Study on Detection Method for Pig,Cow and Sheep Derived Components by Multiple Fluorescence PCR in Meat Skewer Products
[Objective]To establish a multiple fluorescence PCR detection method for common cattle,sheep and pig derived components in an-imal meat skewer products,and to achieve rapid identification of animal derived components in meat skewer products.[Method]Compared the extraction effect of DNA from meat mince tissue using magnetic bead method and column membrane method,optimized the real-time fluores-cence PCR multiple detection method,and compared the amplification effect of cattle,sheep,and pig derived components at two annealing tem-peratures of 58 and 60℃.[Result]The two DNA extraction methods used in the experiment had similar effects,and the A260/A280 values of the DNA solution could meet the requirements of PCR detection.58℃ was the optimal annealing temperature,at which point the cattle,sheep and pig derived DNA could be effectively amplified,and the lowest detected DNA concentration was 10-3 ng/μL.The primers and probes used in the method had good specificity and had not produced non-specific amplification.[Conclusion]The multiplex PCR detection method established by the application scope experiment for cattle,sheep and pig origin can provide a method reference for rapid qualitative analysis of animal origin components in meat products.
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