A New Method For Detection Of Adulterated Duck Meat Based On PCR Amplification Platform

Beef is rich in protein,which is a kind of meat favored by Chinese residents.However,the price of beef is expensive.Many illegal traders mix low-priced meat such as duck into beef for profits,which not only violates the interests of consumers but also disturbs the market order.Therefore,it is particularly important to establish an effective detection method for beef adulteration.In this paper,two detection methods for identifying duck components in beef were established based on PCR combined with lateral chromatographic immunoassay and fluorescence polarization technology.The main contents are as follows:(1)Visual detection of adulterated duck in beef by PCR combined with lateral chromatographic immunoassayThis method uses two groups of functionally-modified duck and beef primers to obtain functionally-modified amplification products.Beef amplification products can be combined with SA coupled on the surface of gold nanoparticles and SA fixed on the NC membrane to form a sandwich structure,which is fixed on the C line.Duck amplification products can be combined with SA coupled on the surface of gold nanoparticles and FITC antibody fixed on the NC membrane to form another sandwich structure,so as to be fixed on the T line.According to the color intensity of T line,we can test the adulterated duck in the beef semi-quantitatively.The detection limit of this method was 0.05 %(w/w).(2)Quantitative detection of adulterated duck meat in beef by PCR combined withThe method utilizes the change of FP value of FITC modified duck primers after PCR amplification to detect the adulterated duck meat in beef.In the experiment,FITCmodified primers were used as fluorescent signal molecules.In the absence of duck DNA,the molecular weight of the amplified duck primers were unchanged after amplification,and the FP value was relatively small.In the presence of duck DNA,FITC-modified primers were amplified into double-stranded DNA products with large molecular weight,and the FP value was significantly increased.Meanwhile,there was a linear relationship between the change in FP value and the adulteration ratio of duck meat in beef samples.Therefore,the proportion of the adulterated component can be directly calculated according to the value of FP,realizing the quantitative detection of duck adulteration in beef samples.The detection limit of this method was 0.1 %(w/w).