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Related Concept Videos

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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Ultrasensitive Detection of Multiple Foodborne Pathogens Using CRISPR-Cas12a on a Finger-Actuated Microfluidic Device

Dong Hyun Han1, Sang Yong Lee2, Yurim Kim1

  • 1Department of Bio and Brain Engineering, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea.

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|December 19, 2025
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A new microfluidic device uses recombinase polymerase amplification (RPA) and CRISPR-Cas12a for rapid detection of foodborne pathogens. This reusable system offers sensitive, user-friendly point-of-care testing for enhanced food safety.

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Area of Science:

  • Biotechnology
  • Microfluidics
  • Molecular Diagnostics

Background:

  • Foodborne pathogens present significant global health and economic risks.
  • Existing detection methods can be slow, costly, or prone to cross-contamination.
  • There is a need for rapid, sensitive, and user-friendly diagnostic tools for food safety.

Purpose of the Study:

  • To develop and validate a novel microfluidic platform for detecting foodborne pathogens.
  • To integrate recombinase polymerase amplification (RPA) with CRISPR-Cas12a technology for enhanced sensitivity and specificity.
  • To design a reusable and reconfigurable finger-actuated microfluidic device with a modular pressurizing pump (MoPP) for improved usability and multiplexing capabilities.

Main Methods:

  • Development of a finger-actuated microfluidic device incorporating a modular pressurizing pump (MoPP).
  • Integration of RPA and CRISPR-Cas12a systems with target-specific CRISPR RNAs (crRNAs).
  • Optimization of RPA primer and crRNA sequences for detecting *Escherichia coli* O157:H7, *Salmonella* spp., and *Listeria monocytogenes*.

Main Results:

  • The MoPP-integrated microfluidic device demonstrated reusability and reduced cross-contamination.
  • The RPA-CRISPR-Cas12a method achieved low limits of detection (LOD) for three key foodborne pathogens in spiked milk samples.
  • LODs were 1.62 CFU/mL for *E. coli* O157:H7, 1.84 CFU/mL for *Salmonella* spp., and 1.01 CFU/mL for *L. monocytogenes*.

Conclusions:

  • The developed microfluidic RPA-CRISPR-Cas12a platform is a sensitive and user-friendly tool for foodborne pathogen detection.
  • The MoPP enhances device reusability and enables flexible, multiplexed assay workflows.
  • This technology holds promise for point-of-care testing throughout the food supply chain, improving food safety monitoring.