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Modular Microfluidic Sensor Integrating Nucleic Acid Extraction, CRISPR/Cas13a, and Electrochemiluminescence for

Xinyuan Mao1,2, Yao Lu1,2, Zixi Gao1,2

  • 1MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, College of Biophotonics, School of Optoelectronic Science and Engineering, South China Normal University, Guangzhou 510631, China.

Analytical Chemistry
|February 28, 2025
PubMed
Summary

This study presents a novel microfluidic sensor for rapid RNA detection using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas13a and electrochemiluminescence (ECL). The integrated system achieves sensitive pathogen detection in under 30 minutes.

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Area of Science:

  • Biomedical Engineering
  • Molecular Diagnostics
  • Sensor Technology

Background:

  • Rapid and accurate pathogen screening is vital for effective disease detection and management.
  • Existing methods for RNA detection can be time-consuming and complex, hindering point-of-care applications.

Purpose of the Study:

  • To develop a modular microfluidic sensor for integrated nucleic acid extraction, clustered regularly interspaced short palindromic repeats (CRISPR)/Cas13a reaction, and electrochemiluminescence (ECL) detection.
  • To achieve rapid, sensitive, and multiplexed RNA detection for pathogen identification.

Main Methods:

  • Construction of a modular microfluidic sensor with distinct nucleic acid processing and detection modules.
  • Utilized immiscible filtration for rapid nucleic acid extraction and magnetic force for RNA distribution.
  • Integrated multichannel CRISPR/Cas13a reactions with a novel multichannel closed bipolar electrode-based ECL (MCBPE-ECL) system for signal amplification and detection.

Main Results:

  • The sensor successfully integrated RNA extraction and detection within 30 minutes.
  • Achieved a low detection limit of 0.372 fM for *Escherichia coli* 16S rRNA and 63.8 cfu/mL in human blood.
  • Demonstrated multiplexed detection by simultaneously monitoring the growth curves of *E. coli* and *Staphylococcus aureus*.

Conclusions:

  • The developed microfluidic sensor offers a rapid, sensitive, and multiplexed platform for RNA detection.
  • This technology has significant potential for improving pathogen screening and disease diagnostics.