Building Novel Point-of-Care Diagnostics Through CRISPR Diagnostics

Clinical Center

Investigators

Abstract

We developed a bioinformatic pipeline to nominate RPA amplicons and CRISPR guide RNAs, with in-silico inclusivity/coverage and cross-reactivity screening against public genomes. Using this pipeline, we built and optimized multiplex CRISPR-Cas12a/RPA assays for Mycobacterium tuberculosis, M. avium complex, and M. abscessus, and for priority carbapenemase determinants (KPC, NDM, VIM, OXA-48-like, IMP). We demonstrated functional performance on routine microbiology specimen types—directly from colony material on agar, from standard culture media (e.g., MGIT broth), and from positive blood culture bottles—establishing feasibility on standard laboratory workflows. These results support rapid species and resistance marker detection suitable for decentralized settings and stewardship-oriented decisions.