Abstract: Objective: The real-time isothermal Scorpio-DAMP assay for the determination of bacterial pathogens, specifically targeting commensal organisms, is important for clinical practice. Methods: The assay for Neisseria meningitidis was characterized using a set of lower respiratory tract pathogens, with sensitivity assessed at a genomic equivalent (GE) input of 100 000 for a positive reaction, and had no cross-reactivity. Results: The Scorpio-DAMP assay demonstrated a reaction time less than 30 min for positive slope recognition, with negative percent accuracy and positive percent accuracy of 91.8 and 70.6%, respectively. Discussion: Compared to the conventional PCR, which requires temperature cycling and longer processing times, the Scorpio-DAMP assay offers a more accessible solution for high-throughput and low-resource testing environments. Conclusions: The Scorpio-DAMP assay represents a promising alternative for rapid bacterial pathogen detection, particularly suitable for settings with limited resources.