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Implementation of Microfluidics for Antimicrobial Susceptibility Assays: Issues and Optimization Requirements
Nicole C Parsley1, Amanda L Smythers1, Leslie M Hicks1
1Department of Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, NC, United States.
Frontiers in Cellular and Infection Microbiology
|October 12, 2020
Summary
Comparing microfluidic and traditional plate assays for antimicrobial peptides (AMPs), this study found significant differences in bioactivity profiles. Method development is crucial for microfluidics to effectively detect AMPs from natural products.
Area of Science:
- Biochemistry and Molecular Biology
- Microbiology
- Bioanalytical Chemistry
Background:
- The rise of multi-drug resistant pathogens necessitates novel antimicrobial agents, with natural product peptides being a promising source.
- Traditional bioactivity assays for antimicrobial peptides (AMPs) are limited by sample quantity requirements and optimization challenges.
- Microfluidic devices present a potential advancement over conventional bulk assay systems for AMP discovery.
Purpose of the Study:
- To compare the bioactivity profiles of microfluidic and traditional 96-well plate assay formats.
- To evaluate the effectiveness of microfluidic assays for detecting AMPs from natural product libraries.
- To identify challenges and requirements for implementing microfluidic devices in AMP screening.
Main Methods:
- A microfluidics-based bioassay was developed and compared against a standard 96-well plate format.
- A peptide library from *Viola inconspicua* was screened against *E. coli* ATCC 25922 in both assay types.
- Bioactivity in microfluidics was assessed via brightfield microscopy; plate assays used optical density and fluorescence measurements.
Main Results:
- Both assay formats detected bioactivity in specific *V. inconspicua* peptide fractions, though with some variations in optical density and fluorescence readings.
- Significant differences were observed in the bioactivity profiles between the microfluidic and plate-based assays.
- Potential reasons for discrepancies include differences in assay materials and growth/assay conditions.
Conclusions:
- While microfluidics offer potential for AMP detection, their direct application requires substantial method development and optimization.
- The materials and conditions inherent to each assay format significantly influence measured antimicrobial peptide bioactivity.
- Further research is needed to realize the full potential of microfluidic devices for sensitive and rapid AMP screening from natural products.
