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Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
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Biosensor for Detection of Antibiotic Resistant Staphylococcus Bacteria
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Aptamer based high throughput colorimetric biosensor for detection of staphylococcus aureus.

Tianxiao Yu1, Hong Xu1, Yan Zhao1

  • 1Department of clinical laboratory, the fourth hospital of Shijiazhuang (Shijiazhuang gynaecology and obstetrics hospital), Shijiazhuang, China.

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|June 10, 2020
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Summary

This study presents a rapid, high-throughput colorimetric biosensor for detecting Staphylococcus aureus (SA). The novel aptamer-based assay achieves sensitive and specific SA quantification, offering a powerful tool for pathogen screening.

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Area of Science:

  • Biotechnology
  • Biosensor Development
  • Analytical Chemistry

Background:

  • Accurate and rapid detection of pathogenic bacteria like Staphylococcus aureus (SA) is crucial for public health.
  • Existing methods for SA detection can be time-consuming or lack high-throughput capabilities.
  • Development of sensitive and specific biosensors is essential for clinical diagnostics, food safety, and environmental monitoring.

Purpose of the Study:

  • To develop a high-throughput colorimetric biosensor for the sensitive and specific detection of Staphylococcus aureus (SA).
  • To utilize aptamer recognition and DNA-catalyzed signal amplification for colorimetric bacterial quantification.
  • To establish a rapid assay suitable for simultaneous analysis of multiple samples.

Main Methods:

  • Immobilization of SA-specific aptamers on a 96-well plate via streptavidin-biotin binding.
  • Displacement of aptamer by SA, followed by hybridization with a three-way junction (TWJ) probe.
  • Photo-catalyzed colorimetric detection using the dsDNA-SYBR Green I (SG I) complex to oxidize TMB.

Main Results:

  • The biosensor achieved a limit of detection (LOD) of 81 CFU mL⁻¹ for SA in PBS buffer within 5.5 hours.
  • High specificity for SA was demonstrated, with minimal response to Escherichia coli and Pseudomonas aeruginosa.
  • The system supports simultaneous detection of 96 samples, enabling high-throughput analysis.

Conclusions:

  • The developed aptamer-based colorimetric biosensor offers a sensitive, specific, and rapid method for SA detection.
  • The high-throughput capability makes it suitable for screening large numbers of samples in various applications.
  • This assay represents a promising tool for pathogenic microorganism screening in clinical diagnostics, food safety, and environmental monitoring.