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Lately, much effort has been made to find mRNA biomarkers for tuberculosis (TB) disease/infection with microarray-based approaches. In a pilot investigation, through RNA sequencing technology, we observed a prominent modulation of  DOCK9, EPHA4 , and  NPC2  mRNA abundance in the blood of TB patients. To corroborate these findings, independent validations were performed in cohorts from different areas. Gene expression levels in blood were evaluated by quantitative real-time PCR (Brazil,  n  = 129) or reanalysis of public microarray data (UK:  n  = 96; South Africa:  n  = 51; Germany:  n  = 26; and UK/France:  n  = 63). In the Brazilian cohort, significant modulation of all target-genes was observed comparing TB vs. healthy recent close TB contacts (rCt). With a 92% specificity,  NPC2  mRNA high expression ( NPC2  high ) showed the highest sensitivity (85%, 95% CI 65%–96%; area under the ROC curve [AUROC] = 0.88), followed by  EPHA4  (53%, 95% CI 33%–73%, AUROC = 0.73) and  DOCK9  (19%, 95% CI 7%–40%; AUROC = 0.66). All the other reanalyzed cohorts corroborated the potential of  NPC2  high  as a biomarker for TB (sensitivity: 82–100%; specificity: 94–97%). An  NPC2  high  profile was also observed in 60% (29/48) of the tuberculin skin test positive rCt, and additional follow-up evaluation revealed changes in the expression levels of  NPC2  during the different stages of  Mycobacterium tuberculosis  infection, suggesting that further studies are needed to evaluate modulation of this gene during latent TB and/or progression to active disease. Considering its high specificity, our data indicate, for the first time, that  NPC2  high  might serve as an accurate single-gene biomarker for TB.

Transcriptomic Biomarkers for Tuberculosis: Evaluation of DOCK9. EPHA4, and NPC2 mRNA Expression in Peripheral Blood