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FEN1-Aided RPA (FARPA) Coupled with Autosampling Microfluidic Chip Enables Highly Multiplexed On-Site Pathogen

Yi Ma1, Yuanmeng Wang1, Chen Chen1

  • 1Department of Clinical Pharmacy, Jinling Hospital, Affiliated Hospital of Medical School, State Key Laboratory of Analytical Chemistry for Life Science, Nanjing University, Nanjing 210002, China.

Analytical Chemistry
|March 6, 2025
PubMed
Summary
This summary is machine-generated.

A new FARPA-chip platform enables rapid, low-cost, multiplexed detection of pathogens. This easy-to-use system achieves sensitive, on-site screening of infectious diseases within 50 minutes.

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Area of Science:

  • Biotechnology
  • Molecular Diagnostics
  • Microfluidics

Background:

  • Accurate, rapid, and cost-effective infectious disease diagnosis is vital for on-site pathogen screening.
  • Current point-of-care testing (POCT) methods often struggle with multiplexing and minimal instrumentation requirements.

Purpose of the Study:

  • To develop a versatile and user-friendly platform for multiplexed pathogen detection.
  • To address the limitations of existing methods for on-site infectious disease screening.

Main Methods:

  • Combination of multiplex recombinase polymerase amplification (RPA) with an autosampling microfluidic chip (FARPA-chip).
  • Utilized universal RPA primers for unbiased amplification of multiple targets from double-stranded cDNA (ds-cDNA).
  • Integrated rapid extraction reagent for reduced turnaround time.

Main Results:

  • Sensitive detection of as low as 10 copies of target RNA per 12-plex assay for vector-borne viruses within 45 minutes.
  • No cross-talk observed between different targets.
  • Successful 9-plex detection of 6 respiratory viruses in clinical nasopharyngeal swab samples, with results consistent with RT-qPCR.
  • Turnaround time reduced to under 50 minutes with integrated extraction.

Conclusions:

  • The FARPA-chip platform offers a cost-effective solution for on-site pathogen screening with high multiplexing capability.
  • The system is theoretically capable of detecting up to 24 pathogens, meeting diverse clinical needs.
  • This technology holds promise for healthcare applications in resource-limited settings.