Xingyu Ye, Haoyang Wu, Jinghan Liu, Jiayi Xiang, Yan Feng, Qian Liu
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引用次数: 0
摘要
针对特定核酸序列的 CRISPR/Cas 和 Argonaute（Ago）蛋白可用作诊断工具。尽管具有高特异性和高效性，但要实现灵敏检测，往往需要进行预扩增步骤，包括打开盖子和多步操作，这可能会增加污染风险，并证明不适用于护理点检测。因此，人们开发了各种单锅检测策略，只需一次操作即可完成前置扩增和传感。我们概述了使用 Cas 和 Ago 蛋白进行一次性检测所面临的挑战，介绍了几种主要的实施策略，并讨论了未来的前景。这篇综述提供了对这一重要领域的全面见解，并探讨了对检测方法的潜在改进，这将有益于人类健康。
One-pot diagnostic methods based on CRISPR/Cas and Argonaute nucleases: strategies and perspectives.
CRISPR/Cas and Argonaute (Ago) proteins, which target specific nucleic acid sequences, can be applied as diagnostic tools. Despite high specificity and efficiency, achieving sensitive detection often necessitates a preamplification step that involves opening the lid and multistep operation, which may elevate the risk of contamination and prove inadequate for point-of-care testing. Hence, various one-pot detection strategies have been developed that enable preamplification and sensing in a single operation. We outline the challenges of one-pot detection with Cas and Ago proteins, present several main implementation strategies, and discuss future prospects. This review offers comprehensive insights into this vital field and explores potential improvements to detection methods that will be beneficial for human health.
期刊介绍：
Trends in Biotechnology publishes reviews and perspectives on the applied biological sciences, focusing on useful science applied to, derived from, or inspired by living systems.
The major themes that TIBTECH is interested in include:
Bioprocessing (biochemical engineering, applied enzymology, industrial biotechnology, biofuels, metabolic engineering)
Omics (genome editing, single-cell technologies, bioinformatics, synthetic biology)
Materials and devices (bionanotechnology, biomaterials, diagnostics/imaging/detection, soft robotics, biosensors/bioelectronics)
Therapeutics (biofabrication, stem cells, tissue engineering and regenerative medicine, antibodies and other protein drugs, drug delivery)
Agroenvironment (environmental engineering, bioremediation, genetically modified crops, sustainable development).
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