Abstract
Quantification of circulating microRNAs (miRNAs) or viral RNAs is of great significance because of their broad relevance to human health. Currently, quantitative reverse transcription polymerase chain reaction (qRT-PCR), as well as microarray and gene sequencing, are considered mainstream techniques for miRNA identification and quantitation and the gold standard for SARS-CoV2 detection in the COVID-19 pandemic. However, these laboratory techniques are challenged by the low levels and wide dynamic range (from aM to nM) of miRNAs in a physiological sample, as well as the difficulty in the implementation in point-of-care settings. Here, we describe a one-step label-free electrochemical sensing technique by assembling self-folded multi-stem DNA-redox probe structure on gold microelectrodes and introducing a reductant, tris(2-carboxyethyl) phosphine hydrochloride (TCEP), in the detection buffer solution to achieve ultrasensitive detection with a detection limit of 0.1 fM that can be further improved if needed.
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Acknowledgements
The microRNA work was supported by grants from the Department of Veterans Affairs (Merit Award: BX002526 to D. M.) and the National Institute of Health of Diabetes and Digestive and Kidney by the Grants RO1-DK-071594 and RO1-DK-064711 (to D. M.). Updated procedures and viral RNA analysis were developed under the financial support of NSF RAPID CHE-2034498.
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Ake, S., Kamila, S., Wang, G. (2023). Quantification of MicroRNAs or Viral RNAs with Microelectrode Sensors Enabled by Electrochemical Signal Amplification. In: Dalmay, T. (eds) MicroRNA Detection and Target Identification. Methods in Molecular Biology, vol 2630. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2982-6_9
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DOI: https://doi.org/10.1007/978-1-0716-2982-6_9
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