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

Area of Science:

• Microbiology
• Biotechnology
• Food Science

Background:

• Wine production relies on microbial fermentation but is susceptible to spoilage by unwanted microorganisms.
• Conventional and molecular detection methods for wine microbes are often time-consuming and expensive.
• Rapid and accurate microbial detection is crucial for maintaining wine quality and preventing spoilage.

Purpose of the Study:

• To develop a novel, rapid, and sensitive assay for the simultaneous detection of key wine-related microorganisms.
• To address the limitations of existing methods in terms of speed, cost, and efficiency for the wine industry.

Main Methods:

• Design and modification of broad-range loop-mediated isothermal amplification (LAMP) primer sets.
• Labeling primers with biotin and fluorescein isothiocyanate for enhanced detection.
• Coupling LAMP with lateral flow dipstick (LFD) for immediate visualization of amplification products.
• Testing specificity and sensitivity against spoilage and beneficial wine microorganisms.

Main Results:

• The developed broad-range LAMP-LFD assay demonstrated high specificity for target wine microorganisms under optimized conditions.
• The assay achieved a sensitive detection limit of 10^2 CFU/mL, surpassing commercial polymerase chain reaction (PCR) sensitivity.
• Simultaneous detection of spoilage (acetic acid bacteria, Brettanomyces) and beneficial (lactic acid bacteria, Saccharomyces cerevisiae) microbes was achieved.

Conclusions:

• The novel LAMP-LFD assay provides a reliable, efficient, and sensitive tool for detecting wine-related microorganisms.
• This method represents a significant advancement for the wine industry, aiding in quality control and spoilage prevention.
• The developed assay shows promise as a validated tool for routine use in wine production environments.