Abstract
Meat adulteration is a worldwide concern. In this paper, a new droplet digital PCR (ddPCR) method was developed for the quantitative determination of the presence of chicken in sheep and goat meat products. Meanwhile, a constant (multiplication factor) was introduced to transform the ratio of copy numbers to the proportion of meats. The presented ddPCR method was also proved to be more accurate (showing bias of less than 9% in the range from 5% to 80%) than real-time PCR, which has been widely used in this determination. The method exhibited good repeatability and stability in different thermal treatments and at ultra-high pressure. The relative standard deviation (RSD) values of 5% chicken content was less than 5.4% for ultra-high pressure or heat treatment. Moreover, we confirmed that different parts of meat had no effect on quantification accuracy of the ddPCR method. In contrast to real-time PCR, we examined the performance of ddPCR as a more precise, sensitive and stable analytical strategy to overcome potential problems of discrepancies in amplification efficiency discrepancy and to obtain the copy numbers directly without standard curves. The method and strategy developed in this study can be applied to quantify the presence and to confirm the absence of adulterants not only to sheep but also to other kinds of meat and meat products.
Highlights
Adulteration of meat products occurs frequently worldwide [1,2,3]
The average value of the k value for five different meat fractions was 0.8, and the relative standard deviation (RSD) was 9.9%, demonstrating that the k values were stable against any mass ratio of chicken and sheep
These results demonstrated that the sensitivity of the developed droplet digital PCR (ddPCR) method was better than that
Summary
Adulteration of meat products occurs frequently worldwide [1,2,3]. The condition occurs such as deliberately adding cheaper derivatives to bulk out minced meat products, unwitting adulteration by poor manufacturing methods, or mislabeling (deliberate or unwitting) of one meat as another. To protect the interests of consumers, many countries and regions have issued regulations regarding the labeling of meat products [6,7,8], and several methods for the identification of animal species based on DNA [9,10,11,12] or protein (peptides) [13,14,15,16] have been developed to coordinate with the implementation. Determining of the ratio of adulteration remains a critical issue because it is difficult to discriminate whether the adulterants were included deliberately or inadvertently
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