Detection of C-reactive protein utilizing magnetic permeability detection based immunoassays
Detection of C-reactive protein utilizing magnetic permeability detection based immunoassays
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DOI：
10.1021/ac0508649
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发表时间：
2005-09-15
影响因子：
7.4
通讯作者：
Kriz, D
中科院分区：
文献类型：
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作者：
Kriz, K;Ibraimi, F;Kriz, D
描述了一种新的传感技术平台，其在一步分析中集成了磁导率检测和双位点非均相免疫测定。作为一个平台模型，在快速（11.5分钟）高灵敏度（hs）程序中测量C反应蛋白（CRP）（心脏和炎症标志物），检测限低（0.2 mg/L），准确度低（CV = 11%）。在含有固相（多克隆抗CRP偶联二氧化硅微粒）、标记剂（单克隆抗CRP偶联超顺磁性纳米颗粒）和反应缓冲液的1.2 mL一次性试剂小瓶中进行双位点非均相免疫测定。通过使用玻璃毛细管将样品引入试剂小瓶中并手动混合其内容物30 s来测定全血（20 μ L）。在11分钟的沉淀步骤后，将小瓶放入磁导率检测器的线圈中，其测量固相沉淀物中超顺磁性纳米颗粒的富集。磁导率检测和定量基于以下原理：当顺磁性材料置于线圈内时，线圈的电感受到影响。全血患者样本的CRP筛查显示与中心医院hsCRP（y = 1.018x - 0.021，R-2 = 0.980，n = 103）和正常范围CRP（y = 1.02x + 2.53，R-2 = 0.991，n = 33）分析的测量值具有良好的相关性。根据Bland和Altman图，两种方法的平均差异对于hsCRP分析为-0.03 +/- 1.12 mg/L，对于正常范围CRP分析为-3.4 +/- 8.64 mg/L。
A new sensing technology platform integrating magnetic permeability detection and a two-site heterogeneous immunoassay in a one-step analysis is described. As a platform model, measurements of C-reactive protein (CRP), a cardiac and inflammation marker, were performed in a rapid (11.5 min) high-sensitivity (hs) procedure with a low detection limit (0.2 mg/L) and accuracy (CV = 11%). The two-site heterogeneous immunoassay was performed in 1.2-mL disposable reagents vials containing solid phase (polyclonal anti-CRP conjugated silica microparticles), labeling agent (monoclonal anti-CRP conjugated superparamagnetic nanoparticles), and reaction buffer. Whole blood (20 mu L) was assayed by introducing the sample into a reagent vial using a glass capillary and mixing its contents by hand for 30 s. After a 11-min sedimentation step, the vial was placed into the coil of the magnetic permeability detector, which measured the enrichment of superparamagnetic nanoparticles in the solid-phase sediment Magnetic permeability detection and quantification is based on the principle that when paramagnetic materials are placed inside a coil, the inductance of the coil is influenced. Screening of CRP on whole blood patient samples showed good correlation with central hospital measurements for hsCRP (y = 1.018x - 0.021, R-2 = 0.980, n = 103) and normal range CRP (y = 1.02x + 2.53, R-2 = 0.991, n = 33) analyses. The mean differences of the two methods according to the Bland and Altman plots were -0.03 +/- 1.12 mg/L for hsCRP analysis and -3.4 +/- 8.64 mg/L for normal range CRP analysis.