What is it about?
We developed a 30-min microfluidic assay to detect SARS-CoV-2, the virus that causes COVID-19, using microfluidics technology. Our assay involves three stages including sample preparation, amplification, and CRISPR-Cas12 based nucleic acid detection. We achieve rapid and automated detection by the use of on-chip electric fields to extract and purify nucleic acids and to also control and accelerate CRISPR reaction kinetics to detect target nucleic acids, with each of these performed within 5 minutes. Our microfluidic device uses on-chip electric field control to generate electric field gradients that result in the co-location and preconcentration of target DNA, Cas12-gRNA and reporter molecules by ~1,000-fold and within a tiny ~100 pL volume, compared to conventional CRISPR reactions which are carried out in tubes. This automated preconcentration using ITP allows us to use much lower CRISPR reagents per test (about 100-fold lower) and also accelerates CRISPR-Cas12 enzymatic kinetics to enable rapid target detection.
Photo by CDC on Unsplash
Why is it important?
The COVID-19 pandemic has revealed major gaps in our current ability to respond to new pathogens. Rapid, accurate, and easily configurable molecular diagnostic tests are imperative to prevent global spread of new diseases. Early-stage screening and rapid identification of infected patients are also important to treat the infected in a timely manner. We believe field deployable and point-of-care testing methods offered by “lab on a chip” microfluidic technologies can address these requirements and radically change our approach to testing in the future.
Read the Original
This page is a summary of: Electric field-driven microfluidics for rapid CRISPR-based diagnostics and its application to detection of SARS-CoV-2, Proceedings of the National Academy of Sciences, November 2020, Proceedings of the National Academy of Sciences, DOI: 10.1073/pnas.2010254117.
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