School of Food Science and Engineering, Shaanxi University of Science and Technology, Xi'an 710021, China.
Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, 19 Chlorine Gardens, Belfast BT9 5DL, United Kingdom.
J Agric Food Chem. 2024 Jan 17;72(2):1354-1360. doi: 10.1021/acs.jafc.3c07881. Epub 2024 Jan 4.
Argonaute (Ago) emerged as a novel endonuclease for the nucleic acid test recently. However, the input of exogenous guide DNA (gDNA) to activate Ago has reduced its flexibility. In this work, an enzyme-assisted ndogenous DNA eneration-mediated go for the target etection strategy, termed EGG-PAD, was proposed. With the aid of EcoR Ι, the target double-strand DNA was cut, producing a phosphate group at the 5' end, functioning as gDNA to activate Ago for nucleic acid detection. The applicability of this assay was tested in the detection of, a bacterium causing the spoilage of fruit juice, showing excellent sensitivity and specificity, ascribed to the "duplex amplification and triple insurance" mechanism. Moreover, EGG-PAD exhibited superior versatility in the identification of common foodborne pathogens. This powerful platform could also be an on-site test tool for detecting nucleic acid-containing organisms such as tumor cell, pathogen, and virus in the future.
Argonaute（AGO）最近作为一种新型核酸内切酶出现在核酸检测中。然而，外源性引导 DNA（gDNA）的输入激活 AGO 降低了其灵活性。在这项工作中，提出了一种酶辅助的内源性 DNA 生成介导的靶向检测策略，称为 EGG-PAD。借助 EcoR Ι，靶双链 DNA 被切割，在 5' 端产生一个磷酸基团，作为 gDNA 激活 AGO 进行核酸检测。该测定方法在检测导致果汁变质的细菌中的应用进行了测试，表现出优异的灵敏度和特异性，归因于“双链扩增和三重保险”机制。此外，EGG-PAD 在鉴定常见食源性病原体方面表现出更高的通用性。这个强大的平台将来也可能成为现场检测工具，用于检测肿瘤细胞、病原体和病毒等含有核酸的生物。
