Digital PCR Detection Method For Five Animal-derived Components In Meat Products

According to media reports and relevant functional departments,it is reported that the phenomenon of “doping and making false” in meat products on the market is very serious and has become a major issue facing the food industry in China.The research on the identification methods of meat products is very necessary,and therefore relevant.The inspection of meat products by the department must be strengthened.It is imperative to establish relevant testing standards to provide evidence for the identification and tracking of adulteration in meat products.In this study,specific primers and probes for micro-drop digital PCR were designed based on the specific gene sequences of chicken,cattle,sheep,pig,and duck sources to establish meat products containing different chickens,cattle,and sheep.Quantitative micro-droplet PCR assay for the detection of porcine,duck,and duck-derived components.The establishment of this method provides a reliable technical basis for the relevant law enforcement agencies.In the study on the animal-derived components of chickens,cattle,sheep,pigs and ducks in meat products,the specificity of the five animal-derived components was tested.PCR amplification results showed that the five animal-derived The specificity of primers and probes was good,and there were no homologous amplification of common livestock and poultry species including horse meat,rabbit meat,donkey meat and wheat flour.Different quality meat samples were processed to optimize DNA extraction methods.Take different gradients and a meat-like quality of DNA extracted meat samples,to establish a meat-like quality-meat-like DNA standard curve concentration.Different dilutions of the extracted DNA were performed,and the copy number of the droplets was measured to establish a standard curve of DNA concentration-number of digital PCR copies.Using DNA concentration as an intermediate variable,a standard curve between meat quality-number of digital PCR copies was established.For the established standard curve,the total mass of 100 mg was used to verify the minimum detection limits of 1%,5%,10%,and 50% of the known meat samples,and thestability of the minimum detection limit was verified.he results showed that the meat sample content could be detected at 5% or more.Under the premise of keeping the total content unchanged,anti-interference experiments were performed on meat samples of 5%,10%,50%,70% and 90% known to be added with different kinds of meat samples.The results showed that other types of meat on the surface had little effect on known meat samples.n the study of the detection of animal-derived components in meat products,the established methods were used,and samples known to be commercially available were tested.series of meat products such as chicken fillets,ducks,beef rolls,roast leg of lamb and barbecued meat were tested and the results showed that they can effectively detect different animal-derived components in different foods.The establishment of micro-drop digital PCR quantitative detection technology enables more rapid and effective detection of animal-derived components with specificity and rapid and accurate methods.