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Aptamer-Functionalized Gold Nanoparticle Assay for Rapid Visual Detection of Norovirus in Stool Samples
Maytawan Thanunchai1,2, Sirikwan Sangboonruang1,2,3, Natthawat Semakul4
1Division of Clinical Microbiology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, Thailand.
Biosensors
|June 25, 2025
Summary
A new visual assay uses aptamer-functionalized gold nanoparticles (AuNPs) for rapid Norovirus (NoV) detection. This simple method offers a promising tool for on-site diagnosis, especially in resource-limited settings.
Area of Science:
- Nanotechnology
- Molecular Diagnostics
- Virology
Background:
- Norovirus (NoV) is a major cause of acute gastroenteritis globally, necessitating improved diagnostic tools.
- Current Norovirus detection methods are often time-consuming, require specialized equipment, and demand technical expertise.
- There is a need for rapid, visual, and accessible diagnostic assays for Norovirus outbreaks.
Purpose of the Study:
- To develop a novel visual detection method for Norovirus capsid protein using aptamer-functionalized gold nanoparticles (AuNPs).
- To evaluate the performance of this colorimetric assay in buffer and spiked stool samples.
- To assess the potential for on-site Norovirus detection, particularly in resource-limited environments.
Main Methods:
- Functionalization of gold nanoparticles (AuNPs) with SMV25 aptamers specific to Norovirus capsid protein.
- Development of a MgCl2-induced colorimetric assay based on AuNP aggregation and dispersion.
- Quantitative analysis using absorbance ratios (A630/A520) and evaluation with spiked and real stool samples.
- Scanning transmission electron microscopy (STEM) to confirm nanoparticle behavior.
Main Results:
- The assay showed a linear relationship between absorbance ratio and Norovirus capsid protein concentration in buffer (R² = 0.9026, LOD = 9.65 ng/µL) and spiked stool (R² = 0.9170, LOD = 37.11 ng/µL).
- Performance on real stool samples indicated 77% sensitivity and 65% specificity, with highest sensitivity for Norovirus GII.2 (100%) and GII.4 (78%).
- STEM confirmed the aggregation and dispersion mechanisms of the aptamer-functionalized AuNPs.
Conclusions:
- The developed aptamer-AuNP colorimetric assay offers a simple, rapid, and visual method for Norovirus detection.
- The assay shows promise for on-site Norovirus diagnostics, especially in resource-limited settings.
- Further optimization is required to enhance performance for direct testing of complex clinical specimens.
