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Updated: Sep 11, 2025

Aptamer-Based Target Detection Facilitated by a 3-Stage G-Quadruplex Isothermal Exponential Amplification Reaction

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Ultrasensitive and Multiplexed Target Detection Strategy Based on Photocleavable Mass Tags and Mass Signal

Seokhwan Ji¹, Jin-Gyu Na¹, Woon-Seok Yeo¹

¹Department of Bioscience and Biotechnology, Bio/Molecular Informatics Center, Konkuk University, Seoul 05029, Republic of Korea.

Nanomaterials (Basel, Switzerland)

|August 13, 2025

Summary

This study introduces a mass spectrometry platform for detecting multiple viral co-infections simultaneously. The novel system enhances sensitivity and accuracy for diagnosing infections like hepatitis B virus (HBV) and HIV.

Keywords:

hybridization chain reaction mass spectrometry mass tags multiplexed detection photocleavable signal amplification

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Area of Science:

Analytical Chemistry
Biotechnology
Molecular Diagnostics

Background:

Co-infections present significant clinical and diagnostic challenges.
Accurate detection of multiple viral pathogens requires sensitive, multiplexed analytical platforms.
Current methods may lack the sensitivity or multiplexing capability for complex biological samples.

Purpose of the Study:

To develop a mass spectrometry (MS)-based platform for simultaneous detection of multiple viral targets.
To enhance signal amplification using target-triggered hybridization chain reaction (HCR).
To enable quantitative analysis using in situ photocleavable mass tags (PMTs).

Main Methods:

Engineered hairpin DNAs with PMTs and loop structures on magnetic particles (Loop@MPs) for each target.
Utilized native polyacrylamide gel electrophoresis (PAGE) and laser desorption/ionization MS (LDI-MS) for validation.
Employed silica@gold core-shell hybrid (SiAu) nanoparticles as an internal standard for quantitative reliability.

Main Results:

Achieved high sensitivity with a detection limit of 415.12 amol for hepatitis B virus (HBV).
Demonstrated a broad dynamic range from 1 fmol to 100 pmol.
Successfully detected and quantified mixtures of HBV, human immunodeficiency virus (HIV), and hepatitis C virus (HCV) with high accuracy and precision in fetal bovine serum.

Conclusions:

The developed MS-based platform offers robust and reliable ultrasensitive multiplexed detection.
The system shows strong potential for clinical diagnostics and biomedical research applications.
This approach addresses the critical need for simultaneous detection of viral co-infections.