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<p>Foodborne pathogens, such as <i>Escherichia coli</i> O157:H7, pose serious public health risks, highlighting the need for rapid and quantitative detection methods. Although traditional agglutination assays provide rapid results, they lack the ability to quantify analyte concentration. In contrast, gold-standard techniques such as enzyme-linked immunosorbent assays (ELISA) and polymerase chain reaction (PCR) offer high sensitivity but require complex instrumentation and lengthy procedures. In this study, we present a microfluidic chip-based quantitative agglutination assay that addresses these limitations by integrating patterned microstructures and nonspectroscopic signal processing. The chip consists of a polydimethylsiloxane (PDMS) layer bonded to a glass substrate and features repeating trapezoidal microchannels that enable passive flow control through variable channel widths. These structures also serve as standardized optical measurement zones. Detection was achieved using retroreflective Janus microparticles (RJPs) functionalized with anti-<i>E. coli</i> O157:H7 antibodies that form aggregates upon binding to the target bacteria. The size of these aggregates correlates with the cell concentration and affects their settling behavior as they flow through the channel; larger clusters settle earlier, resulting in shorter signal distances. Owing to the retroreflective properties of RJPs, these aggregates can be visualized as bright white dots using a simple and cost-effective optical setup composed of a light-emitting diode and a camera, or even by the naked eye. This approach enables sensitive and quantitative detection, improves conventional agglutination assays without the need for specialized instruments, and represents a relevant strategy for broader food safety monitoring and point-of-care testing.</p>

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Distance-Based Microfluidic Analyzer for Foodborne Pathogen Detection Using Retroreflective Janus Particle Aggregation and Non-spectroscopic Optical Readout

  • 期刊论文

  • 发表日期：

    2025年8月19日

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  • Kyung Won Lee,
  • Soo A Park,
  • Kyung Hee Song,
  • Yu Jin Sung,
  • Hyunjin Yoon,
  • Hyun C. Yoon

摘要

Foodborne pathogens, such as Escherichia coli O157:H7, pose serious public health risks, highlighting the need for rapid and quantitative detection methods. Although traditional agglutination assays provide rapid results, they lack the ability to quantify analyte concentration. In contrast, gold-standard techniques such as enzyme-linked immunosorbent assays (ELISA) and polymerase chain reaction (PCR) offer high sensitivity but require complex instrumentation and lengthy procedures. In this study, we present a microfluidic chip-based quantitative agglutination assay that addresses these limitations by integrating patterned microstructures and nonspectroscopic signal processing. The chip consists of a polydimethylsiloxane (PDMS) layer bonded to a glass substrate and features repeating trapezoidal microchannels that enable passive flow control through variable channel widths. These structures also serve as standardized optical measurement zones. Detection was achieved using retroreflective Janus microparticles (RJPs) functionalized with anti-E. coli O157:H7 antibodies that form aggregates upon binding to the target bacteria. The size of these aggregates correlates with the cell concentration and affects their settling behavior as they flow through the channel; larger clusters settle earlier, resulting in shorter signal distances. Owing to the retroreflective properties of RJPs, these aggregates can be visualized as bright white dots using a simple and cost-effective optical setup composed of a light-emitting diode and a camera, or even by the naked eye. This approach enables sensitive and quantitative detection, improves conventional agglutination assays without the need for specialized instruments, and represents a relevant strategy for broader food safety monitoring and point-of-care testing.