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使用免疫传感器检测刺突蛋白的电阻抗光谱法诊断 SARS-CoV-2 感染。

Diagnostics of SARS-CoV-2 infection using electrical impedance spectroscopy with an immunosensor to detect the spike protein.

机构信息

São Carlos Institute of Physics (IFSC), University of São Paulo (USP), 13566-590, São Carlos, Brazil.

Nanotechnology National Laboratory for Agriculture (LNNA), Embrapa Instrumentação, 13560-970, São Carlos, SP, Brazil.

出版信息

Talanta. 2022 Mar 1;239:123076. doi: 10.1016/j.talanta.2021.123076. Epub 2021 Nov 22.

Abstract

Mass testing for the diagnostics of COVID-19 has been hampered in many countries owing to the high cost of the methodologies to detect genetic material of SARS-CoV-2. In this paper, we report on a low-cost immunosensor capable of detecting the spike protein of SARS-CoV-2, including in samples of inactivated virus. Detection is performed with electrical impedance spectroscopy using an immunosensor that contains a monolayer film of carboxymethyl chitosan as matrix, coated with an active layer of antibodies specific to the spike protein. In addition to a low limit of detection of 0.179 fg/mL within an almost linear behavior from 10 g/mL to 10 g/mL, the immunosensor was highly selective. For the samples with the spike protein could be distinguished in multidimensional projection plots from samples with other biomarkers and analytes that could be interfering species for healthy and infected patients. The excellent analytical performance of the immunosensors was validated with the distinction between control samples and those containing inactivated SARS-CoV-2 at different concentrations. The mechanism behind the immunosensor performance is the specific antibody-protein interaction, as confirmed with the changes induced in C-H stretching and protein bands in polarization-modulated infrared reflection absorption spectra (PM-IRRAS). Because impedance spectroscopy measurements can be made with low-cost portable instruments, the immunosensor proposed here can be applied in point-of-care diagnostics for mass testing even in places with limited resources.

摘要

由于检测 SARS-CoV-2 遗传物质的方法成本高昂,许多国家的 COVID-19 诊断性大规模检测受到了阻碍。在本文中,我们报告了一种低成本的免疫传感器,能够检测 SARS-CoV-2 的刺突蛋白,包括已灭活病毒的样本。检测是通过使用包含羧甲基壳聚糖单层膜作为基质的免疫传感器进行的,该传感器涂有针对刺突蛋白的抗体的活性层。除了在 10 fg/mL 至 10 g/mL 之间几乎呈线性行为的情况下检测限低至 0.179 fg/mL 外,该免疫传感器还具有高度选择性。对于含有刺突蛋白的样本,可以通过多维投影图与含有其他生物标志物和分析物的样本区分开来,这些分析物可能是健康和感染患者的干扰物质。免疫传感器的出色分析性能通过区分不同浓度的对照样本和含有已灭活 SARS-CoV-2 的样本得到了验证。免疫传感器性能背后的机制是特定的抗体-蛋白相互作用,这一点通过在偏振调制红外反射吸收光谱(PM-IRRAS)中诱导的 C-H 伸缩和蛋白质带的变化得到了证实。由于阻抗光谱测量可以用低成本的便携式仪器进行,因此本文提出的免疫传感器可以应用于即时检测,用于大规模检测,即使在资源有限的地方也可以使用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/518b/8607795/289323a6d0d7/ga1_lrg.jpg

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