Development of The Dual-labeling Time-resolved Fluoroimmunoassay for Detection of C-peptide and Insulin and Its Initially Application
Development of The Dual-labeling Time-resolved Fluoroimmunoassay for Detection of C-peptide and Insulin and Its Initially Application
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DOI：
10.3724/sp.j.1206.2011.00037
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发表时间：
2011
影响因子：
0.3
通讯作者：
M. Qiang
中科院分区：
文献类型：
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作者：
M. Qiang
为了同时检测人血清中的c肽和胰岛素，初步建立了双标记时间分辨荧光免疫法（TRFIA）检测c肽和胰岛素的方法。将两种捕获的单克隆抗体混合在一起，然后包被在人血清中。两种对应的示踪单克隆抗体分别用Eu3+（抗胰岛素）和Sm3+（抗c肽）螯合物标记。采用双抗体夹心一步法建立了同时检测人血清中c肽和胰岛素的TRFIA。结果表明，C肽分析的灵敏度可达0.2 μg/L，线性范围为0.5~22 μg/L，回收率为99.6%，检测内变异系数为4.6%~6.0%，检测间变异系数为5.1%~7.6%；胰岛素分析的灵敏度为0.8 mU/L，线性范围为3.6~180 mU/L，回收率为99.4%，试验内变异系数为3.7%~6.0%，试验间变异系数为5.1%~8.0%。同时采用自制和市售c肽和胰岛素试剂盒（PerkinElmer）检测200份样品。相关系数分别为0.98和0.99。综上所述，自制的同时检测c肽和胰岛素的双标记时间分辨荧光免疫法可能是一种简便、灵敏、快速的方法，可用于胰岛素分泌不足引起的糖尿病的快速诊断，也为临床应用中c肽和胰岛素的血清学高筛选提供了另一种途径。
To simultaneously detecting the C-peptide and insulin in human serum,a dual-labeling time-resolved fluoroimmunoassay(TRFIA) for detection of C-peptide and insulin was preliminary founded.Two capture monoclonal antibodies were mixed together and then coated in human serum.Two counterpart tracer monoclonal antibodies,were labeled with Eu3+(anti-Insulin) and Sm3+(anti-C-peptide) chelates,respectively.The double antibody sandwich in one step method was used to develop the TRFIA for simultaneously detection of the C-peptide and insulin in human serum.These results showed that sensitivity of C peptide analysis was up to 0.2 μg/L,linear range is 0.5～22 μg/L,the recovery percent is 99.6%,The intra-and inter-assay coefficient of variation were 4.6%～6.0% and 5.1%～7.6%,respectively;sensitivity of insulin analysis was up to 0.8 mU/L,linear range is 3.6～180 mU/L,the recovery percent is 99.4%,the intra-and inter-assay coefficient of variation were 3.7%～6.0% and 5.1%～8.0%,respectively.Moreover,200 samples were tested by the self-made and commercially available C-peptide and insulin kit(PerkinElmer) kits at the same time.The correlation coefficient was 0.98 and 0.99,respectively.In conclusion,the self-made dual-labeling time-resolved fluoroimmunoassay for simultaneously detection of C-peptide and insulin might be a simple,sensitive and rapid method for rapid diagnosis for the diabetes caused by the lack of insulin secretion,it would also provide an alternative route for serology high-screening of the samples for C-peptide and insulin in clinical application.