Rapid and sensitive detection of nucleic acids has become crucial in various fields. However, most current nucleic acid detection methods can only be used in specific scenarios, such as RT-qPCR, which relies on fluorometer for signal readout, limiting its application at home or in the field due to its high price. In this paper, a universal nucleic acid detection platform combing CRISPR/Cas12a and strand displacement amplification (CRISPR-SDA) with multiple signal readout was established to adapt to different application scenarios. Nucleocapsid protein gene of SARS-CoV-2 (N gene) and hepatitis B virus (HBV) DNA were selected as model targets. The proposed strategy achieved the sensitivity of 53.1 fM, 0.15 pM, and 1 pM for N gene in fluorescence mode, personal glucose meter (PGM) mode and lateral flow assay (LFA) mode, respectively. It possessed the ability to differentiate single-base mismatch and the presence of salmon sperm DNA with a mass up to 10-fold of the targets did not significantly interfere with the assay signal. The general and modular design idea made CRISPR-SDA as simple as building blocks to construct nucleic acid sensing methods to meet different requirements by simply changing the SDA template and selecting suitable signal report probes, which was expected to find a breadth of applications in nucleic acids detection.

中文翻译：

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结合 CRISPR/Cas12a 和链置换扩增与多信号读出的通用核酸检测平台

快速、灵敏的核酸检测在各个领域变得至关重要。然而，目前大多数核酸检测方法只能在特定场景下使用，例如RT-qPCR，它依赖荧光计进行信号读出，由于价格昂贵，限制了其在家庭或现场的应用。本文建立了结合CRISPR/Cas12a和具有多种信号读出功能的链置换扩增（CRISPR-SDA）的通用核酸检测平台，以适应不同的应用场景。选择SARS-CoV-2的核衣壳蛋白基因（N基因）和乙型肝炎病毒（HBV）DNA作为模型目标。该策略在荧光模式、个人血糖仪（PGM）模式和侧流测定（LFA）模式下对N基因的灵敏度分别达到53.1 fM、0.15 pM和1 pM。它具有区分单碱基错配的能力，并且质量高达目标 10 倍的鲑鱼精子 DNA 的存在不会显着干扰检测信号。通用化、模块化的设计思想，使得CRISPR-SDA像搭积木一样简单，只需改变SDA模板和选择合适的信号报告探针，即可构建满足不同需求的核酸传感方法，有望在核酸领域得到广泛的应用检测。