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基于液滴的DNA模板纳米簇测量——迈向即时检测应用

Droplet-Based Measurements of DNA-Templated Nanoclusters-Towards Point-of-Care Applications.

作者信息

Kluitmann Jonas, Di Fiore Stefano, Nölke Greta, Drese Klaus Stefan

机构信息

Institute of Sensor and Actuator Technology, Coburg University of Applied Sciences and Arts, Am Hofbräuhaus 1b, 96450 Coburg, Germany.

Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Forckenbeckstr. 6, 52074 Aachen, Germany.

出版信息

Biosensors (Basel). 2025 Jul 1;15(7):417. doi: 10.3390/bios15070417.

Abstract

In this work, we investigate the fundamental usability of fluorescent DNA-templated silver nanoclusters (DNA-AgNCs) as sensors for Point-of Care-Testing (PoCT) applications. We developed a microfluidic platform for the generation of droplets containing DNA-AgNCs in defined, different chemical environments. The droplets are read out fluorescently at two different emission wavelengths. For the pre-evaluation for the usage of biologically relevant matrices with DNA-AgNCs, the response of two different DNA-AgNCs to a variation in pH and sodium chloride concentration was acquired. Our compact and simple setup can detect DNA-AgNCs well below 100 nM and allows the characterization of the fluorescence response of DNA-based biohybrid nanosensors to changes in the chemical environment within short measurement times. The model DNA-AgNCs remain fluorescent throughout the physiologically relevant chloride concentrations and up to 150 mM. Upon shifts in pH, the DNA-AgNCs showed a complex fluorescence intensity response. The model DNA-AgNCs differ strongly in their response characteristics to the applied changes in their environments. With our work, we show the feasibility of the use of DNA-AgNCs as sensors in a simple microfluidic setup that can be used as a building block for PoCT applications while highlighting challenges in their adaption for use with biologically relevant matrices.

摘要

在这项工作中,我们研究了荧光DNA模板化银纳米簇(DNA-AgNCs)作为即时检测(PoCT)应用传感器的基本可用性。我们开发了一种微流控平台,用于在特定的不同化学环境中生成含有DNA-AgNCs的液滴。这些液滴在两个不同的发射波长下进行荧光读取。为了对DNA-AgNCs在生物相关基质中的使用进行预评估,我们获取了两种不同的DNA-AgNCs对pH值和氯化钠浓度变化的响应。我们紧凑且简单的装置能够检测浓度远低于100 nM的DNA-AgNCs,并能在短测量时间内对基于DNA的生物杂交纳米传感器对化学环境变化的荧光响应进行表征。在整个生理相关的氯化物浓度范围内以及高达150 mM时,模型DNA-AgNCs都保持荧光。随着pH值的变化,DNA-AgNCs呈现出复杂的荧光强度响应。模型DNA-AgNCs对所施加的环境变化的响应特性差异很大。通过我们的工作,我们展示了在简单微流控装置中使用DNA-AgNCs作为传感器的可行性,该装置可作为PoCT应用的构建模块,同时突出了其在适应生物相关基质使用时所面临的挑战。

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