Goikoetxea, GaraziAkhtar, Khadija-tul KubraPrysiazhniuk, AlonaBorsa, Barış A.Aldağ, Mehmet ErsoyKavruk, MuratÖzalp, Veli CengizHernandez, Frank J.Basic SciencesNutrition and Dietetics2024-07-082024-07-0820241364-5528https://hdl.handle.net/20.500.14411/6481Open Access; Published by Analyst; DOI https://doi.org/10.1039/D3AN01532G; Garazi Goikoetxea, Nucleic Acids Technologies Laboratory (NAT-Lab), Linköping University, Sweden, Department of Cellular Biology and Histology, Faculty of Medicine and Odontology, University of Basque Country (UPV/EHU), Spain, SOMAprobes SL. Donostia, Spain; Khadija-Tul Kubra Akhtar, Frank J. Hernandez, Nucleic Acids Technologies Laboratory (NAT-Lab), Linköping University, Sweden, Wallenberg Center for Molecular Medicine, Linköping University, Sweden, Department of Physics, Chemistry and Biology, Linköping University, Sweden; Alona Prysiazhniuk, Nucleic Acids Technologies Laboratory (NAT-Lab), Linköping University, Sweden, Wallenberg Center for Molecular Medicine, Linköping University, Sweden, Department of Physics, Chemistry and Biology, Linköping University, Sweden, Taras Shevchenko National University of Kyiv, Kyiv, Ukraine; Baris A. Borsa, Nucleic Acids Technologies Laboratory (NAT-Lab), Linköping University, Sweden, Wallenberg Center for Molecular Medicine, Linköping University, Sweden, Department of Physics, Chemistry and Biology, Linköping University, Sweden; Mehmet Ersoy Aldag, Prof. Dr A. Ilhan Özdemir State Hospital, Giresun, Turkey; Murat Kavruk, Department of Medical Biology, School of Medicine, Istanbul Aydin University, Istanbul, Turkey; Veli C. Ozalp, Department of Medical Biology, Atilim University, Ankara, Turkey.Streptococcus pneumoniae (S. pneumoniae) represents a significant pathogenic threat, often responsible for community-acquired pneumonia with potentially life-threatening consequences if left untreated. This underscores the pressing clinical need for rapid and accurate detection of this harmful bacteria. In this study, we report the screening and discovery of a novel biomarker for S. pneumoniae detection. We used S. pneumoniae nucleases as biomarker and we have identified a specific oligonucleotide that works as substrate. This biomarker relies on a specific nuclease activity found on the bacterial membrane, forming the basis for the development of both fluorescence and electrochemical biosensors. We observed an exceptionally high sensitivity in the performance of the electrochemical biosensor, detecting as low as 102 CFU mL−1, whereas the fluorescence sensor demonstrated comparatively lower efficiency, with a detection limit of 106 CFU mL−1. Moreover, the specificity studies have demonstrated the biosensors’ remarkable capacity to identify S. pneumoniae from other pathogenic bacteria. Significantly, both biosensors have demonstrated the ability to identify S. pneumoniae cultured from clinical samples, providing compelling evidence of the potential clinical utility of this innovative detection system.enArticle