CRISPR-Responsive RCA-Based DNA Hydrogel Biosensing Platform with Customizable Signal Output for Rapid and Sensitive Nucleic Acid Detection

IF 6.7 1区化学 Q1 CHEMISTRY, ANALYTICAL

Analytical Chemistry Pub Date : 2024-09-25 DOI:10.1021/acs.analchem.4c0345010.1021/acs.analchem.4c03450

Yan Zhang, Weiwei Wang, Xinxi Zhou, Haonan Lin, Xiaotong Zhu, Yongliang Lou* and Laibao Zheng*,

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Abstract

Current nucleic acid-responsive DNA hydrogels face significant challenges, such as the requirement for high target concentrations, frequent redesigns, and increased costs, which limit their practical applications in biosensing. To address these issues, we developed a novel biosensing platform integrating a CRISPR/Cas12a system into an RCA-based DNA hydrogel. The hydrogel used in the platform could preencapsulate diverse signal molecules comprising GelRed, methylene blue, and gold nanoparticles, which were released upon Cas12a-mediated cleavage. This design enabled customizable signal output, including fluorescence, electrochemistry, and colorimetry, thereby ensuring the platform’s adaptability to various detection scenarios. Our platform was highly specific for methicillin-resistant Staphylococcus aureus, with a mecA gene detection limit of 10 copies/μL, and provided fast and accurate results within 2 h for clinical samples. Hence, based on these advantages, the proposed biosensing platform exhibits promising application prospects in the field of nucleic acid detection.

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基于 CRISPR 响应性 RCA 的 DNA 水凝胶生物传感平台，可定制信号输出，用于快速灵敏的核酸检测

目前的核酸响应 DNA 水凝胶面临着巨大的挑战，例如需要高目标浓度、频繁的重新设计以及成本增加，这些都限制了它们在生物传感领域的实际应用。为了解决这些问题，我们开发了一种新型生物传感平台，将 CRISPR/Cas12a 系统集成到基于 RCA 的 DNA 水凝胶中。该平台中使用的水凝胶可以预封装由 GelRed、亚甲基蓝和金纳米颗粒组成的多种信号分子，在 Cas12a 介导的裂解过程中释放出来。这种设计实现了可定制的信号输出，包括荧光、电化学和比色法，从而确保了平台对各种检测方案的适应性。我们的平台对耐甲氧西林金黄色葡萄球菌具有高度特异性，mecA 基因的检测限为 10 个拷贝/μL，并能在 2 小时内为临床样本提供快速准确的结果。因此，基于这些优点，该生物传感平台在核酸检测领域具有广阔的应用前景。

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来源期刊

Analytical Chemistry 化学-分析化学

CiteScore

12.10

自引率

12.20%

发文量

1949

审稿时长

1.4 months

期刊介绍： Analytical Chemistry, a peer-reviewed research journal, focuses on disseminating new and original knowledge across all branches of analytical chemistry. Fundamental articles may explore general principles of chemical measurement science and need not directly address existing or potential analytical methodology. They can be entirely theoretical or report experimental results. Contributions may cover various phases of analytical operations, including sampling, bioanalysis, electrochemistry, mass spectrometry, microscale and nanoscale systems, environmental analysis, separations, spectroscopy, chemical reactions and selectivity, instrumentation, imaging, surface analysis, and data processing. Papers discussing known analytical methods should present a significant, original application of the method, a notable improvement, or results on an important analyte.