Bead-based approaches to CRISPR diagnostics

Abstract

AbstractCRISPR-based diagnostics have emerged as a promising tool for fast, accurate, and portable pathogen detection. There has been rapid progress in areas such as pre-amplification processes and CRISPR-related enzymes, but the development of reporter systems and reaction platforms has lagged behind. In this paper, we develop new bead-based techniques that can help fill both gaps. First, we develop a novel bead-based split-luciferase reporter system with improved sensitivity compared to standard fluorescence-based reporter design in CRISPR diagnostics. Second, we develop a highly deployable, bead-based platform capable of detecting nine distinct viral targets in parallelized, droplet-based reactions. We demonstrate the enhanced performance of both approaches on synthetic and clinical samples. Together, these systems represent new modalities in CRISPR diagnostics with increased sensitivity, speed, multiplexing, and deployability.