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Functional Nucleic Acids for Pathogenic Bacteria Detection.

Dingran Chang, Sandy Zakaria, Sahar Esmaeili Samani

  • 1School of Environmental Science and Technology, Key Laboratory of Industrial Ecology and Environmental Engineering (Ministry of Education), Dalian University of Technology, Dalian, 116024, China.

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Summary
This summary is machine-generated.

Functional nucleic acids (FNAs) offer a rapid and reliable method for detecting infectious bacteria at the point-of-care. These novel biosensors demonstrate high sensitivity and specificity, paving the way for improved clinical diagnostics.

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Area of Science:

  • Biotechnology and Biosensing
  • Molecular Biology
  • Clinical Diagnostics

Background:

  • Pathogen detection is critical for public health, but current methods are often slow and complex.
  • Existing technologies fail to provide simple, rapid, and reliable detection at the point-of-need.
  • Functional nucleic acids (FNAs) offer unique advantages for biosensor development, including specificity, stability, and ease of modification.

Purpose of the Study:

  • To develop and demonstrate the efficacy of functional nucleic acid (FNA)-based biosensors for the rapid detection of infectious bacteria.
  • To explore the use of *in vitro* selection for isolating pathogen-specific FNAs without prior biomarker identification.
  • To integrate FNAs into point-of-care (POC) devices for direct detection in clinical samples.

Main Methods:

  • Isolation of RNA-cleaving fluorogenic DNAzymes (RFDs) and DNA aptamers using a 'many-against-many' *in vitro* selection approach against diverse bacterial targets.
  • Development of fluorescent, colorimetric, and electrochemical biosensors incorporating the isolated FNAs.
  • Integration of signal-amplification strategies (e.g., rolling circle amplification, catalytic hairpin assembly) to enhance sensitivity.
  • Testing of biosensors with clinical samples (urine, blood, stool) for direct pathogen detection.

Main Results:

  • Successfully isolated numerous pathogen-specific FNAs targeting bacteria like *Escherichia coli*, *Clostridium difficile*, *Helicobacter pylori*, and *Legionella pneumophila*.
  • Developed POC biosensors, including paper-based and hand-held devices, demonstrating high sensitivity (down to 10 cells/mL) in clinical samples.
  • Demonstrated effective signal amplification strategies to improve detection limits.
  • Identified strategies to enhance FNA stability and mitigate issues like non-specific binding and biofouling in clinical settings.

Conclusions:

  • FNA-based biosensors represent a promising platform for simple, rapid, and sensitive detection of infectious pathogens at the point-of-care.
  • The developed biosensors show significant potential for direct application in clinical diagnostics, enabling timely identification of bacterial infections.
  • Further optimization is needed to address challenges related to FNA stability and non-specific interactions in complex biological matrices for widespread clinical adoption.