Towards deployable CRISPR-based nucleic acid detection.

Abstract

Deployable diagnostics are necessary for the control and treatment of infectious diseases, with significant unmet needs revealed during the COVID-19 pandemic. Nucleic acid diagnostics remain among the most sensitive and specific forms of detection, yet their reliance on laboratory equipment and trained personnel limits their deployment in resource limited settings. CRISPR-based diagnostics are uniquely positioned to enable rapid, affordable, and highly accurate nucleic acid testing at both the point-of-care and the point-of-need. In this review, we discuss advances toward deployable CRISPR-based diagnostics. We begin by examining innovations in sample processing methods, emphasizing strategies that reduce equipment requirements and enhance compatibility across diverse sample types and pathogens. We then explore developments in one-pot isothermal and amplification-free approaches, comparing the benefits and tradeoffs associated with each, as well as multiplexing strategies for simultaneous detection of multiple pathogens. Finally, we consider additional factors that impact assay deployability, including reagent lyophilization to minimize cold chain dependence and readout technologies that enable detection in resource-limited settings. We conclude by outlining remaining challenges and opportunities for future progress.