RT Journal Article T1 Application of MSB selective enrichment followed by amplicon MinION sequencing for multipathogen detection in smoked salmon A1 Costa Ribeiro, Ana A1 Lamas Freire, Alexandre A1 Prado Rodríguez, Marta A1 Garrido Maestu, Alejandro K1 Applied microbiology K1 Pathogens AB Millions of foodborne infections are reported yearly worldwide due to a variety of pathogens. A promising approach to tackle this issue relies on Next Generation Sequencing (NGS). In this work a novel multi-foodborne pathogen detection was developed. The method combines a selective enrichment step in a novel broth, multiplex PCR with interlaboratory-validated primers, long-read Flongle MinION sequencing, and data analysis in three cloud-based pipelines to overcome complex, command line-based bioinformatic data analyses. The method, was evaluated in salmon samples spiked with fresh, heat and cold stressed, bacterial cultures of Salmonella spp., E. coli O157:H7 and L. monocytogenes, as well as with in-house, and commercial mock communities, doped with Y. enterocolitica and thermotolerant Campylobacter spp. No major deviations from the expected results were obtained, reaching a limit of detection <10 CFU/25 g for Salmonella spp., E. coli O157 and L. monocytogenes with sensitivity, specificity and accuracy values > 90% regardless the bioinformatic pipeline selected and concordance values between 0.9 to 1.0. Taken together, the proposed method allows for simple and reliable implementation of NGS as testing tool to streamline foodborne pathogen detection, and overcoming the typical limitations associated with this technology. PB Nature SN 2396-8370 YR 2025 FD 2025-10-28 LK https://hdl.handle.net/10347/43498 UL https://hdl.handle.net/10347/43498 LA eng NO Costa-Ribeiro, A., Lamas, A., Prado, M. et al. Application of MSB selective enrichment followed by amplicon MinION sequencing for multipathogen detection in smoked salmon. npj Sci Food 9, 211 (2025). https://doi.org/10.1038/s41538-025-00452-5 NO Agenda SMARTgNOSTICS – Global Testing & Diagnostics Solutions for antimicrobial resistances, reference n.º C644915155-00000024 NO Ministry of Science, Innovation and Universities in Spain for her grant BG22/00139 - Beatriz Galindo program DS Minerva RD 27 abr 2026