Integrating epidemiological evidence and immunodiagnostic development for early diagnosis of Bothrops spp....

Snakebite envenoming remains a major public health issue in Brazil, with Bothrops genus responsible for most cases. To support public health planning, we analyzed epidemiological data from Paraná State and reported, for the first time, a species-level identification of snakes responsible for accidents in this region. The results revealed that Bothrops jararaca accounted for over 85% of cases, followed by Bothrops jararacussu, reinforcing their epidemiological and clinical relevance. Based on this evidence, two monoclonal antibodies were produced by hybridoma technology for application in a diagnostic tool for early identification of Bothrops spp. envenomation. The antibodies were sequenced and immunochemically characterized, and both specifically recognized bothropic metalloproteinases. When applied in a competitive ELISA, the assay detected venom concentrations as low as 60 ng/mL in sera spiked with B. jararaca and Bothrops alternatus and showed no reactivity with other medically relevant genera. Although detection of B. jararacussu, Bothrops moojeni and Bothrops neuwiedi required higher concentrations, these venoms remained detectable, indicating potential for broader application. Further optimization could enhance its sensitivity, enabling more effective detection across a wider spectrum of species. Together, these findings provide novel epidemiological and immunodiagnostic insights, that can guide the development of improved diagnostic platforms for Bothrops spp. envenoming.