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Facile prepared microfluidic chip for multiplexed digital RT-qPCR test.

Wenqi Hu1,2, Yidan Zhu2, Qu Tang3

  • 1Department of Laboratory Medicine, Affiliated Hospital of Nantong University, Nantong, Jiangsu, P. R. China.

Biotechnology Journal
|September 13, 2023
PubMed
Summary

This study introduces a simple centrifugal microfluidic chip for digital polymerase chain reaction (PCR), enabling high-throughput nucleic acid quantification. The innovative design simplifies fabrication and achieves accurate results across various concentrations.

Keywords:
SARS-CoV-2influenza virusmicrofluidicmultiplexed digital RT-qPCRthe chip-based digital PCR

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Area of Science:

  • Biotechnology
  • Microfluidics
  • Molecular Diagnostics

Background:

  • Chip-based digital PCR is crucial for nucleic acid amplification and quantification in molecular diagnostics.
  • Previous digital PCR chip designs face challenges in fabrication, surface properties, and throughput, limiting widespread adoption.

Purpose of the Study:

  • To develop a facile digital microfluidic chip for digital PCR analysis driven by centrifugal force.
  • To overcome limitations of existing digital PCR platforms, focusing on simplified fabrication and high-throughput capabilities.

Main Methods:

  • A novel digital microfluidic chip utilizing centrifugal force for reagent manipulation and droplet generation.
  • Immiscible phase (silicone oil) encapsulation of PCR reagents within 9600 microchambers for digital partitioning.
  • Centrifugal priming approach for efficient sample loading and high-throughput processing.

Main Results:

  • Achieved efficient digitization regardless of chip surface properties (hydrophilic/hydrophobic).
  • Demonstrated high-throughput analysis with 9600 microchambers.
  • Accurate nucleic acid quantification with strong correlation (R² = 0.99) from 1 to 1000 copies/μL.
  • Showcased potential for multiplexed analysis and multi-target detection.

Conclusions:

  • The developed centrifugal digital microfluidic chip offers a simplified and high-throughput solution for digital PCR.
  • This technology has significant potential for advancing molecular diagnostics and multi-target detection applications.