Integrating Loop-mediated Isothermal Amplification with Lateral Flow Assay to Achieve a Highly Sensitive Method for Detecting Genome in Raw Pork

., a zoonotic foodborne pathogen prevalent in Southeast Asia, poses a substantial threat to human and animal health because of its ability to cause severe and life-threatening illnesses. To address this challenge, a rapid and highly sensitive detection platform for in raw pork was developed by integrating loop-mediated isothermal amplification (LAMP) and a lateral flow assay (LFA), LAMP-LFA. LAMP reactions targeting the glutamate dehydrogenase () gene were optimized for specific detection of within 45 min at an isothermal temperature of 65 °C. The assay exhibited marked sensitivity, with a detection limit of 100 fg for genomic DNA extracted from cultures. Notably, this method showed no cross-reactivity with other bacterial contaminants commonly found in raw pork. The resulting LAMP amplicons were effectively detected using LFA, with a test limit of 10 CFU per 25 g of raw pork. LAMP-LFA proved to be highly specific and reliable, with no false-positives detected in spiked pork samples or pork samples containing other bacterial contaminants. Due to its high sensitivity, specificity, and rapid turnaround time, the proposed technique has immense potential as a field-deployable screening test for detection in raw pork, contributing to enhanced food safety and public health protection.