用于检测 SARS-CoV-2 的羧基-聚乙二醇-硫醇功能化金纳米粒子共轭物:检测工具和分析方法开发。

IF 2.2 4区 医学 Q3 BIOCHEMICAL RESEARCH METHODS Journal of virological methods Pub Date : 2024-09-03 DOI:10.1016/j.jviromet.2024.115028
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引用次数: 0

摘要

为了满足对 SARS-CoV-2 检测的需求,我们开发了羧基-PEG 12-硫醇功能化金纳米颗粒共轭物,用于针对 SARS-CoV-2 尖峰蛋白、伪 SARS-CoV-2 和真实 Beta SARS-CoV-2 病毒颗粒的快速床旁 (POC) 检测。这些共轭物利用金纳米粒子(AuNPs)作为信号转换器,与血管紧张素转换酶 2(ACE2)或 SARS-CoV-2 尖峰蛋白受体结合域(RBD)抗体交联作为生物受体,并显示出明显的从粉红色到蓝色的颜色变化。为了评估其 POC 可行性,我们将共轭物整合到了面罩和呼气分析仪中,成功检测到了气溶胶化的 SARS-CoV-2 抗原,呼气分析仪和面罩原型分别在 10 分钟和 30 分钟内产生了颜色变化。此外,我们还利用不同浓度的 SARS-CoV-2 尖峰蛋白进行了定量分析。两种共轭物都显示出蓝色强度与病毒浓度之间的线性关系,线性范围分别为 0.08-0.6ng/mL 和 0.04-0.5ng/mL。检测限和定量限也很低。它们具有特异性,即使在含有多种蛋白质(包括 SARS-CoV-1 尖峰蛋白)的复杂基质中,也只对 SARS-CoV-2 有反应。精确度测试的变异系数低于 2%,显示了其显著的可重复性。这项研究提出了一种快速、灵敏、特异的 SARS-CoV-2 POC 检测方法,为改进大流行病的应对和管理铺平了道路。
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Carboxy-PEG-thiol functionalized gold nanoparticle conjugates for the detection of SARS-CoV-2: Detection tools and analytical method development

Addressing the need for accessible SARS-CoV-2 testing, carboxy-PEG 12-thiol functionalized gold nanoparticles conjugates were developed for rapid point-of-care (POC) detection against SARS-CoV-2 spike protein, pseudo-SARS-CoV-2, and authentic Beta SARS-CoV-2 virus particles. These conjugates leverage gold nanoparticles (AuNPs) as signal transducers, cross-linked to either angiotensin-converting enzyme 2 (ACE2) or SARS-CoV-2 spike protein receptor-binding domain (RBD) antibodies as bioreceptors and showed a distinct color shift from pink to blue. To assess their POC feasibility, the conjugates were integrated into facemasks and breathalyzers, wherein aerosolized SARS-CoV-2 antigens were successfully detected, producing a color change within 10 and 30 minutes for the breathalyzer and facemask prototypes, respectively. Furthermore, we explored quantitative analysis using varying concentrations of SARS-CoV-2 spike protein. Both conjugates demonstrated a linear relationship between blue color intensity and virus concentration, with linear ranges of 0.08–0.6 ng/mL and 0.04–0.5 ng/mL, respectively. Low limits of detection and quantification were also achieved. They exhibited specificity, responding solely to SARS-CoV-2 even in complex matrices containing diverse proteins, including the SARS-CoV-1 spike protein. Precision tests yielded coefficient of variations below 2 %, showcasing their remarkable reproducibility. This work presents a promising approach for rapid, sensitive, and specific POC detection of SARS-CoV-2 paving the way for improved pandemic response and management.

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来源期刊
CiteScore
5.80
自引率
0.00%
发文量
209
审稿时长
41 days
期刊介绍: The Journal of Virological Methods focuses on original, high quality research papers that describe novel and comprehensively tested methods which enhance human, animal, plant, bacterial or environmental virology and prions research and discovery. The methods may include, but not limited to, the study of: Viral components and morphology- Virus isolation, propagation and development of viral vectors- Viral pathogenesis, oncogenesis, vaccines and antivirals- Virus replication, host-pathogen interactions and responses- Virus transmission, prevention, control and treatment- Viral metagenomics and virome- Virus ecology, adaption and evolution- Applied virology such as nanotechnology- Viral diagnosis with novelty and comprehensive evaluation. We seek articles, systematic reviews, meta-analyses and laboratory protocols that include comprehensive technical details with statistical confirmations that provide validations against current best practice, international standards or quality assurance programs and which advance knowledge in virology leading to improved medical, veterinary or agricultural practices and management.
期刊最新文献
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