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Developing a dual-RCA microfluidic platform for sensitive E. coli O157:H7 whole-cell detections.

Yuqian Jiang1, Zhenyu Qiu2, Tao Le3

  • 1Department of Chemical and Biological Engineering, University of Ottawa, 161 Louis Pasteur, Ottawa, ON, K1N 6N5, Canada; Measurement Science and Standards, National Research Council Canada, 100 Sussex Drive, Ottawa, ON, K1A 0R6, Canada; Key Laboratory of Food Nutrition & Safety, Ministry of Education, Tianjin University of Science & Technology, Tianjin, 300457, China.

Analytica Chimica Acta
|August 18, 2020
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Summary

A new dual-rolling circle amplification (RCA) method enhances aptamer microfluidic platforms for sensitive whole cell detection. This approach significantly improves both target capture and signal amplification for food safety applications.

Keywords:
AptamersAtomic force microscopyBiosensorsFoodborne pathogen detectionMicrofluidic channelsRolling circle amplification

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Area of Science:

  • Microfluidics
  • Biotechnology
  • Food Safety

Background:

  • Aptamer-based microfluidic platforms are rapidly advancing.
  • Improving detection sensitivity is crucial for these platforms.
  • Whole cell detection in microfluidic devices requires enhanced strategies.

Purpose of the Study:

  • To develop a dual-rolling circle amplification (RCA) approach for sensitive whole cell detection using microfluidic devices.
  • To enhance both target cell capture and signal amplification for improved detection limits.

Main Methods:

  • A dual-RCA strategy combining capturing RCA (cRCA) and signaling RCA (sRCA).
  • cRCA modifies microfluidic channels with poly-aptamers for enhanced target cell capture.
  • sRCA amplifies detection signals compared to single fluorescence probes.

Main Results:

  • Poly-aptamer channels captured 3-fold more target cells than mono-aptamer channels.
  • sRCA enhanced detection signals up to 50-fold compared to single probes.
  • The dual-RCA system achieved a combined signal enhancement of approximately 250-fold.

Conclusions:

  • The dual-RCA microfluidic device offers a simple and promising approach for sensitive whole-cell detection.
  • This method is effective in various food matrices like orange juice and milk.
  • A limit of detection of 80 cells/mL for E. coli O157:H7 was achieved, supporting food safety inspections.