CRISPR/Cas12-Driven Exponential Amplification Combined with a Lateral Flow Biosensor Enabling Rapid and Highly Sensitive DNA Detection

IF 9.1 1区化学 Q1 CHEMISTRY, ANALYTICAL

ACS Sensors Pub Date : 2025-09-12 DOI:10.1021/acssensors.5c00944

Mengqi Huang*, , , Kunte Shang, , , Luyi Ying, , , Yuqi Han, , , Ningxi Hong, , and , Yu-Feng Yao*,

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引用次数: 0

Abstract

Rapid and precise detection of specific DNA is valuable for biological research and clinical disease diagnosis. Clustered regularly interspaced short palindromic repeat (CRISPR) technology can enhance existing DNA testing, making DNA detection faster, more portable, and more accurate. This study presents a new Cas12-driven exponential amplification-based lateral flow biosensor (CADEX-LF) for rapid and highly sensitive DNA detection. CADEX-LF takes full advantage of the highly specific target-loading-dependent trans-cleavage activity of Cas12 and the extremely high efficiency of nicking endonuclease-mediated exponential amplification. The adoption of lateral flow readout enables CADEX-LF for point-of-care (POC) use without requiring complicated supplementary equipment. CADEX-LF was shown to achieve a detection sensitivity of 2 × 10^–15 M within 45 min of measurement time and displayed outstanding specificity with double-base resolution. Furthermore, CADEX-LF could identify herpes simplex virus 1 (HSV-1) DNA in tears of rabbits and clinical patients with HSV-1 keratitis, exhibiting its practical application potential in clinical diagnosis. The proposed CADEX-LF biosensor may have great promise for point-of-care disease diagnosis in resource-limited environments.

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CRISPR/ cas12驱动的指数扩增与横向流动生物传感器相结合，实现快速和高灵敏度的DNA检测。

快速、精确地检测特异性DNA对生物学研究和临床疾病诊断具有重要意义。聚类规则间隔短回文重复（CRISPR）技术可以增强现有的DNA检测，使DNA检测更快、更便携、更准确。本研究提出了一种新的基于cas12驱动指数扩增的横向流动生物传感器（CADEX-LF），用于快速和高灵敏度的DNA检测。CADEX-LF充分利用了Cas12高度特异性的靶向负载依赖的反式切割活性和极高效的切口内切酶介导的指数扩增。采用横向流量读数使CADEX-LF无需复杂的辅助设备即可用于护理点（POC）使用。CADEX-LF在45 min的测量时间内达到2 × 10-15 M的检测灵敏度，并且具有出色的双碱基分辨率特异性。此外，CADEX-LF还能在家兔和临床单纯疱疹病毒1型角膜炎患者的眼泪中识别单纯疱疹病毒1型（HSV-1） DNA，显示出其在临床诊断中的实际应用潜力。提出的CADEX-LF生物传感器可能在资源有限的环境中对即时疾病诊断有很大的希望。

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来源期刊

ACS Sensors Chemical Engineering-Bioengineering

CiteScore

14.50

自引率

3.40%

发文量

372

期刊介绍： ACS Sensors is a peer-reviewed research journal that focuses on the dissemination of new and original knowledge in the field of sensor science, particularly those that selectively sense chemical or biological species or processes. The journal covers a broad range of topics, including but not limited to biosensors, chemical sensors, gas sensors, intracellular sensors, single molecule sensors, cell chips, and microfluidic devices. It aims to publish articles that address conceptual advances in sensing technology applicable to various types of analytes or application papers that report on the use of existing sensing concepts in new ways or for new analytes.