C-肽时间分辨荧光免疫分析试剂盒的研制

目的研制血清C-肽的时间分辨荧光免疫分析（TRFIA）试剂盒。方法采用双抗体夹心法建立C-肽TRFIA试剂盒，并进行方法学评价。结果C-肽可测量范围为0．4-20ng／mL；分析敏感性为0．23ng／mL；分析批内、批间精密度分别为7．99％、11．97％；C-肽回收率为99．57％；与胰岛素和胰岛素原无交叉反应。稳定性试验表明，试剂可以贮存在4℃稳定9个月，37℃稳定7d；该试剂盒测试200份正常人血清样本，其参考范围为0．358-3．590ng／mL；收集122份血清样本，用该试剂盒与国外其他方法的同类试剂盒同时检测，其相关系数为0．9432，线性方程为Y=0．9854X＋0．5063。结论C-肽TRFIA试剂盒各项指标均达到临床检测要求，适合临床推广使用．

Development of a time-resolved fluoroimmunoassay kit for detecting C-peptide

Objective To develop a time-resolved fluoroimmunoassay(TRFIA) kit for detection of C-peptide in serum. Methods Two anti-C-peptide monoclonal antibodies were used to develop the sandwich TRFIA kit for detection of C-peptide in serum and its methodology was evaluated. Results The detection range of C-peptide was 0.4-20 ng/mL.The analytical sensitivity was 0.23 ng/mL. The intra-assay and inter-assay coefficients of variation were 7.99% and 11.97% respectively. The recovery rate was 99.57%. There was no cross-reactivity with insulin and proinsulin. The TRFIA C-peptide reagents could be stored at 4 ℃ for 9 months and at 37 ℃ for 7 days. The reference value for TRFIA C-peptide kit in healthy subjects(200 cases) was 0.358-3.590 ng/mL. The correlation coefficient of blood samples detection results(122 cases) between this TRFIA kit and commercially available electrochemiluminescense immunoassay C-peptide kits was 0.943 2. The linear equation was Y=0.985 4X+0.506 3. Conclusions The developed TRFIA C-peptide kit is valuable for clinical application with good sensitivity, precision, accuracy, specificity and stability.