A multiplex LAMP-melting curve method for simultaneous detection of Salmonella spp. and Staphylococcus aureus in food was developed. The primers were designed according to the sequences of invA gene of Salmonella spp. and nuc gene of Staphylococcus aureus, and the reaction system was optimized. The amplification products were analyzed by melting curves, electrophoresis, precipitation observation and fluorescence observation as well. No false positive and false negative results emerged during tests of 9 target and 23 non-target bacteria strains, and the pathogens could be determined by melting temperature on melting curves. DNA sequencing of LAMP products showed the sequences as deduced, confirming the correct LAMP amplification. The detection limits for each pathogen were both 10 fg/μL. The tests result of 232 samples showed that 2 pieces of raw pork were contaminated by Staphylococcus aureus. This method showed high specificity, and could be used to detect foodborne pathogens rapidly.

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