通过将滚动圈扩增与基于聚(N-异丙基丙烯酰胺)的夹心型检测相结合,实现对 SARS-CoV-2 S1 蛋白的超灵敏检测

IF 5.6 1区 化学 Q1 CHEMISTRY, ANALYTICAL Talanta Pub Date : 2024-07-14 DOI:10.1016/j.talanta.2024.126572
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引用次数: 0

摘要

在过去几年中,由严重急性呼吸系统综合征冠状病毒 2(SARS-CoV-2)引起的 COVID-19 大流行因其高度传染性而严重威胁着全球公共卫生安全。开发一种快速灵敏的 SARS-CoV-2 检测方法仍然至关重要。在这项工作中,我们提出了一种基于聚(N-异丙基丙烯酰胺)(PNIPAM)的夹心型检测方法,可以在均相溶液中高效检测 SARS-CoV-2 S1 蛋白。首先,建立了一种直接夹心型检测方法,其线性范围为 0.2-2 μg/mL,检出限(LOD)为 0.11 μg/mL,可在约 1 小时内实现快速检测;此外,夹心型检测方法与滚动圆扩增(RCA)相结合,灵敏度提高了 5.9 × 104 倍,线性范围为 0.01 - 100 ng/mL,检出限(LOD)为 1.88 pg/mL。在未经处理的唾液中的平均回收率为 90 %-113.0 %,这表明所开发的方法具有在实际样品中应用的潜力。鉴于所开发方法的高选择性和高灵敏度,它在快速和早期检测SARS-CoV-2方面具有很大的潜力。
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Ultra-sensitive detection of SARS-CoV-2 S1 protein by coupling rolling circle amplification with poly(N-isopropylacrylamide)-based sandwich-type assay

In the past few years, the COVID-19 pandemic, caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) seriously threatens global public health security due to its high contagiousness. It remains of vital importance to develop a rapid and sensitive assay for SARS-CoV-2. In this work, we proposed a sandwich-type assay based on poly(N-isopropylacrylamide) (PNIPAM), allowing efficient detection of the SARS-CoV-2 S1 protein in the homogeneous solution. Firstly, a direct sandwich-type assay was established with a linear range of 0.2–2 μg/mL and a limit of detection (LOD) of 0.11 μg/mL, which could realize rapid detection in about 1 h. Furthermore, the sandwich-type assay coupled with rolling circle amplification (RCA) obtained an increase in sensitivity of 5.9 × 104 folds with a wide linear range of 0.01 − 100 ng/mL and a LOD of 1.88 pg/mL. The average recoveries in unpretreated saliva were 90 %–113.0 %, indicating the potential of the developed method for application in practical samples. Given the high selectivity and sensitivity of the developed method, it has a significant potential for rapid and early detection of SARS-CoV-2.

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来源期刊
Talanta
Talanta 化学-分析化学
CiteScore
12.30
自引率
4.90%
发文量
861
审稿时长
29 days
期刊介绍: Talanta provides a forum for the publication of original research papers, short communications, and critical reviews in all branches of pure and applied analytical chemistry. Papers are evaluated based on established guidelines, including the fundamental nature of the study, scientific novelty, substantial improvement or advantage over existing technology or methods, and demonstrated analytical applicability. Original research papers on fundamental studies, and on novel sensor and instrumentation developments, are encouraged. Novel or improved applications in areas such as clinical and biological chemistry, environmental analysis, geochemistry, materials science and engineering, and analytical platforms for omics development are welcome. Analytical performance of methods should be determined, including interference and matrix effects, and methods should be validated by comparison with a standard method, or analysis of a certified reference material. Simple spiking recoveries may not be sufficient. The developed method should especially comprise information on selectivity, sensitivity, detection limits, accuracy, and reliability. However, applying official validation or robustness studies to a routine method or technique does not necessarily constitute novelty. Proper statistical treatment of the data should be provided. Relevant literature should be cited, including related publications by the authors, and authors should discuss how their proposed methodology compares with previously reported methods.
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