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用于检测新型冠状病毒抗体的纤维素比色试纸条

Cellulose-Based Colorimetric Test Strips for SARS-CoV-2 Antibody Detection.

作者信息

Sousa Mariana P, Pereira Ana Cláudia, Correia Bárbara, do Carmo Anália, Matos Ana Miguel, Cruz Maria Teresa, Moreira Felismina T C

机构信息

BioMark/CEB/LABBELS-Centre of Biological Engineering, University of Minho, 4710-057 Braga, Portugal.

Epidemiology, Outcomes, Economics and Management in Oncology Group-Portuguese Oncology Institute of Porto Research Center (CI-IPOP), Porto Comprehensive Cancer Center (Porto.CCC) & RISE@CI-IPOP (Health Research Network), 4200-072 Porto, Portugal.

出版信息

Biosensors (Basel). 2025 Jun 17;15(6):390. doi: 10.3390/bios15060390.

Abstract

The COVID-19 pandemic highlighted the need for rapid, cost-effective tools to monitor transmission and immune response. We developed two novel paper-based colorimetric biosensors using glutaraldehyde as a protein dye-its first use in this context. Glutaraldehyde reacts with amino groups to generate a brown color, enabling detection of SARS-CoV-2 antibodies. Wathman filter paper was functionalized with (3-aminopropyl)triethoxysilane (APTES) to immobilize virus-like particles (VLPs) and nucleocapsid protein (N-protein) as biorecognition elements. Upon incubation with antibody-containing samples, glutaraldehyde enabled colorimetric detection using RGB analysis in ImageJ software. Both sensors showed a linear correlation between antibody concentration and RGB values in buffer and serum. The VLP sensor responded linearly within the range of 1.0-20 µg/mL (green coordinate) in 500-fold diluted serum and the N-protein sensor from 1.0-40 µg/mL (blue coordinate) in 250-fold diluted serum. Both sensors demonstrated good selectivity, with glucose causing up to 18% interference. These biosensors represent a paradigm shift, as they provide a sensitive, user-friendly, and cost-effective option for semi-quantitative serological analysis. Furthermore, their versatility goes beyond the detection of SARS-CoV-2 antibodies and suggests broader applicability for various molecular targets.

摘要

新冠疫情凸显了对用于监测病毒传播和免疫反应的快速、经济高效工具的需求。我们开发了两种新型的基于纸张的比色生物传感器,使用戊二醛作为蛋白质染料——这是其在此背景下的首次应用。戊二醛与氨基反应生成棕色,从而能够检测新冠病毒抗体。用(3-氨丙基)三乙氧基硅烷(APTES)对沃特曼滤纸进行功能化处理,以固定病毒样颗粒(VLP)和核衣壳蛋白(N蛋白)作为生物识别元件。在与含抗体的样品孵育后,戊二醛通过ImageJ软件中的RGB分析实现比色检测。两种传感器在缓冲液和血清中均显示出抗体浓度与RGB值之间的线性相关性。VLP传感器在500倍稀释血清中1.0 - 20 µg/mL(绿色坐标)范围内呈线性响应,N蛋白传感器在250倍稀释血清中1.0 - 40 µg/mL(蓝色坐标)范围内呈线性响应。两种传感器均表现出良好的选择性,葡萄糖造成的干扰高达18%。这些生物传感器代表了一种范式转变,因为它们为半定量血清学分析提供了一种灵敏、用户友好且经济高效的选择。此外,它们的多功能性不仅限于检测新冠病毒抗体,还表明对各种分子靶点具有更广泛的适用性。

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