EBOV-specific gene signature identified in nonhuman primates and human cohorts with Ebola infection

IntroductionEbola virus (EBOV) infection triggers intense host transcriptional responses that overlap extensively with those induced by other viral and bacterial pathogens. This overlap complicates the identification of EBOV-specific gene expression signatures and limits diagnostic specificity. Defining transcriptional markers that distinguish EBOV from other infections is essential for improving molecular diagnostics and advancing understanding of EBOV-specific host responses.MethodsWe developed a multi-step filtering framework using blood-derived RNA-Seq data from nonhuman primates and human cohorts organized into independent training and test sets. In the training cohort, differential expression analysis was performed using an edgeR-based GLMQL-MAS approach to identify EBOV-associated genes. Candidates were filtered against non-EBOV comparator datasets, including mpox virus, influenza, bacterial pneumonia, acute HIV-1 infection, and multiple SARS-CoV-2 variants, to remove broadly shared host-response genes. Genes included in the NanoString nCounter® Host Response Panel were additionally excluded. The resulting EBOV-specific signature was evaluated in independent EBOV and non-EBOV test cohorts using principal component analysis and logistic regression. Functional enrichment was assessed using KEGG pathways.ResultsInitial analysis identified numerous interferon-stimulated genes that were similarly upregulated across infections. After cross-infection filtering and NanoString exclusion, 281 EBOV-specific genes were identified. Optimization within the training cohort yielded a top-50 gene set that clearly separated EBOV from Non-EBOV samples. In the independent test cohort, classification performance improved substantially, with the F1 score increasing from 37.5% when all genes were used to 95.0% after applying the top-50 gene set. Enrichment analysis of the top-50 EBOV-specific genes revealed significant association with vascular, coagulation, secretory, and metabolic pathways. ADAMTS1 showed consistent upregulation in EBOV while remaining downregulated or inactive in comparator infections.DiscussionStructured cross-pathogen filtering enables identification of EBOV-specific transcriptional features beyond shared antiviral responses. The validated gene signature generalizes across independent cohorts and highlights biologically distinct pathways, which supports its potential utility for host-based diagnostic development.