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采用即时检测型生物传感仪测定人血清样本中的降钙素原和白细胞介素-6。

Simultaneous determination of procalcitonin and interleukin-6 in human serum samples with a point-of-care biosensing device.

机构信息

Immunoassays/Immunosensors Lab, Institute of Nuclear & Radiological Sciences & Technology, Energy & Safety, National Centre for Scientific Research "Demokritos", 15341, Aghia, Paraskevi, Greece.

Medicon Hellas S.A., 15344, Gerakas, Greece.

出版信息

Talanta. 2023 Jun 1;258:124403. doi: 10.1016/j.talanta.2023.124403. Epub 2023 Mar 3.

DOI:10.1016/j.talanta.2023.124403
PMID:36889192
Abstract

The simultaneous determination of two inflammatory diseases biomarkers, namely procalcitonin (PCT) and interleukin-6 (IL-6), in human serum samples employing a Point-of-Care device based on Multi Area Reflectance Spectroscopy is presented. Dual-analyte detection was achieved using silicon chips with two silicon dioxide areas of different thickness, one functionalized with an antibody specific for PCT and the other with an antibody specific for IL-6. The assay included reaction of immobilized capture antibodies with mixtures of PCT and IL-6 calibrators with the biotinylated detection antibodies, streptavidin and biotinylated-BSA. The reader provided for the automated execution of the assay procedure, as well as for the collection and processing of the reflected light spectrum, the shift of which is correlated to analytes concentration in the sample. The assay was completed in 35 min and the detection limits for PCT and IL-6 were 2.0 and 0.01 ng/mL respectively. The dual-analyte assay was characterized by high reproducibility (the intra- and inter-assay coefficients of variation were less than 10% for both analytes) and accuracy (the percent recovery values ranged from 80 to 113% for both analytes). Moreover, the values determined for the two analytes in human serum samples with the assay developed were in good agreement with the values determined for the same samples by clinical laboratory methods. These results support the potential of the proposed biosensing device application for inflammatory biomarkers determination at the Point-of-Need.

摘要

基于多区域反射光谱的即时检测设备同时测定两种炎症性疾病生物标志物,即降钙素原(PCT)和白细胞介素-6(IL-6),在人血清样本中的应用。双分析物检测是通过具有两个不同厚度二氧化硅区域的硅芯片实现的,一个区域功能化有针对 PCT 的抗体,另一个区域功能化有针对 IL-6 的抗体。该测定法包括固定化捕获抗体与 PCT 和 IL-6 校准品混合物的反应,然后与生物素化检测抗体、链霉亲和素和生物素化-BSA 结合。阅读器提供了用于自动执行测定程序以及收集和处理反射光光谱的功能,其光的偏移与样品中分析物的浓度相关。该测定在 35 分钟内完成,PCT 和 IL-6 的检测限分别为 2.0 和 0.01ng/mL。双分析物测定法具有高重复性(两种分析物的批内和批间变异系数均小于 10%)和准确性(两种分析物的回收率值范围为 80%至 113%)。此外,用所开发的测定法测定人血清样本中的两种分析物的结果与用临床实验室方法测定相同样本的结果非常吻合。这些结果支持了所提出的生物传感设备在即时检测点用于炎症性生物标志物测定的应用潜力。

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