Detection of Bovine Components in Meat Products Based on Real-time Fluorescence Quantitative PCR and Digital PCR
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Abstract:
In order to detect bovine-derived components in meat products quickly and effectively, real-time fluorescence quantitative PCR and digital PCR were used to detect bovine-derived components in meat products. Firstly, the specific primers and probes were designed by multiple sequence alignment according to the specific region of beef single replication genome, then the standard genomic DNA was extracted by phenol-chloroform method. Real-time fluorescence quantitative PCR and digital PCR reaction conditions were established. The purity and concentration of DNA template were determined and the extracted DNA solution was free of impurities. On this basis, the fluorescence quantitative analysis of bovine-derived components was constructed. The specificity, anti-interference and bovine-derived components in meat products were detected by standard curve of PCR. The results showed that both primers and probes have fluorescence signals for beef. The two methods have bovine origin specificity, the detection values are basically consistent with the actual sample values. The two methods have good anti-interference ability. When there are other meat disturbances, the bovine origin can still be accurately monitored. The maximum errors of the two methods were 3.8% and 4%, respectively. The bovine origin components in different meat products can be quantitatively detected. The bovine origin components in 10 beef balls of samples can not be detected by all methods, which verifies the market. Fake meat does exist. It showed that the proposed method can accurately and rapidly detect bovine-derived ingredients in meat products, and is suitable for the detection of meat products in the market. It is of great significance to protect the rights and health of consumers.
