Quantitative detection methods of bovine-derived ingredients in meat product by chip digital PCR

Abstract

ObjectiveIn order to accurately quantify the proportion of bovine-derived ingredients in meat products， the Mitochondrial Cytb gene of bovine for quantitatively detection of bovine ingredients by the digital PCR （Dpcr） technology was developed in this paper.MethodsThis duplex dignal PCR was developed and optimized by designing bovine-derived specific， duck-derived specific， porcine-derived specific and animal-derived universal primers and probes according to standards.ResultsGood linear relationships between the raw beef meat weight and DNA content and between the DNA content and DNA copy number. By using the dPCR method， we found good linear relationships between the raw meat weight and DNA content and between the bovine -derived DNA content and DNA copy number. Using the DNA content as an intermediate value， we established the following formulae for calculating the weight of the original raw beef meat from the specific DNA copy number： Mbeef=0.020 9C+0.676. The mixed samples of known bovine quality were tested， and the results showed that the method can accurately quantify bovine-derive dingredients， which has litter interference by foreign species. Using this method to test 10 samples of the bovine-derived products on the market， by using the ratio of universal and specific amplified copy number， it could determine whether other animal-derived components were present in the beef meat product. it was found that the content of some samples was not up to standard.ConclusionThis method can realize the quantitative detection of bovine origin components， which can be used as a basis to distinguish intentional addition from unintentional pollution and provide a strong technical guarantee for law enforcement and supervision departments.

Keywords

• bovine-derived ingredients
• chip digital pcr
• meat products
• copy number
• quantitative detection