Establishment of Insulated Isothermal PCR Detection Method for Salmonella in Food
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Abstract:
In order to make up for the shortcomings of traditional culture method such as long time-consuming and cumbersome on-site detection steps, an insulated isothermal PCR (iiPCR) rapid detection method for Salmonella in food was established. Specific primers and probes were designed according to the invA gene of Salmonella; bacterial DNA was quickly extracted by the water bath method; the amount of primers, probes and templates was also optimized. Then, a method for rapid detection of Salmonella based on iiPCR was established. The specificity, sensitivity and stability of the established method were evaluated. At the same time, the established method was compared with the traditional PCR method and the traditional culture method for detecting Salmonella contamination in actual food samples. The established iiPCR detection method had good specificity, high sensitivity, and no cross-reaction with other bacteria. The lowest detection limit could reach 75 CFU/mL. The established iiPCR method could detect Salmonella contaminated in actual food samples within 6 hours, while the same detection effect was achieved by traditional PCR methods at least 12 hours. The accuracy of iiPCR method was verified by the traditional culture method. The established iiPCR method for detecting Salmonella contaminated in food is suitable for field detection, more quickly and simpler.
