A novel platform to accelerate antimicrobial susceptibility testing inNeisseria gonorrhoeaeusing RNA signatures

The rise of antimicrobial-resistant pathogens can be attributed to the lack of a rapid pathogen identification (ID) or antimicrobial susceptibility testing (AST), resulting in delayed therapeutic decisions at the point of care. Gonorrhea is usually empirically treated with no AST results available before treatment, thus contributing to the rapid rise in drug resistance. Herein we present a rapid AST platform using RNA signatures for Neisseria gonorrhoeae (NG). RNA-seq followed by bioinformatic tools were applied to explore potential markers in the transcriptome profile of NG upon minutes of azithromycin exposure. Validation of candidate markers using PCR showed that two markers ( arsR (NGO1562) and rpsO ) can deliver accurate AST results across 14 tested isolates. Further validation of our cutoff in comparison to MIC across 64 more isolates confirmed the reliability of our platform. Our RNA markers combined with emerging molecular point-of-care systems has the potential to greatly accelerate both ID and AST to inform treatment.