摘要
目的建立血清胰岛素(Ins)时间分辨荧光免疫分析法(time-resolved fluoroimmunoassay,TRFIA)。方法采用一株Ins-MAb包被于固相微孔板上,另一株Ins-MAb用DTTA-Eu3+标记作为示踪剂采用Victor 1420荧光仪测定,。并对本法进行评价。结果本法的标准曲线范围为5～160 mU/L。灵敏度为0.35mU/L。检测3个浓度的Ins样品,批内CV为0.90%～4.10%,批间CV为4.55%～4.96%。平均回收率为97.7%～102.9%。用本法与DSL酶免分析方法同时测定80份血清样品,相关系数为0.982。结论本文建立的Ins-TRFIA具有特异性强、灵敏度高及操作简单等优点,适用于临床血清Ins的检测。
Objective A time-resolved fluoroimmunoassay （TRFIA） for the measurement of insulin （Ins） in human serum had been established. Methods The test was based on the one-step sandwich technique with two MAbs directed against two different epitopes of Ins using coated 96-well mierotitre plate as the solid phase and DITA-Eu3＋ as the label, and the established method was evaluated with Victor 1420 muhilabel counter. Results The standard ranges were 5 - 160mU/L. The detection limit was 0.35mU/L. At three different levels of Ins, the intra-assay CV was 0.90% to 4.10%, the inter-assay CV was 4.55% to 4.96%. The average re- covery rates were 97.7% - 102.9%. The detected results with this method closely corresponded to those of by ELISA with DSL （R = 0. 982, n = 80）. Conclusion The established TRFIA in this study was specific, sensi- tive, easy to use, and suitable for clinical serum insulin measurement.
出处
《标记免疫分析与临床》
CAS
2012年第1期36-39,共4页
Labeled Immunoassays and Clinical Medicine
