Establishment Of Fluorescent Quantitative PCR Detection Method For Duck-Derived Components In Meat And Meat Products

In recent years,there have been many cases of food adulteration and forgery driven by economic interests in the domestic and foreign markets.The identification of food ingredients is of great significance for protecting consumer rights and maintaining market order.The adulteration of meat products requires identification of the DNA in the sample.Nucleic acid-based detection methods can identify genus and subgenus levels and have become the mainstream technology for animal-derived component detection and species identification.In this study,duck species-specific primers and probes were designed and screened,and real-time fluorescent PCR was applied to explore the identification and quantitative detection methods of duck-derived components in meat products.The main research contents and conclusions are as follows:?1?Bioinformatics was used to break the genomic DNA sequence of duck species into 100 bp fragments,and a single copy sequence unique to duck species was screened out with the method of BLAST sequence comparison,screening de-duplication,length filtering and other steps,combined with various animal genomic DNA information and genome annotation information.Primers and Taqman probes were designed in the specific sequence interval.The specificity of the primers was verified by routine PCR,and two pairs of high specific primers,Anapl₁ and Anapl₂,were obtained.?2?The genomic DNA of 16 animals and 3 plants was used as the templates and the real-time fluorescent PCR technique was applied for specific verification,reaction condition optimization,sensitivity investigation,detection limit analysis and repetitive test.The qualitative detection method of duck-derived components was established.The detection target fragment is only 69 bp,which is suitable for the detection of various meat products,especially processed foods.The results showed that the duck-derived detection system established in this study has high specificity and stability,and the sensitivity can reach 10^-3 ng/?L duck-derived DNA.The detection limit is 0.1%?w/w?duck meat content.?3?Introduce the internal reference gene and construct the fluorescence quantitative standard curve,and establish a quantitative detection method for duck-derived components.The constructed standard curve R² is above 0.99 and has good amplification efficiency.The average recovery rate of the quantitateve detection of duck-derived components in the simulated samples was up to 96.67%,and the detection results were highly accurate.This method was used to qualitatively and quantitatively detect duck-derived components in 15 commercial samples.The results showed that the method established in this study can effectively identify duck-derived components in commercial meat products,and the test results can be used as a reference for adulteration identification,which has a good practicability.