J.B. Scott, C.H. Monk, K.J. Zwezdaryk
Tulane University,
United States
Keywords: CRISPR-Cas12a, Padlock probe, Isothermal amplification, Molecular diagnostics, Point-of-care testing
Summary:
This technology introduces an innovative diagnostic platform that combines CRISPR-Cas12a with padlock probe-based Rolling Circle Amplification (RCA) for the detection of specific nucleic acid targets in biological samples. Unlike traditional molecular diagnostics that rely on PCR pre-amplification steps, this method employs isothermal amplification techniques that occur at near room temperature in as little as 20-30 minutes. The system allows for highly specific targeting of genomic regions, thereby increasing copy number and enhancing assay sensitivity and specificity. Initially developed for the detection of Human Cytomegalovirus (CMV), this technology has broad applications across molecular genomic diagnostics. The method significantly reduces the need for sophisticated equipment and complex sample preparation, making it ideal for point-of-care settings and resource-limited environments. This innovation addresses critical healthcare needs by enabling rapid, sensitive, and specific diagnostics with minimal instrumentation requirements. A spin-out company, Cleaved Diagnostics, has been formed to further develop and commercialize this platform, which has the potential to revolutionize molecular diagnostics by improving accessibility, reducing costs, and increasing testing efficiency across various genomic targets.