Gutiérrez-Capitán Manuel, Balada Eva, Aviñó Anna, Vilaplana Lluïsa, Galve Roger, Lacoma Alícia, Baldi Antonio, Alcamí Antonio, Noé Véronique, Ciudad Carlos J, Eritja Ramón, Marco María-Pilar, Fernández-Sánchez César
Instituto de Microelectrónica de Barcelona (IMB-CNM) CSIC, 08193 Bellaterra, Spain.
Institute for Advanced Chemistry of Catalonia (IQAC) CSIC, 08034 Barcelona, Spain.
Anal Chem. 2025 Jun 10;97(22):11863-11873. doi: 10.1021/acs.analchem.5c01605. Epub 2025 May 30.
Providing viral load numbers of infection events aids in the identification of disease severity and in the effective overall patient management. Gold-standard polymerase chain reaction (PCR) techniques make this possible but cannot be applied at the point of need and in low-resource settings. Here, we report on the development of a compact analytical platform that can detect a conserved sequence of the RNA of severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) in 40 min in nasopharyngeal swab samples without the need for any previous purification or gene amplification steps. It combines electrochemical and paper fluidic approaches together with a sandwich hybridization assay performed on magnetic nanoparticles (MNPs) modified with a tailor-designed capture DNA hairpin. The device proves to quantitatively detect viral RNA in a retrospective study carried out with nasopharyngeal swab samples. A sensitivity of 100% and a specificity of 93% were estimated by the receiver operating characteristic (ROC) analysis. However, although molar concentration values of the target RNA sequence are provided, these estimates do not fully correlate with the viral load numbers estimated by RT-qPCR over the whole Ct sample range. Empirical studies have been carried out that have provided clear insights into this hurdle and simple solutions to overcome it, without depriving the device of the features required for potential use in a point-of-care (PoC) environment.
提供感染事件的病毒载量数值有助于确定疾病严重程度并有效管理患者整体情况。金标准聚合酶链反应(PCR)技术使这成为可能,但无法在需要时和资源匮乏的环境中应用。在此,我们报告了一种紧凑型分析平台的开发,该平台可在40分钟内检测鼻咽拭子样本中严重急性呼吸综合征冠状病毒2(SARS-CoV-2)RNA的保守序列,无需任何先前的纯化或基因扩增步骤。它将电化学和纸流体方法与在经定制设计的捕获DNA发夹修饰的磁性纳米颗粒(MNP)上进行的夹心杂交分析相结合。在一项对鼻咽拭子样本进行的回顾性研究中,该设备被证明能够定量检测病毒RNA。通过受试者工作特征(ROC)分析估计灵敏度为100%,特异性为93%。然而,尽管提供了目标RNA序列的摩尔浓度值,但这些估计值与整个Ct样本范围内通过RT-qPCR估计的病毒载量数值并不完全相关。已经开展了实证研究,对这一障碍有了清晰的认识,并提出了克服它的简单解决方案,同时又不剥夺该设备在即时检测(PoC)环境中潜在使用所需的特性。
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