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Updated: Aug 2, 2025
Visualization of Bacterial Resistance using Fluorescent Antibiotic Probes
Published on: March 2, 2020
Synthesis of vancomycin fluorescent probes that retain antimicrobial activity, identify Gram-positive bacteria, and
Bing Zhang1, Wanida Phetsang1, M Rhia L Stone1
1Centre for Superbug Solutions, Institute for Molecular Bioscience, The University of Queensland, Brisbane, QLD, 4072, Australia.
Abstract:
Antimicrobial resistance is an urgent threat to human health, and new antibacterial drugs are desperately needed, as are research tools to aid in their discovery and development. Vancomycin is a glycopeptide antibiotic that is widely used for the treatment of Gram-positive infections, such as life-threatening systemic diseases caused by methicillin-resistant Staphylococcus aureus (MRSA). Here we demonstrate that modification of vancomycin by introduction of an azide substituent provides a versatile intermediate that can undergo copper-catalysed azide-alkyne cycloaddition (CuAAC) reaction with various alkynes to readily prepare vancomycin fluorescent probes. We describe the facile synthesis of three probes that retain similar antibacterial profiles to the parent vancomycin antibiotic. We demonstrate the versatility of these probes for the detection and visualisation of Gram-positive bacteria by a range of methods, including plate reader quantification, flow cytometry analysis, high-resolution microscopy imaging, and single cell microfluidics analysis. In parallel, we demonstrate their utility in measuring outer-membrane permeabilisation of Gram-negative bacteria. The probes are useful tools that may facilitate detection of infections and development of new antibiotics.
Insights
New fluorescent vancomycin probes were synthesized to detect Gram-positive bacteria and measure outer-membrane permeabilization. These tools aid in the discovery of novel antibacterial drugs to combat antimicrobial resistance.
Area of Science:
- Medicinal Chemistry
- Microbiology
- Chemical Biology
Background:
- Antimicrobial resistance poses a significant global health threat, necessitating the development of new antibacterial agents and research tools.
- Vancomycin, a crucial glycopeptide antibiotic, treats Gram-positive infections, including those caused by methicillin-resistant Staphylococcus aureus (MRSA).
Purpose of the Study:
- To develop novel fluorescent vancomycin probes for bacterial detection and research.
- To create versatile tools for advancing antibacterial drug discovery and development.
Main Methods:
- Modification of vancomycin with an azide substituent to create a versatile intermediate.
- Copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction with alkynes to synthesize fluorescent probes.
- Evaluation of probe antibacterial activity, detection capabilities (plate reader, flow cytometry, microscopy, microfluidics), and outer-membrane permeabilization assays.
Main Results:
- Facile synthesis of three vancomycin fluorescent probes with retained antibacterial profiles.
- Demonstrated versatility of probes for detecting and visualizing Gram-positive bacteria using multiple advanced techniques.
- Successful application of probes in measuring outer-membrane permeabilization in Gram-negative bacteria.
Conclusions:
- The synthesized vancomycin fluorescent probes are effective tools for bacterial detection and research.
- These probes can aid in the early detection of infections and accelerate the development of new antibiotics to combat antimicrobial resistance.
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