Accurate, rapid, cost-effective, and point-of-care pathogen identification is crucial for public health safety and encompasses disease prevention, food safety, and environmental governance. Despite the current prevalent detection methods that enable the inspection of various pathogens, certain limitations necessitate resolution. For instance, the plate culture method is time- and labor-intensive. Conversely, antibody- and large-equipment-based detection methods expedite the detection. However, their advancement has been constrained by the availability of antibodies and the inconvenience of deploying large equipment for point-of-care testing (POCT). The development of nucleic acid amplification technology can address the aforementioned issues to a certain extent. However, challenges such as intricate primer design and dependency on thermal cycling persist. Therefore, nucleic acid amplification-free biosensors have garnered significant attention. Biosensors are broadly categorized based on the following tools: nanomaterials-based nucleic acid amplification-free biosensors, clustered regularly interspaced short palindromic repeats/clustered regularly interspaced short palindromic repeat-associated nuclease (CRISPR/Cas)-based nucleic acid amplification-free biosensors, and nucleic acid amplification-free biosensors comprising nanomaterials and CRISPR/Cas. This review not only introduces biosensor performance and CRISPR/Cas applications, but also systematically elucidates the working principles and specific applications of nanomaterials and CRISPR/Cas in optimizing nucleic acid amplification-free detection systems, while judiciously categorizing them based on signal output modalities. Moreover, it discusses the challenges and prospects of nucleic acid amplification-free biosensors and provides valuable insights for further research.

中文翻译：

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下一代病原体检测：探索无核酸扩增生物传感器的力量

准确、快速、经济有效的即时病原体识别对于公共卫生安全至关重要，涵盖疾病预防、食品安全和环境治理。尽管目前流行的检测方法可以检查各种病原体，但仍需要解决某些局限性。例如，平板培养方法是时间和劳动力密集型的。相反，基于抗体和大型设备的检测方法可以加快检测速度。然而，它们的进步受到抗体可用性和部署大型设备进行即时检测（POCT）的不便的限制。核酸扩增技术的发展可以在一定程度上解决上述问题。然而，复杂的引物设计和对热循环的依赖等挑战仍然存在。因此，免核酸扩增生物传感器引起了人们的广泛关注。生物传感器根据以下工具大致分类：基于纳米材料的免核酸扩增生物传感器、基于簇状规则间隔短回文重复序列/簇状规则间隔短回文重复序列相关核酸酶（CRISPR/Cas）的免核酸扩增生物传感器、以及包含纳米材料和 CRISPR/Cas 的免核酸扩增生物传感器。本综述不仅介绍了生物传感器的性能和CRISPR/Cas的应用，还系统地阐述了纳米材料和CRISPR/Cas在优化核酸无扩增检测系统中的工作原理和具体应用，同时根据信号输出方式对其进行了明智的分类。此外，它还讨论了免核酸扩增生物传感器的挑战和前景，并为进一步的研究提供了宝贵的见解。