Exonuclease III assisted exponential amplification reaction (EXPAR) for specific miRNA-155 analysis during post-anesthetic nursing

IF 2.5 4区 化学 Q3 CHEMISTRY, ANALYTICAL Journal of Analytical Science and Technology Pub Date : 2024-06-24 DOI:10.1186/s40543-024-00454-0
Xuejun Wu, Shaolan Zou, Jingshen Dai
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Abstract

The persistent obstacle in precise and sensitive identification of microRNAs (miRNAs) pertains to the advancement of expeditious and effective isothermal amplification methodologies suitable for point-of-care environments and monitoring the cancer prognosis in patients receiving post-anesthetic nursing. The exponential amplification reaction (EXPAR) has attracted considerable interest due to its simplicity and ability to rapidly amplify signals. The practical application of the EXPAR is, nevertheless, severely hampered by the inability to differentiate closely related homologous sequences and to modify the designed templates to suit other targets. A loop-stem template for the EXPAR system was developed in this study to facilitate specific target recognition with the aid of exonuclease III (Exo III). This innovation effectively eliminated non-specific hybridization that could occur between the template and interfering sequences, thereby ensuring minimal background amplification of EXPAR. By modulating Exo III-based target recycling, EXPAR based chain amplification and G4/hemin based color reaction, this method facilitated the precise and sensitive examination of miRNA-155, yielding acceptable yields and a minimal detection limit of 0.43 fM. The approach expedites simple and expeditious molecular diagnostic applications involving short nucleic acids and offers an innovative method for enhancing the selectivity of EXPAR-based techniques, providing a robust tool for monitoring the expression level from patients receiving post-anesthetic nursing and guiding the treatment strategy.
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外切酶 III 辅助指数扩增反应 (EXPAR) 用于麻醉后护理期间 miRNA-155 的特异性分析
在精确、灵敏地鉴定微RNA(miRNA)方面一直存在的障碍是,如何开发快速、有效的等温扩增方法,以适用于护理点环境和监测接受麻醉后护理的患者的癌症预后。指数扩增反应(EXPAR)因其简便性和快速扩增信号的能力而备受关注。然而,由于无法区分密切相关的同源序列,也无法修改所设计的模板以适应其他目标,EXPAR 的实际应用受到严重阻碍。本研究为 EXPAR 系统开发了一种环茎模板,以便借助外切酶 III(Exo III)进行特异性目标识别。这一创新有效消除了模板与干扰序列之间可能发生的非特异性杂交,从而确保 EXPAR 的背景扩增最小化。通过调节基于 Exo III 的目标回收、基于 EXPAR 的链扩增和基于 G4/hemin 的颜色反应,该方法有助于精确灵敏地检测 miRNA-155,产量可接受,最低检测限为 0.43 fM。该方法加快了涉及短核酸的简单快捷的分子诊断应用,为提高基于 EXPAR 的技术的选择性提供了一种创新方法,为监测接受麻醉后护理的患者的表达水平和指导治疗策略提供了一种强有力的工具。
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来源期刊
Journal of Analytical Science and Technology
Journal of Analytical Science and Technology Environmental Science-General Environmental Science
CiteScore
4.00
自引率
4.20%
发文量
39
审稿时长
13 weeks
期刊介绍: The Journal of Analytical Science and Technology (JAST) is a fully open access peer-reviewed scientific journal published under the brand SpringerOpen. JAST was launched by Korea Basic Science Institute in 2010. JAST publishes original research and review articles on all aspects of analytical principles, techniques, methods, procedures, and equipment. JAST’s vision is to be an internationally influential and widely read analytical science journal. Our mission is to inform and stimulate researchers to make significant professional achievements in science. We aim to provide scientists, researchers, and students worldwide with unlimited access to the latest advances of the analytical sciences.
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