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Updated: Jul 5, 2025
Kinetic Screening of Nuclease Activity using Nucleic Acid Probes
Published on: November 1, 2019
Fluorescent and electrochemical detection of nuclease activity associated with Streptococcus pneumoniae using
Garazi Goikoetxea1,2,3, Khadija-Tul Kubra Akhtar1,4,5, Alona Prysiazhniuk1,4,5,6
1Nucleic Acids Technologies Laboratory (NAT-Lab), Linköping University, 58185, Sweden. frank.hernandez@liu.se.
Abstract:
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.
Insights
This study identifies a novel biomarker for detecting Streptococcus pneumoniae (S. pneumoniae) using bacterial nucleases. Developed biosensors offer rapid, specific detection of S. pneumoniae in clinical samples.
Area of Science:
- Biomarker Discovery
- Biosensor Technology
- Microbial Pathogenesis
Background:
- Streptococcus pneumoniae (S. pneumoniae) is a major cause of community-acquired pneumonia.
- Rapid and accurate detection of S. pneumoniae is crucial for timely treatment and improved patient outcomes.
- Existing diagnostic methods may lack the speed or sensitivity required for effective clinical management.
Purpose of the Study:
- To screen and discover a novel biomarker for S. pneumoniae detection.
- To develop and evaluate fluorescence and electrochemical biosensors utilizing this biomarker.
- To assess the sensitivity, specificity, and clinical applicability of the developed biosensors.
Main Methods:
- Utilized S. pneumoniae nucleases as the target biomarker.
- Identified a specific oligonucleotide substrate for the nuclease activity.
- Developed fluorescence and electrochemical biosensor platforms based on the nuclease-substrate interaction.
- Evaluated biosensor performance using bacterial cultures and clinical samples.
Main Results:
- An electrochemical biosensor achieved a high sensitivity of 10^2 CFU mL-1.
- A fluorescence biosensor demonstrated a detection limit of 10^6 CFU mL-1.
- Both biosensors exhibited high specificity, distinguishing S. pneumoniae from other pathogens.
- Successful detection of S. pneumoniae in clinical samples was achieved.
Conclusions:
- Novel S. pneumoniae nuclease-based biomarker identified.
- Electrochemical biosensor shows high sensitivity and potential for rapid diagnostics.
- Developed biosensors demonstrate clinical utility for S. pneumoniae detection.
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