Exploiting virulence determinants for precision management of melioidosis: A multi-omics roadmap from pathogenesis to...

Melioidosis, caused by Burkholderia pseudomallei, remains challenging to diagnose promptly, partly due to limited use of virulence factors as diagnostic targets. Most existing biomarkers fail to capture the dynamic expression of bacterial virulence factors during active infection. In vivo‑upregulated virulence determinants, such as Hcp1, POMCR/PPEP, and GroEL, are intrinsically superior diagnostic targets. Hcp1 is a paradigm: it is the most advanced virulence factor translated into a point‑of‑care test, with prospective validation in endemic settings. Mechanistic understanding of virulence regulation transforms empirical antigen hunting into rational target selection. We propose a virulence‑driven pipeline that converted Hcp1 into a point‑of‑care immunochromatography test (sensitivity 88.3%, specificity 86.1-100%) with prospective validation. Finally, multi‑omics data integration with explainable machine learning can prioritize virulence factor combinations for multiplex diagnostics. This framework addresses the need for rapid melioidosis diagnostics and provides a replicable template for other infectious diseases caused by intracellular pathogens.