应用时间分辨荧光免疫层析技术建立HBP定量检测的方法

Using Time Resolved Fluorescence Immunochromatography to Establish a Quantitative Detection Method for HBP

摘要: 本文建立了一种肝素结合蛋白(Heparin-binding protein, HBP)的定量检测方法，并检验该方法的性能。主要通过以时间分辨荧光纳米微球作为荧光标记物，结合双抗体夹心法原理和免疫层析技术，实现HBP的定量检测。本检测方法在5~1000 ng/mL线性范围内线性关系良好，方程为y = 0.0324x − 0.0971，R2 = 0.9975。本试剂的批内精密度CV ≤ 5.21%，批间精密度CV ≤ 7.06%。本试剂与中翰盛泰生物技术股份有限公司的肝素结合蛋白测定试剂盒(免疫荧光干式定量法)相关性较好，二者的阳性符合率与阴性符合率分别为96.23%和95.0%，本试剂的假阳性率和假阴性率分别为5.0%和3.77%，诊断符合率为94.52%。本研究建立了HBP定量检测方法，并且该方法具有操作简单、检测快速、结果准确的优点。

Abstract: In this paper, a quantitative detection method for Heparin Binding Protein (HBP) was established and its performance was tested. The quantitative detection of HBP was achieved by using time- re-solved fluorescent nanospheres as fluorescent markers, combined with the principle of double an-tibody sandwich method and immunochromatography technology. The linearity of the detection method was good in the linear range of 5~1000 ng/mL, and the equation was y = 0.0324x − 0.0971, R2 = 0.9975, intra-batch precision CV ≤ 5.21%, inter-batch precision CV ≤ 7.06%. The reagent has a good correlation with Heparin Binding Protein (HBP) Detection Kit (fluorescence immunochroma-tography) developed by Zhonghanshengtai Biotechnology Limited Co., Ltd. The positive coincidence rate and negative coincidence rate were 96.23% and 95.0%, respectively. The false positive rate and false negative rate of this method were 5.0% and 3.77%, respectively, and the diagnostic coin-cidence rate was 94.52%. This study established a quantitative HBP detection method, and the method has the advantages of simple operation, rapid detection and accurate results.