Low-Cost Full-Range Detection of C-Reactive Protein in Clinical Samples by Aptamer Hairpin Probes and Coprecipitation of Silver Ions and Gold Nanoparticles
Low-Cost Full-Range Detection of C-Reactive Protein in Clinical Samples by Aptamer Hairpin Probes and Coprecipitation of Silver Ions and Gold Nanoparticles
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DOI：
10.1021/acs.analchem.3c01131
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发表时间：
2023-08-02
影响因子：
7.4
通讯作者：
Liu，Lihong
中科院分区：
文献类型：
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作者：
Yan，Hong;Hu，Xiumei;Liu，Lihong
C-反应蛋白（CRP）水平可能与不同的疾病背景有很大关系。然而，昂贵的抗体阻碍了基于抗体的全范围CRP测定的临床实用性，特别是在发展中国家。本研究将金纳米粒子（AuNPs）、银（AgI）、曙红Y（Eosin Y）和具有Ag+介导的胞嘧啶-胞嘧啶错配的适体发夹探针（AHP）结合起来，建立了一种低成本、无抗体、96孔板全范围CRP检测方法，即Au@AgI/Eosin Y-AHP方法。在结合目标CRP后，AHP释放Ag+，其随后诱导AuNPs在AgI胶体表面上聚集，导致AuNPs表面对曙红Y的吸附显著增加。在0.01-40 ng/mL的范围内，不含和含CRP的上清液中曙红Y的荧光强度（FI）的变化与CRP的浓度成正比（r= 0.9969），96个样品可以在45分钟内被酶标仪同时检测到。值得注意的是，用Au@AgI/Eosin Y-AHP法测定的100份临床样本的CRP水平与乳胶增强免疫比浊法测定的CRP水平具有良好的相关性（r= 0.986）。此外，基于Au@AgI/曙红Y-AHP方法的试剂盒100次测试仅需8.1美元。因此，这种新方法对于那些昂贵的检测方法负担不起的欠发达地区是有益的。
C-reactive protein (CRP) levels can vary widely related to diverse disease contexts. However, expensive antibodies have impeded the clinical utility of antibody-based full-range CRP assays, especially in developing countries. Herein, we established a low-cost, antibody-free, 96-well plate-based full-range CRP detection method by combining gold nanoparticles (AuNPs), silver iodide (AgI), Eosin Y, and the aptamer hairpin probe (AHP) with Ag+-mediated cytosine–cytosine mismatches, that is, the Au@AgI/Eosin Y-AHP method. After binding the target CRP, the AHP released Ag+, which subsequently induced the aggregation of AuNPs on the surface of AgI colloids, resulting in a significant increase in the adsorption of Eosin Y on the surface of AuNPs. The changes in fluorescence intensity (FI) of Eosin Y in the supernate without and with CRP were proportional to the concentration of the CRP in the wide range of 0.01–40 ng/mL (r= 0.9969), and 96 samples can be detected in 96-well plates simultaneously by a microplate reader within 45 min. Remarkably, the CRP levels of 100 clinical samples achieved with the Au@AgI/Eosin Y-AHP had a good correlation with those obtained with the latex-enhanced immune turbidimetry assay (r= 0.986). Furthermore, the kit based on the Au@AgI/Eosin Y-AHP method costs only $8.1 for 100 tests. Therefore, the new method is beneficial for less developed areas where expensive assays are not affordable.