Loop-mediated isothermal amplification-lateral flow dipstick (LAMP-LFD) for detection of wine microorganisms

Wine is undoubtedly one of the oldest and most celebrated beverages across cultures. It is primarily crafted from grapes through a fermentation process where yeast transforms sugars into alcohol. The presence of unwanted microorganisms can cause spoilage, so early detection of microorganisms is crucial. The previously reported conventional methods and molecular methods have so many drawbacks they are time consuming and expensive to be used in wine industries. Therefore, this study presents a novel broad-range loop-mediated isothermal amplification coupled with lateral flow dipstick (LAMP-LFD) assay for the rapid detection of wine-related microorganisms. The novelty of this work lies in the design and modification broad-range LAMP primer sets, labelled with biotin and fluorescein isothiocyanate, enabling simultaneous detection of wine spoilage microorganisms, including acetic acid bacteria and Brettanomyces fungi, as well as beneficial wine fermentation microorganisms such as lactic acid bacteria and Saccharomyces cerevisiae, with immediate visualisation of amplification products on the LFD strip. The results illustrate that the designed primers were specific for wine-related microorganisms at optimised conditions in the LAMP-LFD. The newly developed method for detecting wine-related microorganisms was very sensitive, and it can detect 10² CFU/mL, which is more sensitive than commercial polymerase chain reaction (PCR). This developed LAMP-LFD detection method is an essential advancement for the wine industry, which offers a reliable and efficient means to protect wine quality. We believe that the method is a promising proof-of-concept tool that could be further developed and validated for use in the wine industry.