Nanopore-assisted ELISA for ultrasensitive, portable, and on-site detection of ricin.

IF 5.6 1区化学 Q1 CHEMISTRY, ANALYTICAL Talanta Pub Date : 2025-02-01 Epub Date: 2024-10-31 DOI:10.1016/j.talanta.2024.127136

Jianing Chen, Zhuoqun Su, Wenrui Li, Ziye Pei, Di Wu, Lin Li, Yongning Wu, Guoliang Li

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Abstract

Effective detection technologies in food safety with the merits of portable and on-site detection potential are always in pressing demand. Herein, we developed a nanopore-assisted Enzyme-linked immunosorbent assay (NELISA) platform, which innovatively introduced hairpin DNA (HP DNA) probes as reaction substrates. This innovation of substrates effectively avoided the inherent limitations of colorimetric signals (i.e., low sensitivity and inaccurate results) and greatly improved the sensitivity and accuracy of NELISA platform. The alkaline phosphatase (ALP)-modified detection antibody (ALP-Ab2) can specifically bind to ricin and hydrolyze the phosphate groups modified on the HP DNA probes. Nanopore recordings demonstrated that two states of probes produced highly distinguishable nanopore events, enabling the qualitative and quantitative detection of ricin. This NELISA platform fully combined the specificity of ELISA with the ultra-sensitivity, and unique single-molecule fingerprint recognition of nanopore, showing a great on-site detection potential. This method achieved the ultrasensitive and reliable detection of ricin down to 2.46 fg/mL, which enhanced the detection sensitivity by at least 10⁶-fold compared to traditional ELISA. Furthermore, the proposed method was capable of accurately detecting ricin in real food samples with satisfactory recoveries.

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用于超灵敏、便携和现场检测蓖麻毒素的纳米孔辅助酶联免疫吸附试验。

食品安全领域一直迫切需要具有便携性和现场检测潜力的有效检测技术。在此，我们开发了一种纳米孔辅助酶联免疫吸附测定（NELISA）平台，创新性地引入了发夹式 DNA（HP DNA）探针作为反应底物。这种底物的创新有效避免了比色信号的固有局限性（即灵敏度低、结果不准确），大大提高了 NELISA 平台的灵敏度和准确性。碱性磷酸酶（ALP）修饰的检测抗体（ALP-Ab2）能与蓖麻毒素特异性结合，并水解 HP DNA 探针上修饰的磷酸基团。纳米孔记录表明，探针的两种状态会产生高度可区分的纳米孔事件，从而实现对蓖麻毒素的定性和定量检测。这种 NELISA 平台充分结合了 ELISA 的特异性和纳米孔的超灵敏性以及独特的单分子指纹识别功能，显示出巨大的现场检测潜力。该方法实现了对低至2.46 fg/mL的蓖麻毒素的超灵敏可靠检测，与传统的ELISA相比，检测灵敏度至少提高了106倍。此外，该方法还能准确检测真实食品样品中的蓖麻毒素，回收率令人满意。

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来源期刊

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.