Quantitative Detection Of Duck And Pork In Beef By Real - Time Fluorescent

Food safety problems have become increasingly prominent at present, and meat adulteration has become very serious; therefore, solving meat security issues have become a common challenge of our society. In order to achieve the quantitative detection of meat mixed with other meat, the primers and probes were designed according to the sequence of the single copy in genome of beef, duck and pork in the study, and were detected to verify the specificity and sensitivity. Then to determine the amount of each material and the reaction temperature of Real-time PCR system, and analysis the differences of raw and cooked beef, duck and pork; and the standard curves were established to achieve the quantitative detection of beef, duck and pork. On this basis, using the Real-time PCR systems, the standard curves and the relationships between meat mass and concentration of DNA to detect beef mixed duck, beef mixed pork and beef mixed duck and pork quantitatively.The results showed that the Real-time PCR methods for beef, duck and pork quantitative detection were rapid and accurate; the primers and probes were highly specific for DNA of beef, duck and pork, and had not cross-react with other species; and the sensitivity of the primers and probes were very high, the limit of detection for beef and pork were 0.2 ng, and was 0.02 ng for duck; there were no significant differences raw and cooked beef and pork in the detection, however, detection of raw duck was significantly different from cooked duck. Therefore, beef and pork detection established one standard curve respectively, and two standard curves were established for the detection of raw and cooked duck, the standard curves had accurate linearity, and the R2 were higher than 0.99, which were accuracy for quantitative detection. The absolute error of the detection for duck mixed in beef and pork mixed in beef were less than 5%, and for duck and pork mixed in beef at the same time was less than 10%, which test results were accurate and achieved quantitative detection of meat in the level of weight.