Portable detection strategies for wine microorganisms using colorimetric LAMP and lateral-flow dipstick assays

Abstract

Effective detection of wine microorganisms is essential for product quality, stability and consumer safety. Culture-based techniques and molecular methods such as PCR are often expensive, time-consuming and susceptible to inhibitors present in wine matrices. This study evaluated loop-mediated isothermal amplification (LAMP) as a rapid, accurate and cost-efficient alternative for detecting microorganisms involved in wine fermentation and spoilage. Broad-range Panbacteria and Panfungal primers, together with genus- and species-specific primer sets, were designed and validated. Target organisms included Saccharomyces cerevisiae and Oenococcus oeni, which are essential for alcoholic and malolactic fermentation, and spoilage microorganisms such as Brettanomyces bruxellensis and Acetobacter aceti. Reaction conditions were optimised using additives including gold nanoparticles and mineral oil. Amplification products were visualised by turbidity, hydroxy naphthol blue and neutral red colorimetric indicators, gel electrophoresis and lateral-flow dipstick assays. All methods successfully enabled endpoint detection, providing several practical alternatives for monitoring microbial presence. The assays showed high specificity and minimal cross-reactivity with non-target organisms. LAMP therefore offers a practical and rapid tool for microbial monitoring in wineries, supporting timely intervention, wine-quality preservation and spoilage prevention.

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.2025.59.4.953
2. Ghani, A., – Ferrer, S. (2025). Loop-mediated isothermal amplification-lateral flow dipstick (LAMP-LFD) for detection of wine microorganisms. Available at SSRN 5963101

Acknowledgements

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