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Related Experiment Video

Updated: Jun 10, 2025

Aptamer-Based Target Detection Facilitated by a 3-Stage G-Quadruplex Isothermal Exponential Amplification Reaction
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A dual-mode aptasensor based on rolling circle amplification enriched G-quadruplex for highly sensitive IFN-γ

Lianhui Zhao1, Yingai Yin1, Shuqi Xiao1

  • 1College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, 100029, China.

Analytica Chimica Acta
|October 13, 2024
PubMed
Summary

This study introduces a novel dual-mode aptasensor for highly reliable interferon-gamma (IFN-γ) detection. The innovative sensor combines colorimetric and fluorescent detection methods, enhancing accuracy and enabling cross-validation for robust results.

Keywords:
Dual-mode aptasensorG-quadruplexInterferon gammaRolling circle amplification

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Area of Science:

  • Biomedical Engineering
  • Analytical Chemistry
  • Molecular Diagnostics

Background:

  • Aptasensors offer high sensitivity and rapid detection but lack inherent reliability verification.
  • Single-mode aptasensors present challenges in validating detection accuracy in real-time.
  • Developing cross-validated detection methods is crucial for enhancing biosensor reliability.

Purpose of the Study:

  • To design and develop a colorimetric and fluorescent dual-mode aptasensor for highly reliable detection of interferon-gamma (IFN-γ).
  • To engineer G-quadruplex assembly and rolling circle amplification for enhanced sensitivity and accuracy.
  • To create a versatile platform for multimode biosensing applications.

Main Methods:

  • Modification of magnetic beads with anti-IFN-γ aptamers and complement sequences (cDNA).
  • IFN-γ triggered release of cDNAs, followed by magnetic separation.
  • Rolling circle amplification (RCA) of released cDNAs to generate G-quadruplexes.
  • G-quadruplex-hemin complex for colorimetric detection (TMB substrate) and G-quadruplex-Thioflavin T for fluorescent detection.

Main Results:

  • The dual-mode aptasensor achieved a linear detection range of 1-10000 pM (colorimetric) and 0.1-10000 pM (fluorescent).
  • Detection limits were as low as 0.406 pM (colorimetric) and 0.037 pM (fluorescent).
  • The aptasensor demonstrated satisfactory performance in detecting IFN-γ in human serum samples with good recovery rates.

Conclusions:

  • The developed dual-mode aptasensor effectively integrates aptamer binding, G-quadruplex formation, and RCA for reliable IFN-γ detection.
  • This strategy provides a robust tool for IFN-γ testing and advances the development of multimode sensing platforms.
  • The dual-mode approach enhances detection reliability through built-in cross-validation capabilities.