Clinically applicable assay of cardiac troponin I by CRISPR/Cas12a-amplified single-particle collision electrochemistry

IF 8 1区化学 Q1 CHEMISTRY, ANALYTICAL

Sensors and Actuators B: Chemical Pub Date : 2024-03-27 DOI:10.1016/j.snb.2024.135708

Jinrong Liu , Long Mi , Jie Yang , Hui Qian , Heng Liu , Wei Wen , Xiuhua Zhang , Zhen Wu , Shengfu Wang

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Abstract

Owing to its rapid response and ultra-high sensitivity at single-entity level, single-particle collision electrochemistry (SPCE) shows great application prospects in sensitive detection of biomolecules. However, its widespread use in actual samples is constrained by the poor signal conversion efficiency and heavy interference from complicated samples. Here, a new SPCE biosensor for clinical analysis of cardiac troponin I (cTnI) was developed based on the effective signal amplification of CRISPR/Cas12a as well as the good separation and enrichment capabilities of magnetic nanobeads (MBs). The trans-cleavage capability of CRISPR/Cas12a can only be triggered in the presence of target cTnI to effectively cleave ssDNA₂ on the surface of Au@Pt nanoparticles (Au@Pt NPs) within 30 min. Each cTnI molecule could cause the release of 4.68×10⁴ Au@Pt NPs by utilizing the superior cleaving activity of CRISPR/Cas12a, significantly amplifying the signal and improving the sensitivity of analysis. The developed SPCE biosensor for cTnI had a detection limit as low as 0.35 fg/mL, which was 2–6 orders in magnitude less than existing cTnI detection techniques. Additionally, MBs allow the biosensor to detect cTnI in patient samples with excellent sensitivity and specificity by directly sampling from the complex matrix.

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通过 CRISPR/Cas12a 扩增单粒子碰撞电化学法测定临床适用的心肌肌钙蛋白 I

单粒子碰撞电化学（SPCE）因其快速响应和单实体水平的超高灵敏度，在生物大分子的灵敏检测方面具有广阔的应用前景。然而，在实际样品中的广泛应用却受到信号转换效率低、复杂样品干扰大等因素的制约。本文基于 CRISPR/Cas12a 的有效信号放大和磁性纳米eads（MBs）良好的分离和富集能力，开发了一种用于临床分析心肌肌钙蛋白 I（cTnI）的新型 SPCE 生物传感器。CRISPR/Cas12a的反式裂解能力只有在目标cTnI存在的情况下才能被触发，从而在30分钟内有效地裂解金@铂纳米颗粒（Au@Pt NPs）表面的ssDNA2。利用 CRISPR/Cas12a 卓越的裂解活性，每个 cTnI 分子可导致 4.68×104 个 Au@Pt NPs 释放，显著放大了信号并提高了分析灵敏度。所开发的 cTnI SPCE 生物传感器的检测限低至 0.35 fg/mL，比现有的 cTnI 检测技术低 2-6 个数量级。此外，甲基溴允许生物传感器直接从复杂基质中采样，从而检测患者样本中的 cTnI，具有极佳的灵敏度和特异性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Sensors and Actuators B: Chemical 工程技术-电化学

CiteScore

14.60

自引率

11.90%

发文量

1776

审稿时长

3.2 months

期刊介绍： Sensors & Actuators, B: Chemical is an international journal focused on the research and development of chemical transducers. It covers chemical sensors and biosensors, chemical actuators, and analytical microsystems. The journal is interdisciplinary, aiming to publish original works showcasing substantial advancements beyond the current state of the art in these fields, with practical applicability to solving meaningful analytical problems. Review articles are accepted by invitation from an Editor of the journal.