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

IF 3.5 Q2 CHEMISTRY, ANALYTICAL
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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Abstract

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.

Abstract Image

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
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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CiteScore
2.30
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0.00%
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