血清叶酸化学发光免疫检测技术及试剂盒的研发

Reasearch and development of chemiluminescent immunoassay for human serum folic acid

研究建立了一种叶酸化学发光免疫检测技术,并开发成检测试剂盒,能快速、准确检测血清中叶酸的含量。首先用样本处理剂快速将血清中结合态叶酸完全释放,再用高亲和力的叶酸结合蛋白进行检测,建立一种竞争化学发光免疫分析方法。通过对试剂盒中各组分的优化,试剂盒最低检测限为1 ng/mL,线性测量范围为1.5～50 ng/mL,批内及批间变异系数(CV%)均小于10%。稳定性好,37℃加速稳定性能维持10 d, 2℃～8℃能稳定维持为16个月。通过对200份临床血清样本进行检测,试剂盒检测结果与国外著名试剂盒厂家(贝克曼)的相关性系数(R)可达到0.9847。该叶酸检测技术创新地运用了生物素-亲和素反应体系,改变了传统抗原抗体反应存在不均一不稳定等弊端,提高了检测的准确度和灵敏度,并研制出新的样本处理试剂和方法,使血清中叶酸释放更加迅速和完全,提高了试剂盒的准确度并降低了成本。试剂盒的各项性能指标均居于国内领先水平且成本较低,检测量大,可替代国外同类型检测试剂盒。

The study developed a chemiluminescent immunoassay(CLIA)test kit which can detect folic acid in serum quickly and accurately.First of all,the sample treatment agent was used to rapidly release the bounding folate in the serum completely,and then folate was detected with folate binding protein to establish a competition chemiluminescence immunoassay.Through the optimization of each component in the kit,the minimum detection limit of this kit was 1 ng/mL,the linear range was 1.5-50 ng/mL with high specificity,and the intra-assay and inter-assay coefficients of variation were both less than 10%.Its storage duration could be 16 months at 2-8℃.Two hundred clinical serum samples were tested,and the correlation coefficient of the kit test results with the control kit manufacturers(Beckman)was 0.9847.The detection technology innovatied the use of Biotin-Avidin-System to improve the detection accuracy and sensitivity,and developed a new sample processing reagents and methods to make the release of serum folic acid more quickly and completely.All the technical parameters of the folic acid quantification kit are setting the pace in the home market.It can be widely used in the detection of clinical samples.