Rapid and user-friendly detection and quantification of wine microorganisms using LAMP

Abstract

The rapid detection and quantification of microorganisms is essential for monitoring fermentation, preventing microbial spoilage, and preserving wine quality. This study focuses on the development of loop-mediated isothermal amplification (LAMP)-based assays to identify and quantify yeasts and bacteria of oenological relevance, including Saccharomyces cerevisiae, Oenococcus oeni, Brettanomyces bruxellensis, and Acetobacter aceti. Broad-range, genus-specific, and species-specific primers were designed, and reaction conditions were optimised using additives such as gold nanoparticles and mineral oil. Amplification products were detected using turbidity measurements, colorimetric indicators such as hydroxy naphthol blue and neutral red, gel electrophoresis, and lateral flow dipstick assays. Quantification was performed in various ways including smartphone- and microplate reader-based colorimetric analysis, as well as real-time quantitative LAMP and PMA-qLAMP, which enables the differentiation of viable and non-viable cells. The assays demonstrated high specificity, minimal cross-reactivity, and good reproducibility. qLAMP results were strongly correlated with conventional plate counts (R² > 0.95). Furthermore, PMA-qLAMP reduced false-positive signals from dead cells by over 90 %. These approaches provide wineries with portable, rapid, and cost-effective tools for microbiological monitoring and timely decision-making throughout the winemaking process.

References

1. Ghani, A., & Ferrer, S. (2025). Optimised loop-mediated isothermal amplification (LAMP) for reliable detection and quantification of lactic acid bacteria in red and white wines and musts. OENO One, 59(4). https://doi.org/10.20870/oeno-one.2025.59.4.9531
2. Ghani, A., & Ferrer, S. (2025). Loop-mediated isothermal amplification-lateral flow dipstick (LAMP-LFD) for detection of wine microorganisms. Available at SSRN 5963101.
3. Soares-Santos et al. (2018). Direct and rapid detection and quantification of Oenococcus oeni cells in wine by cells-LAMP and cells-qLAMP. Frontiers in Microbiology, 9, 1945. https://doi.org/10.3389/fmicb.2018.01945
4. Ferrer, S. Rapid and User-Friendly Detection and Quantification of Wine Microorganisms Using LAMP. Recording of the presentation delivered at Enoforum Spain 2026, Zaragoza, 20–21 May 2026. Available soon on Infowine Premium: https://vinidea.it/es/brand/infowine/

Acknowledgements

This research was supported by the Higher Education Commission, Government of Pakistan (HEC).