Electric-Fields-Enhanced CRISPR-Cas13a to detect SARS-CoV-2 in less than 20 min

Charles Blanluet, CentraleSupélec, Gif-sur-Yvette
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Department of Mechanical Engineering, Stanford University

Charles Blanluet

The COVID-19 pandemic has highlighted the importance of devices for rapid and sensitive detection of infectious diseases at the point of care (POC). A simplified, low-cost, and deployable assay for rapid and frequent tests for viral RNAs is needed to quickly assess and mitigate the progression of pandemics. We propose a low cost, deployable and sensitive detection microfluidic device which will use electric fields to enhance viral RNA assays. We will use a process called isotachophoresis (ITP) to increase the concentration of both viral material and CRISPR detection-complex between two ionic buffers. Consequently, for the same amount of viral RNA, the detection reaction is accelerated: CRISPR detection-complexes bind more often to viral RNA and cleave them, and this activates the enzyme complex. The activated complex thereafter cleaves quenched reporter molecules to release a fluorescence signal. My role will be to design and fabricate a microfluidic device to control the reaction, perform CRISPR assay experiments enhanced with ITP, and quantify the limits of sensitivity. The next step in the research will be to fully integrate the assay with viral RNA to DNA-reaction, amplification to multiply DNA and detection using ITP. The long-term goal is a portable electronic unit, powered and analyzed by a phone or a laptop that has the potential to revolutionize detection at the POC.

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