Detection of Salmonella in Food Using ttrA Gene Optimized Loop-Mediated Isothermal Amplification Technology
Objective:The objective of this research is to optimize the loop-mediated isothermal amplification(LAMP)detection technology for faster and more accurate detection of Salmonella which could provide the technical support for the prevention and control in food.Methods:A redesigned LAMP detection primer was developed based on the ttrA gene found in the ttrRSBCA transposon of Salmonella,the primer's were validated through experiments testing its range,specificity,and sensitivity,and it was compared to invA1,invA2,invA3,and bcfD primers,as well as two commercially available D and S brand detection kits.Results:The redesigned primers ttrA,ttrB-2,and ttrA120-1 demonstrated strong stability in detecting Salmonella.Additionally,primers ttrA120-1,bcfD,and a commercially available S brand detection kit exhibited 100％exclusivity for 14 non-Salmonella species in this detection.And the exdusivity of primer ttrA and commercially available D test kits was 64.7％and 85.7％.The primers of invA1,invA3 exhibited 0％exclusivity and poor specificity.The primer ttrA120-1 could effectively detect 23 Salmonella species across 26 different serotypes with an inclusiveness rate of 88.5％.Similarly,the primer ttrA can detect 22 species with an inclusiveness rate of 84.6％.The primer bcfD had a detection inclusiveness rate of 73.1％for 19 species.Commercially available D and S test kits could detect 19 and 20 Salmonella species with inclusiveness rates of 73.1％and 76.9％.The ttrA120-1 primer was able to detect DNA concentrations as low as 3.2×10-5 ng/μL in sensitivity experiments.Conclusion:The primer ttrA120-1 has higher sensitivity,specificity,and detection ability compared to commercially available kits and published primer sequences,making it a more accurate and efficient tool for the rapid detection of Salmonella in food.
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