利用分子信标探针对细胞内 mRNA 进行微流控测量,实现医护点辐射分诊

IF 3.5 Q2 CHEMISTRY, ANALYTICAL Sensors & diagnostics Pub Date : 2024-07-02 DOI:10.1039/D4SD00079J
Xin Meng, Kechun Wen, Jingyang Zhao, Yaru Han, Shanaz A. Ghandhi, Salan P. Kaur, David J. Brenner, Helen C. Turner, Sally A. Amundson and Qiao Lin
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引用次数: 0

摘要

在大规模辐照事件中,根据接受的辐射剂量对潜在受害者进行分流的能力至关重要。这可以通过辐射诱导的生物变化来评估。辐射反应 mRNA 是一类生物标志物,已通过 RT-qPCR 等方法探索了其剂量依赖性。然而,这些方法在护理点设备上的实施具有挑战性。我们设计并使用分子信标作为探针,在微流控装置中测量辐射诱导的细胞内 mRNA 变化,以确定辐射剂量。在我们的实验中,用特异性 BAX mRNA 分子信标标记的固定 TK6 细胞表现出剂量依赖性荧光,其方式与 RT-qPCR 分析一致。这一概念验证可随时扩展到任何基于 RNA 的检测,以提供床旁直接测量。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Microfluidic measurement of intracellular mRNA with a molecular beacon probe towards point-of-care radiation triage†

In large-scale radiation exposure events, the ability to triage potential victims by the received radiation dosage is crucial. This can be evaluated by radiation-induced biological changes. Radiation-responsive mRNA is a class of biomarkers that has been explored for dose-dependency with methods such as RT-qPCR. However, these methods are challenging to implement for point-of-care devices. We have designed and used molecular beacons as probes for the measurement of radiation-induced changes of intracellular mRNA in a microfluidic device towards determining radiation dosage. Our experiments, in which fixed TK6 cells labeled with a molecular beacon specific to BAX mRNA exhibited dose-dependent fluorescence in a manner consistent with RT-qPCR analysis, demonstrate that such intracellular molecular probes can potentially be used in point-of-care radiation biodosimetry. This proof of concept could readily be extended to any RNA-based test to provide direct measurements at the bedside.

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