Abstract
A real-time loop amplified (Rti-LAMP) DNA assay system was developed for the rapid detection of Salmonella enterica on the leaves of romaine lettuce. The assay involved seeding 25 g portions of leaves with various numbers of Salmonella CFU, binding of cells at 4°C for 18 h, followed by rinsing off the seeded cells with 50 mL of 0.025% SDS, filtration through glass wool, and pelleting the bacterial cells by centrifugation at 13,000 g. The resulting pellets were suspended in saline and processed for cell lysis and DNA purification. Purified DNA samples were dissolved in 20 μL of deionized water and 2 μL incorporated into 25 μL LAMP reactions at 65°C for 60 min. The lowest level of DNA consistently detected by the LAMP assay was that from 4 CFU/g of lettuce equivalent to the DNA from 10 CFU per LAMP reaction. The entire assay could be completed in 3 h.
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