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基于结构切换适体的纳米传感器用于水杨酸检测的研究进展。

Development of a structure-switching aptamer-based nanosensor for salicylic acid detection.

机构信息

Department of Plant Pathology and Microbiology, Iowa State University, Ames, IA, USA.

Department of Electrical and Computer Engineering, Iowa State University, Ames, IA, USA.

出版信息

Biosens Bioelectron. 2019 Sep 1;140:111342. doi: 10.1016/j.bios.2019.111342. Epub 2019 May 24.

Abstract

Salicylic acid (SA) is a phytohormone regulating immune responses against pathogens. SA and its derivatives can be found in diverse food products, medicines, cosmetics and preservatives. While salicylates have potential disease-preventative activity, they can also cause health problems to people who are hypersensitive. The current SA detection methods are costly, labor-intensive and require bulky instruments. In this study, a structure-switching aptamer-based nanopore thin film sensor was developed for cost-effective, rapid, sensitive and simple detection of SA in both buffer and plant extracts. SA is a challenging target for aptamer selection using conventional systemic evolution of ligands by exponential enrichment (SELEX) due to its small size and scarcity of reactive groups for immobilization. By immobilizing the SELEX library instead of SA and screening the library using a structure-switching SELEX approach, a high affinity SA aptamer was identified. The nanopore thin film sensor platform can detect as low as 0.1 μM SA. This is much better than the sensitivity of antibody-based detection method. This nanosensor also exhibited good selectivity among SA and its common metabolites and can detect SA in Arabidopsis and rice using only about 1 μl plant extracts within less than 30 min. The integration of SA aptamer and nanopore thin film sensor provides a promising solution for low-cost, rapid, sensitive on-site detection of SA.

摘要

水杨酸(SA)是一种调节植物免疫反应以抵御病原体的植物激素。SA 及其衍生物存在于各种食品、药品、化妆品和防腐剂中。虽然水杨酸盐具有潜在的疾病预防活性,但它们也会对过敏人群造成健康问题。目前的水杨酸检测方法成本高、劳动强度大且需要大型仪器。本研究开发了一种基于构象转换适体的纳米孔薄膜传感器,用于在缓冲液和植物提取物中进行经济、快速、灵敏和简单的水杨酸检测。由于 SA 体积小且用于固定化的反应基团稀缺,因此使用常规的指数富集配体系统进化(SELEX)技术来选择适体是具有挑战性的。通过固定化 SELEX 文库而不是 SA,并使用构象转换 SELEX 方法筛选文库,鉴定出了具有高亲和力的 SA 适体。纳米孔薄膜传感器平台可以检测低至 0.1 μM 的 SA。这比基于抗体的检测方法的灵敏度要好得多。该纳米传感器还表现出 SA 与其常见代谢物之间的良好选择性,并且仅使用约 1 μl 植物提取物在不到 30 分钟内即可在拟南芥和水稻中检测到 SA。SA 适体与纳米孔薄膜传感器的集成提供了一种有前途的解决方案,可用于低成本、快速、灵敏的现场检测 SA。

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