Multi-platform rapid quantification of wine microorganisms: smartphone, microplate reader, real-time qLAMP and PMA-qLAMP approaches

Abstract

The rapid and accurate quantification of wine microorganisms is essential for quality control and spoilage prevention. Traditional culture-based methods, PCR and flow cytometry can be time-consuming and costly, whereas loop-mediated isothermal amplification (LAMP) offers rapid detection at low cost. This study developed and validated a multi-platform strategy combining smartphone-assisted endpoint analysis, a microplate reader, real-time quantitative LAMP (qLAMP) and propidium monoazide qLAMP (PMA-qLAMP). Specific primers targeting representative spoilage yeasts and bacteria, including Brettanomyces bruxellensis and Acetobacter aceti, were designed. Endpoint quantification of hydroxy naphthol blue-coloured amplicons was performed with a microplate reader and a user-friendly smartphone application. Real-time qLAMP was used for quantitative analysis, while PMA-qLAMP enabled viable cells to be distinguished from dead cells. Smartphone and microplate-reader assays provided rapid and cost-effective semi-quantification. qLAMP showed a strong correlation with conventional plate counts (R² > 0.95), and PMA-qLAMP reduced false-positive signals from non-viable cells by more than 90 %. The system was reproducible across the different platforms. These methods provide wineries with efficient, portable and low-cost alternatives for quantifying microorganisms during winemaking.

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. 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. DOI: 10.3389/fmicb.2018.01945

Acknowledgements

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