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Abstract
In this project, a highly precise quantitative method based on the digital polymerase chain reaction (dPCR) technique was developed to determine the weight of pork and chicken in meat products. Real-time quantitative polymerase chain reaction (qPCR) is currently used for quantitative molecular analysis of the presence of species-specific DNAs in meat products. However, it is limited in amplification efficiency and relies on standard curves based Ct values, detecting and quantifying low copy number target DNA, as in some complex mixture meat products. By using the dPCR method, we find the relationships between the raw meat weight and DNA weight and between the DNA weight and DNA copy number were both close to linear. This enabled us to establish formulae to calculate the raw meat weight based on the DNA copy number. The accuracy and applicability of this method were tested and verified using samples of pork and chicken powder mixed in known proportions. Quantitative analysis indicated that dPCR is highly precise in quantifying pork and chicken in meat products and therefore has the potential to be used in routine analysis by government regulators and quality control departments of commercial food and feed enterprises.
Highlights
In 2013, the horse meat adulteration scandal [1, 2] swept across Europe, a region which is considered to hold the highest food safety standards worldwide
BlastN searches of the entire NCBI genome database were used to validate the specificity of the primers, probes, and polymerase chain reaction (PCR) amplicons; all of the pork- and chickenspecific PCR primers, probes, and PCR amplicons bore a high level of species specificity
Through a well-designed experiment, this study has demonstrated that the digital polymerase chain reaction (dPCR) technique can be used to accurately quantify the weight of specific meats in meat products
Summary
In 2013, the horse meat adulteration scandal [1, 2] swept across Europe, a region which is considered to hold the highest food safety standards worldwide. In this scandal, meat foods advertised as containing beef were found to contain undeclared meat, such as horse meat. As much as 100% of the meat content was other undeclared meats. Due to the similar textures and processing technologies of different meat products, it is difficult for consumers to identify their authenticity, not to mention the precise proportion of meats within a given product. Reliable qualitative and quantitative detection methods need to be researched and developed to make this task easier
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