Sample-in-answer-out centrifugal microfluidic chip reaction biosensor powered by Thermus thermophilus Argonaute (TtAgo) for rapid, highly sensitive and multiplexed molecular diagnostics of foodborne bacterial pathogens

• 全文链接

由Thermus thermophilus Argonaute (TtAgo)驱动的样品-应答-输出离心式微流控芯片反应生物传感器，用于快速、高灵敏度和多重分子诊断食源性细菌病原体

关键词：

来源：

CHEMICAL ENGINEERING JOURNAL

全文链接：

//agri.nais.net.cn/topic/downloadFile/b56649c8-45d4-4392-8f36-4d589745e6ab

来源地址：

https://linkinghub.elsevier.com/retrieve/pii/S1385894724049234

资源所属：

农牧业信息化专题

类型：

学术文献

语种：

英语

原文发布日期：

2024-07-01

摘要：

Foodborne pathogens endanger public health and rapid, sensitive and accurate detection of them is vital. Argonaute stands in the frontline as the next generation nucleic acid detection tool, bearing a few comparable advantages. Centrifugal microfluidic chips (CMCs) use centrifugal force to achieve liquid flowing, mixing and reaction, eliminating complicated designs of valves and pumps. In this study, for the first time, we devised a Thermus thermophilus Argonaute (TtAgo)-powered centrifugal microfluidic chip reaction biosensor for Sample-inAnswer-out detection of Pathogen S. aureus (termed as ASAP) with ultrahigh rapidity and sensitivity in a one-pot fashion. The samples subjected to testing were mixed with a home-made nucleic acid fast extraction reagent and then the mixture was injected into the sample cell of CMC, which was centrifuged down into the reaction cell. The reaction cell was preloaded with both LAMP (Loop-mediated isothermal amplification) and TtAgo systems. As such, the species-specific nuc genes were amplified by LAMP, while the TtAgo performed site-specific cleavage to output fluorescent (FL) signals. The sample-to-answer time was 17 min, and the limit of detection (LOD) reached 1 CFU/mL. ASAP was able to perform simultaneous multiplexed detection and up to 16 samples can be detected at the same time. ASAP reduced reagent consumption and minimized the influence of carry-over and cross-contamination. ASAP was capable of detecting S. aureus in foods, but also detecting physiological samples from infect, holding great promise for practical applications. Overall, our work has enriched the Argonautepowered biosensing and CMC biosensor technology by providing a conceptually novel bacterial detection platform.