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Fluorescence Method-Based Nonlinear Hybridization Chain Reaction for Highly Sensitive and Specific Detection of HER2.

Feifei Liu¹, Jing Li¹, Wenna Jiang¹

¹Department of Laboratory, Tianjin Medical University Cancer Institute & Hospital, National Clinical Research Center for Cancer, Tianjin's Clinical Research Center for Cancer, Tianjin's Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin, China, tijmu.edu.cn.

International Journal of Analytical Chemistry

|July 9, 2026

Summary

A new, highly sensitive fluorescence assay using nonlinear hybridization chain reaction (NHCR) offers accurate, noninvasive detection of serum human epidermal growth factor receptor 2 (HER2) for gastric cancer monitoring.

Keywords:

Human epidermal growth factor receptor 2 fluorescence sensor gastric cancer nonlinear hybrid chain reaction

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Area of Science:

Biomedical Engineering
Analytical Chemistry
Molecular Diagnostics

Background:

Human epidermal growth factor receptor 2 (HER2) is a key biomarker for gastric cancer targeted therapy.
Current tissue-based tests (IHC, FISH) are gold standards, but serum HER2 offers a less invasive monitoring option.
Existing serum HER2 assays lack the sensitivity required for effective disease monitoring.

Purpose of the Study:

To develop a highly sensitive fluorescence assay for detecting serum HER2.
To utilize nonlinear hybridization chain reaction (NHCR) coupled with a HER2-specific aptamer for enhanced signal amplification.
To provide a novel tool for accurate and noninvasive serum HER2 detection.

Main Methods:

Development of a fluorescence assay employing nonlinear hybridization chain reaction (NHCR) and a HER2-specific aptamer (HB5).
NHCR facilitates branching-mediated exponential amplification for nonlinear signal growth, surpassing linear HCR.
Validation of the assay's performance using standard solutions and clinical serum samples.

Main Results:

The assay demonstrated a wide linear range (0.5-90 ng·mL⁻¹) with a low detection limit of 38 pg·mL⁻¹.
High specificity, reproducibility, and stability were observed for HER2 detection.
Results from clinical samples showed strong correlation with the chemiluminescence immunoassay (CLIA), with recoveries from 95.4% to 109.6%.

Conclusions:

The NHCR-based fluorescent assay provides a sensitive and accurate method for serum HER2 detection.
This approach offers a valuable, noninvasive tool for monitoring gastric cancer patients.
The assay is characterized by simple operation and cost-effectiveness.