Assessing Concordance between RNA-Seq and NanoString Technologies in Ebola-Infected Nonhuman Primates Using Machine Learning

This study evaluates the concordance between RNA sequencing (RNA-Seq) and NanoString technologies for gene expression analysis in non-human primates (NHPs) infected with Ebola virus (EBOV). We performed a detailed comparison of both platforms, demonstrating a strong correlation between them, with Spearman coefficients for 56 out of 62 samples ranging from 0.78 to 0.88, with a mean of 0.83 and a median of 0.85. Bland-Altman analysis further confirmed high consistency, with most measurements falling within 95% confidence limits. A machine learning approach, using the Supervised Magnitude-Altitude Scoring (SMAS) method trained on NanoString data, identified OAS1 as a key marker for distinguishing RT-qPCR positive from negative samples. Remarkably, when applied to RNA-Seq data, OAS1 also achieved 100% accuracy in differentiating infected from uninfected samples using logistic regression, demonstrating its robustness across platforms. Further differential expression analysis identified 12 common genes including ISG15, OAS1, IFI44, IFI27, IFIT2, IFIT3, IFI44L, MX1, MX2, OAS2, RSAD2, and OASL which demonstrated the highest levels of statistical significance and biological relevance across both platforms. Gene Ontology (GO) analysis confirmed that these genes are directly involved in key immune and viral infection pathways, reinforcing their importance in EBOV infection. In addition, RNA-Seq uniquely identified genes such as CASP5, USP18, and DDX60, which play key roles in immune regulation and antiviral defense. This finding highlights the broader detection capabilities of RNA-Seq and underscores the complementary strengths of both platforms in providing a comprehensive and accurate assessment of gene expression changes during Ebola virus infection.