[Rapid detection of Vibrio cholerae by loop mediated isothermal amplification (LAMP) method]

Area of Science:

• Microbiology
• Food Safety
• Molecular Diagnostics

Background:

• Vibrio cholerae is a significant foodborne pathogen causing acute intestinal illness.
• Rapid detection methods are essential for early diagnosis and effective public health interventions.
• Current detection methods may lack the speed and sensitivity required for timely outbreak management.

Purpose of the Study:

• To develop and validate a rapid, sensitive, and specific DNA-based detection method for Vibrio cholerae.
• To establish optimal conditions for a loop-mediated isothermal amplification (LAMP) assay targeting a specific Vibrio cholerae gene.
• To evaluate the performance of the developed LAMP assay in detecting Vibrio cholerae in various food samples and clinical specimens.

Main Methods:

• Design of specific primers targeting the housekeeping gene mdh of Vibrio cholerae.
• Optimization of reaction conditions for DNA loop-mediated isothermal amplification (LAMP).
• Validation of the assay's sensitivity, specificity, and performance in real-world food and clinical samples.

Main Results:

• A highly sensitive LAMP assay was successfully established with an optimal reaction at 65°C for 60 minutes.
• The detection limit was determined to be 25 CFU/mL for cultivated Vibrio cholerae and 32 CFU/g for contaminated food.
• The assay demonstrated excellent specificity, amplifying only Vibrio cholerae strains among 33 tested bacteria, and showed high agreement with ISO standards in practical sample analysis.

Conclusions:

• The developed LAMP assay is a sensitive, rapid, and simple tool for detecting Vibrio cholerae.
• This method can significantly aid in the surveillance and control of Vibrio cholerae infections.
• The assay's efficiency in detecting the pathogen in diverse food matrices and clinical samples highlights its practical utility.