ddPCR-based profiling of plasma EV-miRNA identifies a noninvasive biomarker panel for tuberculosis

Background Tuberculosis (TB) is a top infectious disease killer; underdiagnosis fuels its spread. Thus, finding perspective non-invasive biomarkers for early tuberculosis (TB) is very essential. Extracellular vesicles (EVs) microRNAs (miRNAs) are promising non-invasive biomarkers for tuberculosis (TB) detection. Methods To investigate the feasibility of early diagnosis of TB, we examined plasma-derived EV-miRNAs obtained from patients with active TB (n = 25), asymptomatic tuberculosis (aTB) (n = 25), and latent tuberculosis infection (LTBI) (n = 25). Initially, EVs were isolated from plasma samples using ultracentrifugation and were characterized via nanoparticle tracking analysis and immunoblotting for surface markers. Additionally, we characterized miRNA profiles using RNA sequencing to identify TB-specific EV-miRNAs. EV-miRNAs were further validated by droplet digital PCR (ddPCR). Results Seven differentially expressed miRNAs among the clinical subgroups (fold change > 2, P Conclusion This multigroup cohort study established a novel EV-miRNAs signature and demonstrated its clinical utility for TB staging and early detection.