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Microbial Biosensors01:17

Microbial Biosensors

Microbial biosensors are analytical devices that utilize living microbes to detect specific substances through measurable signals. These devices consist of two main components: biosensing organisms and signal-transducing elements. Biosensing organisms, such as Escherichia coli or Saccharomyces cerevisiae, are typically housed in multiwell plates connected to transducers, enabling rapid, real-time detection of target analytes.Signal Generation MechanismWhen a target analyte—such as...

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On Column Binding a Real-Time Biosensor for β-lactam Antibiotics Quantification.

Shahla M Abdullah1,2, Shwan Rachid3

  • 1Medical Laboratory Science Department, College of Science, University of Raparin, Ranyia 46012, Sulaymaniyah, Iraq.

Molecules (Basel, Switzerland)
|March 14, 2020
PubMed
Summary

This study introduces a rapid, sensitive real-time PCR assay for detecting penicillin G antibiotic residues. The novel method accurately quantifies trace amounts in various samples, offering a practical tool for antibiotic detection.

Keywords:
Bocillin FLantibioticsbiosensormolecular quantificationpenicillinpenicillin-binding protein

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Area of Science:

  • Analytical Chemistry
  • Biotechnology
  • Pharmacology

Background:

  • Accurate detection of penicillin G residues is crucial for food safety and clinical diagnostics.
  • Existing methods for antibiotic residue detection can be time-consuming and lack sensitivity.
  • Penicillin G detection relies on its interaction with penicillin-binding proteins (PBPs).

Purpose of the Study:

  • To develop rapid, accurate, and practical tools for detecting penicillin G antibiotic residues.
  • To establish novel assays based on the binding mechanism of beta-lactam antibiotics to PBPs.
  • To compare the performance of a real-time PCR-based method with an SDS-PAGE method for penicillin G detection.

Main Methods:

  • Developed assays utilizing in vitro expressed 6X-Histidine-tagged soluble penicillin-binding protein (PBP2x*) from Streptococcus pneumoniae.
  • Employed fluorescent-labeled beta-lactam analogue Bocillin FL as a competitive substrate.
  • Utilized a real-time PCR-based fluorescence detection method and SDS-PAGE with fluorescence quantification.

Main Results:

  • Both developed methods demonstrated a broad range of linearity and high sensitivity.
  • The real-time PCR-based method detected penicillin G in the range of 0.01-0.2 nM with high accuracy.
  • The SDS-PAGE method showed sensitivity ranging from 0.015 to 2 µM.

Conclusions:

  • The on-column real-time PCR assay is a fast, highly sensitive, and accurate method for detecting penicillin G.
  • This novel assay, based on binding inhibition using a fluorescent competitor, can be adapted for screening penicillin G in biological and environmental samples.
  • The developed real-time PCR method offers a significant improvement in speed and sensitivity compared to the SDS-PAGE method.