Establishment And Application Of A Method For Beef Authentication Based On DNA Barcoding

Beef adulteration has always been a concern for various reasons,such as public health,religious factors,food safety,wholesomeness,and unhealthy competition in meat market.Consumers should be protected from these illegal practices of meat adulterations by quick,precise,and species authentication of meat.In recent years,DNA barcoding have been proposed as a powerful tool for identifying the species origin in meat and meat products,due to its high specificity and sensitivity,as well as simplicity and hot stability.The present study established a method for beef authentification,using the technology of DNA Barcoding and PCR-RFLP.Muscle tissue of nine species,including cattle,goat,pig,chicken,silkie,duck,fish,rat and shrimp,was collected for DNA extraction.A single universal primer pair(LCO1490/HCO2198)was adopted to amplify COI gene of the mitochondrial DNA.A single restriction enzyme(Hpa II)was selected for digestion of PCR amplification products.According to the result of agarose gel electrophoresis(3%),each species has its own species-specific restriction fragments,revealing a simultaneous identification of cattle and other animal species.Verification test was made in the laboratory to identify the effectiveness of the method for beef authentification.The method was applied for adulteration detection of commercial meat products.The method was tested for laboratory verification.Mimic counterfeiting of beef and other meat was made for testing.Species of meat mixed with beef can be detected through the method.The method can detect the doping amount as low as 1 mg and DNA samples as low as 0.1 ng/ul.Also,The method can be used for heat treated meat samples.The method was applied for authenticity test of commercial meat products.120 beef products,including beef,steak,beef liver,beef jerky,beef rolls,beef balls,beef sausage and 42 other meat products were randomly purchased from the market.9 of 120(7.5%)beef products were identified to be adulterated.In addition,this method can be used for authenticity identifying of meat products,including mutton,pork,chicken,fish,shrimp mince and duck.