Efficient DNA Extraction Coupled with Fluorescence Loop-Mediated Isothermal Amplification (LAMP) for On-Site Early...

Vibrio parahaemolyticus (V. parahaemolyticus), a significant pathogen in aquatic products, necessitates rapid and precise detection to prevent foodborne disease outbreaks. Early detection of bacterial pathogens is crucial for effective prevention and control. In this study, we developed a field-deployable, "sample-in, answer-out" workflow that integrates efficient DNA extraction with visual fluorescence loop-mediated isothermal amplification (LAMP) for the on-site detection of V. parahaemolyticus. This extraction protocol enables efficient one-pot DNA release within 10 min, eliminating the need for additional purification steps. In addition, coupled with an optimized fluorescence LAMP assay targeting V. parahaemolyticus, the entire process-from sample to result-can be completed within approximately 50 min. The optimized assay exhibited high sensitivity, detecting as low as 1 copy/µL of plasmid DNA and 10 CFU per reaction from pure bacterial culture and spiked seawater samples. This extraction-amplification integrated approach demonstrates robust performance and high practicality for on-site early warning and pathogen monitoring in aquaculture safety settings.