Detection of the Bovine-derived Materials and Their Contents in Meat Products by Real-time Fluorescence Quantitative PCR

TAN Bo; YANG Chun-ping; LIU Xiao-ling

doi:10.13651/j.cnki.fjnykj.2023.10.002

Abstract：In order to establish a rapid,efficient,accurate and low-cost method for the identification and quantitative detection of the bovine-derived materials in meat products,the primers and probes were synthesized with the single-copy gene of cell nucleus,bosPDE as the target gene and LcoR as the internal reference gene,to optimize the reaction conditions and evaluate the specificity and sensitivity of the method.By using the relative quantitative method of real-time fluorescence PCR,the regression curves were respectively established with the linear model of △Ct value and 2△△Ct value,and the accuracy of the method was evaluated by simulating artificially the mixed beef samples.The results showed that:the bosPDE primer of the target gene only amplified the DNA of beef,which was specific,and the sensitivity of amplification to beef DNA could reach 0.01 ng·μL-1.The regression curves with △Ct value and 2△△Ct value as the quantitative indexes were y=-3.096 5x+8.479 7(R2=0.998)and y=0.712 2x+3.050 4(R2=0.988 7),respectively.The recovery rates of the artificial simulated mixed beef samples with 35％-90％beef con-tent detected by the two methods were 94.30％-102.52％and 98.55％-106.30％,respectively.The deviation of recovery rate was reduced when the number-average treatment was carried out on the two quantitative methods.The real-timefluorescence quantitative PCR method established in this study could accurately characterize the bovine-de-rived materials in meat products,and could be used for the relative quantitative analysis of beef in the beef adulterated products.It had important reference value for identifying the adulteration of meat products and quantifying the con-centrations of components in beef products.