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

使用分子信标探针进行细胞内 mRNA 微流体测量以实现即时放射分类

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作者:Xin Meng, Kechun Wen, Jingyang Zhao, Yaru Han, Shanaz A Ghandhi, Salan P Kaur, David J Brenner, Helen C Turner, Sally A Amundson, Qiao Lin

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.

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