据估计，超过 50% 的人类癌症是由 p53 基因突变引起的。 p53基因的早期灵敏和准确检测对于癌症的早期诊断非常重要。然而，传统的检测技术往往反应条件严格，或者灵敏度不理想，因此我们需要开发一种新的策略来精确检测p53基因，具有智能设计性，在温和的反应条件下进行多重信号放大。在本研究中，CRISPR/Cas系统用于熵驱动催化 (EDC) 和杂交链式反应 (CHA) 双信号放大传感策略。两种反应的产物都可以有效地、单独地激活CRISPR/Cas12a，从而极大地放大荧光信号。该方法在p53检测中具有良好的线性，浓度范围为0.1 fM至0.5 pM，超低检测限为0.096 fM。它还在血清中表现出良好的性能，为早期疾病检测提供了潜力。设计的双扩增动态DNA网络系统表现出用于p53基因识别的超灵敏荧光生物传感。该方法操作简单，实验只需要一种缓冲液，同时显示出智能的可设计性，可用于广泛的标记。因此，我们相信目前的工作将为构建和开发疾病敏感荧光生物传感器提供潜在的工具。版权所有 © 2024 Elsevier B.V. 保留所有权利。

It is estimated that over 50 % of human cancers are caused by mutations in the p53 gene. Early sensitive and accurate detection of the p53 gene is important for diagnosis of cancers in the early stage. However, conventional detection techniques often suffer from strict reaction conditions, or unsatisfied sensitivity, so we need to develop a new strategy for accurate detection of p53 gene with smart designability, multiple signal amplification in mild reaction conditions.In this study, CRISPR/Cas system is exploited in entropy-driven catalysis (EDC) and hybridization chain reaction (CHA) dual signal amplification sensing strategies. The products of both reactions can efficiently and separately activate CRISPR/Cas12a which greatly amplifies the fluorescent signal. The method has good linearity in p53 detection with the concentration ranged from 0.1 fM to 0.5 pM with ultra-low detection limit of 0.096 fM. It also showed good performance in serum, offering potentials for early disease detection.The designed dual amplification dynamic DNA network system exhibits an ultra-sensitive fluorescence biosensing for p53 gene identification. The method is simple to operate and requires only one buffer for the experiment, and meanwhile shows smart designability which could be used for a wide range of markers. Thus, we believe the present work will provide a potential tool for the construction and development of sensitive fluorescent biosensors for diseases.Copyright © 2024 Elsevier B.V. All rights reserved.