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Integration of FISH and Microfluidics.

Célia F Rodrigues1, Nuno F Azevedo1, João M Miranda2

  • 1LEPABE - Laboratory for Process Engineering, Environment, Biotechnology and Energy, Department of Chemical Engineering, Faculty of Engineering, University of Porto, Porto, Portugal.

Methods in Molecular Biology (Clifton, N.J.)
|February 12, 2021
PubMed
Summary

This study presents a microfluidic platform integrated with peptide nucleic acid fluorescence in situ hybridization (PNA-FISH) for rapid microbial identification, bypassing lengthy enrichment steps for faster results.

Keywords:
Fluid mechanicsMicrofluidicsMicroorganism identificationModelingPNA-FISHPathogen detection

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Area of Science:

  • Microbiology
  • Biotechnology
  • Analytical Chemistry

Background:

  • Traditional microbial identification methods often require time-consuming enrichment steps to reach detection limits.
  • Faster molecular techniques are crucial for timely microbial analysis in food and clinical settings.

Purpose of the Study:

  • To develop an integrated methodology for rapid microbial identification.
  • To combine microfluidics with PNA-FISH for enhanced detection efficiency.

Main Methods:

  • Design and fabrication of microfluidic devices using computational fluid dynamics and soft lithography.
  • Adaptation and optimization of a peptide nucleic acid fluorescence in situ hybridization (PNA-FISH) protocol for microfluidic applications.
  • Focus on Saccharomyces cerevisiae identification, with adaptability for other microorganisms.

Main Results:

  • Successful integration of a microfluidic platform with PNA-FISH.
  • Demonstrated ability to concentrate cell suspensions and accelerate identification.
  • Microfluidic designs and PNA-FISH protocol are adaptable for various microorganisms.

Conclusions:

  • The integrated microfluidic-PNA-FISH approach offers a faster alternative to conventional microbial identification methods.
  • This methodology significantly reduces the need for extensive pre-enrichment steps.
  • The system's adaptability makes it a versatile tool for microbial detection in diverse sample types.