Establishment of Magnetic Microparticles-Assisted Time-Resolved Fluoroimmunoassay for Determinating Biomarker Models in Human Serum

In order to early screen and detect suspected biomarkers from pathogens and the human body itself, tracers or reaction strategies that can act as signal enhancers have been proposed forth at purpose. In this paper, we discussed the applicability of magnetic microparticles-assisted time-resolved fluoroimmunoassay (MMPs-TRFIA) for sensitive determination of potential analytes. Hepatitis B e antigen, antibody to hepatitis B surface antigen and free triiodothyronine were used as biomarker models to explore the reliability of the method. By coupling with bioprobes, MMPs were used as immunoassay carriers to capture target molecules. Under optimal condition, assay performance, including accuracy, precision and specificity, was outstanding and demonstrated satisfactory. To further evaluate the performance of the MMPs-TRFIA in patients, a total of 728 serum samples from hospital were analyzed for three biomarkers in parallel with the proposed method and chemiluminescence immunoassay kit commercially available. Fairly good agreements are obtained between the two methods via data analysis. Not only that but the reliability of MMPs-TRFIA has also been illustrated by three different reaction models. It is confirmed that the novel method modified with MMPs has been established and showed great potential applications in both biological detection and clinical diagnosis, including big molecule protein and low molecular weight haptens.为实现从病原体及人体自身组分中早期筛查、检测疑似生物标志物的目标，学界已提出可作为信号增强剂的示踪剂或反应策略。本文探讨了磁微粒辅助时间分辨荧光免疫测定（magnetic microparticles-assisted time-resolved fluoroimmunoassay，MMPs-TRFIA）用于潜在分析物灵敏检测的适用性。本研究以乙型肝炎e抗原、乙型肝炎表面抗原抗体及游离三碘甲状腺原氨酸作为生物标志物模型，对该方法的可靠性开展验证。通过与生物探针偶联，磁微粒可作为免疫测定载体捕获靶标分子。在最优实验条件下，该方法的检测性能（包括准确度、精密度与特异性）表现优异，结果令人满意。为进一步评估MMPs-TRFIA在临床患者样本中的检测性能，本研究共纳入728份医院采集的血清样本，采用所提方法与市售化学发光免疫分析试剂盒并行检测三种目标生物标志物。数据分析结果显示，两种检测方法间具有良好的一致性。不仅如此，通过三种不同的反应模型，进一步验证了MMPs-TRFIA的可靠性。研究证实，本研究所构建的磁微粒修饰新型检测方法，在生物检测与临床诊断领域（涵盖大分子蛋白与小分子半抗原）均展现出可观的应用潜力。