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Digital recombinase polymerase amplification chip based on asymmetric contact angle composite interface.

Zhongping Zhang1, Tianwei Li1, Yao Tan2

  • 1Institute of Microfluidic Chip Development in Biomedical Engineering, College of Information Science and Technology, Beijing University of Chemical Technology, Beijing, 100029, China.

Analytica Chimica Acta
|January 12, 2025
PubMed
Summary

We developed a novel asymmetric contact angle digital isothermal detection chip to improve digital recombinase polymerase amplification (dRPA) accuracy. This technology enhances reagent flow and reduces signal aggregation for more reliable nucleic acid quantification.

Keywords:
Asymmetric contact angle composite interfaceDigital RPAFluorescence aggregationMicrowell array

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Area of Science:

  • Biotechnology
  • Molecular Diagnostics
  • Microfluidics

Background:

  • Digital recombinase polymerase amplification (dRPA) is crucial for nucleic acid quantification and rare mutation detection.
  • High reagent viscosity and physical properties hinder dRPA accuracy and reproducibility.
  • Wider adoption of dRPA is limited by challenges in current amplification platforms.

Purpose of the Study:

  • To develop an optimized asymmetric contact angle digital isothermal detection (ACA-DID) chip.
  • To enable rapid and robust digital recombinase polymerase amplification.
  • To overcome limitations associated with viscous reagents in dRPA.

Main Methods:

  • Designed a pressure-driven, asymmetric contact angle composite interface for digital RPA.
  • Incorporated surfactants into PDMS to create asymmetric contact angles, improving reagent flow.
  • Optimized RPA reagent composition and employed surface modification (PDMS doped with surfactant) to enhance amplification efficiency.

Main Results:

  • The ACA-DID chip design facilitates robust digital RPA, even with viscous reagents.
  • Surfactant-modified PDMS surfaces improve reagent flow and prevent adsorption of biological macromolecules.
  • Reduced signal aggregation and improved amplification uniformity were achieved.
  • Demonstrated excellent analytical accuracy in quantifying African swine fever DNA.

Conclusions:

  • The ACA-DID chip effectively addresses key limitations in digital RPA platforms.
  • This innovation enhances the accuracy and reproducibility of nucleic acid detection.
  • The technology shows potential for broader application, especially in resource-limited settings.