CRISPR-Cas12a-mediated label-free electrochemical aptamer-based sensor for SARS-CoV-2 antigen detection.
CRISPR-Cas12a-mediated label-free electrochemical aptamer-based sensor for SARS-CoV-2 antigen detection.
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DOI：
10.1016/j.bioelechem.2022.108105
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发表时间：
2022-08
期刊：
影响因子：
--
通讯作者：
Zhang J
中科院分区：
文献类型：
--
作者：
Liu N;Liu R;Zhang J
血清学抗原检测已成为COVID-19的重要诊断模式，但通常存在潜在的交叉反应。为了解决这一局限性，我们在此报告了一种无标记的基于电化学适体的传感器，用于通过整合基于适体的特异性识别与CRISPR-Cas 12 a介导的信号放大来检测SARS-CoV-2抗原。传感原理基于抗原和预组装的Cas 12 a-crRNA复合物与抗原特异性适体的竞争性结合，导致Cas 12 a的附带切割活性的变化。为了进一步产生电化学信号，通过在金电极上原位滚环扩增来制造DNA结构，该金电极用作Cas 12 a的新型基底。在基于Cas 12 a的侧枝DNA切割后，DNA结构被降解，导致可以通过光谱测量的阻抗显著降低。以SARS-CoV-2核衣壳抗原为模型，基于CRISPR-Cas 12 a的电化学传感器（CRISPR-E）对核衣壳抗原的定量检测表现出优异的分析性能。由于体外选择可以获得对许多SARS-CoV-2抗原具有选择性的适体，因此所提出的策略可以显着扩展这种强大的CRISPR-E系统，用于定量监测各种COVID-19生物标志物。
Serological antigen testing has emerged as an important diagnostic paradigm in COVID-19, but often suffers from potential cross-reactivity. To address this limitation, we herein report a label-free electrochemical aptamer-based sensor for the detection of SARS-CoV-2 antigen by integrating aptamer-based specific recognition with CRISPR-Cas12a-mediated signal amplification. The sensing principle is based on the competitive binding of antigen and the preassembled Cas12a-crRNA complex to the antigen-specific aptamer, resulting in a change in the collateral cleavage activity of Cas12a. To further generate an electrochemical signal, a DNA architecture was fabricated by in situ rolling circle amplification on a gold electrode, which serves as a novel substrate for Cas12a. Upon Cas12a-based collateral DNA cleavage, the DNA architecture was degraded, leading to a significant decrease in impedance that can be measured spectroscopically. Using SARS-CoV-2 nucleocapsid antigen as the model, the proposed CRISPR-Cas12a-based electrochemical sensor (CRISPR-E) showed excellent analytical performance for the quantitative detection of nucleocapsid antigen. Since in vitro selection can obtain aptamers selective for many SARS-CoV-2 antigens, the proposed strategy can expand this powerful CRISPR-E system significantly for quantitative monitoring of a wide range of COVID-19 biomarkers.
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DOI：
10.1016/j.cej.2021.131433
发表时间：
2021-12-01
期刊：
Chemical engineering journal (Lausanne, Switzerland : 1996)
影响因子：
--
作者：
Muñoz J;Pumera M
通讯作者：
Pumera M
DOI：
10.1016/j.cej.2021.132966
发表时间：
2022-02-15
期刊：
Chemical engineering journal (Lausanne, Switzerland : 1996)
影响因子：
--
作者：
Kumar N;Shetti NP;Jagannath S;Aminabhavi TM
通讯作者：
Aminabhavi TM
影响因子：
16.6
作者：
Ding X;Yin K;Li Z;Lalla RV;Ballesteros E;Sfeir MM;Liu C
通讯作者：
Liu C
ZIF-8 辅助的 NaYF4:Yb,Tm@ZnO 转换器，具有核酸外切酶 III 驱动的 DNA 步行器，用于近红外光响应生物传感器
影响因子：
7.4
作者：
Lv, Shuzhen;Zhang, Kangyao;Tang, Dianping
通讯作者：
Tang, Dianping
影响因子：
16.6
作者：
Dai, Yifan;Somoza, Rodrigo A.;Liu, Chung Chiun
通讯作者：
Liu, Chung Chiun