Rapid Detection of Salmonella spp from Meat: Loop Mediated Isothermal Amplification (LAMP)

Abstract

Loop-mediated isothermal amplification (LAMP) is a novel, high specific and sensitive method which amplifies nucleic acid under isothermal conditions. Salmonella is considered one of the threatening pathogens in food industries and these species are associated with distinct food poisoning called salmonellosis. Four primers (two outer and two inner primers) were designed to target six distinct regions on the target gene invA which is conserved in Salmonella species. The reaction was optimised for 60 mins at 65 °C. The sensitivity of the LAMP and PCR assay for Salmonella was 10 CFU/ml and 100 CFU/ml respectively. Artificial spiking of chicken meat shows detection of Salmonella even at dilution to extinction (<1 CFU/ml) immediately after spiking as well after 48hr enrichment. All the LAMP experiments were compared to PCR method. This study reports the development of a highly sensitive, specific and a rapid diagnostic assay for the detection of Salmonella from food. The developed method could be very useful for routine pathogens point of care (POC) diagnostics.