0.992.Conclusion FT3 TRFIA kit was a valuable diagnostic kit for clinical application with better sensitivity,specifcity,and accuracy.