School of Biomolecular Science and Engienering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, THAILAND

CRISPR-based diagnosis of infectious diseases: from the clinic to basics

Host: J. Molloy

I will discuss our development of CRISPR-based diagnostics for infectious diseases toward

point-of-care (POC) use outside of labs. We have developed a robust multiplexed CRISPR-

based detection using orthogonal CRISPR-associated nucleases with POC features to

simultaneously diagnose SARS-CoV-2 infection and pinpoint the causative SARS-CoV-2

variant of concern, and have implemented the CRISPR-based test technology for COVID-19

diagnosis with the approval of the FDA of Thailand. Beyond COVID-19, our team and

collaborators are applying CRISPR-based diagnostics to neglected fatal tropical diseases,

whose diagnosis is challenging yet critically important for timely treatment. One such

infectious disease is melioidosis, a bacterial infection with high fatality and incidence

rates in northeast Thailand. To reduce reliance on proprietary reagents and sustain use

of the technology in low- and middle-income countries, we locally produce a complex

recombinase polymerase isothermal amplification (RPA) reaction typically used in

conjunction with CRISPR-based detection and demonstrate its equivalent efficiency to

commercial counterparts. The ability to produce RPA in-house enables us to uncover the

previously unknown core feature of RPA—that the reaction is catalyzed within multiphasic

condensates—and to engineer its bottleneck components, further improving the reaction

for multiplexed diagnosis.


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