Rapid, Amplification-free, RNA-based Uropathogen Detection With LbuCas13a.

Background: Expeditious identification of bacterial infection remains an important challenge in an emergency department. Bacterial cultures remain the gold standard, though they take 24-72 hours to result. Polymerase chain reaction-based diagnostics are emerging but take several hours to get a result. Here, we report a rapid bacterial RNA detection platform based on Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) technology with urine-to-result time within 1 hour. Methods: All CRISPR experiments were conducted as an open format plate reader assay with fluorescent readouts. In derivation studies, Results: The lower limit of detection of our CRISPR assay was ∼10 Conclusions: We demonstrated the feasibility to adopt an amplification-free CRISPR assay for the purpose of rapid uropathogen detection. To our knowledge, this is the first demonstration of an RNA-based tool for detecting uropathogens. Our assay may be used postanalytically to conventional urinalysis for improved specificity to diagnose UTIs. Future research may focus on improving the sensitivity and discriminating uropathogen versus bacterial contaminant, which is beyond the scope of the current study.