摘要
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.
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 recoverT 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.
出处
《检验医学》
CAS
北大核心
2009年第7期514-517,共4页
Laboratory Medicine
基金
广州市科技攻关项目(2005Z3-E0591)