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基于电化学微流控微马达的免疫分析法用于检测疑似脓毒症早产儿样本中的 C 反应蛋白

Electrochemical Microfluidic Micromotors-Based Immunoassay for C-Reactive Protein Determination in Preterm Neonatal Samples with Sepsis Suspicion.

机构信息

Department of Analytical Chemistry, Physical Chemistry and Chemical Engineering, Universidad de Alcalá, Ctra. Madrid-Barcelona, Km. 33.600, 28871 Alcalá de Henares, Madrid, Spain.

Chemical Research Institute "Andrés M. Del Rio", Universidad de Alcalá, 28871 Alcalá de Henares, Madrid, Spain.

出版信息

Anal Chem. 2020 Apr 7;92(7):5048-5054. doi: 10.1021/acs.analchem.9b05384. Epub 2020 Mar 10.

Abstract

Online coupling of a micromotor-based immunoassay and a microfluidic electrochemical detection was explored as a new approach for C-reactive protein (CRP) determination in serum and preterm neonatal plasma samples with sepsis suspicion. The approach combines the advantages of micromotors (self-fluid mixing capabilities leading to a faster assay in low sample volumes) and electrochemical microfluidic (flow-controlled ultraminiaturized electrochemical detection, high sensitivity, and low-cost) technologies. Both technologies elegantly meet the point of care testing or bed side device requirements such as low analysis times, miniaturization and simplification, and single use. Anti-CRP functionalized micromotors (anti-CRP-rGO(reduced graphene oxide)/Ni/PtNPs (platinum nanoparticles))-based immunoassay coupled to thin layer Au-based electrochemical microfluidics operating at -0.20 V under controlled fluidic detection operations (30 μL min) allowed the sensitive (LOD = 0.54 μg/mL) and accurate CRP determination using very low volume preterm neonatal clinical samples (<10 μL) in just 8 min of total assay time. These excellent analytical characteristics obtained linked to the full automatization of the immunoassay allowed the fast and accurate determination of CRP in hardly available clinical samples as those coming from preterm infants with suspected sepsis. These results demonstrated the usefulness of the approach which meets the clinical requirements as a future point-of-care device for clinical analysis.

摘要

在线耦合基于微电机的免疫测定和微流控电化学检测被探索作为一种新的方法来确定 C 反应蛋白 (CRP) 在血清和疑似败血症的早产儿血浆样本中。该方法结合了微电机的优点 (自液混合能力,导致在低样品量的更快的测定) 和电化学微流控 (流量控制超小型化电化学检测、高灵敏度和低成本) 技术。这两种技术都巧妙地满足了即时检验或床边设备的要求,如分析时间短、微型化和简化以及一次性使用。基于抗 CRP 功能化微电机 (抗 CRP-rGO(reduced graphene oxide)/Ni/PtNPs (铂纳米粒子))的免疫测定与基于薄层金的电化学微流控相结合,在受控的流体检测操作下(-0.20 V)运行,允许使用非常低体积的早产儿临床样本(<10 μL)在 8 分钟的总测定时间内进行敏感(LOD = 0.54 μg/mL)和准确的 CRP 测定。这些优异的分析特性与免疫测定的完全自动化相结合,使得能够快速准确地测定来自疑似败血症的早产儿的几乎无法获得的临床样本中的 CRP。这些结果证明了该方法的有用性,作为未来的即时检测设备,满足临床分析的要求。

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