微滴式数字PCR对肉制品中羊肉和猪肉定量分析

为了实现肉制品中羊肉和猪肉的量化研究,本文应用微滴式数字PCR对肉制品进行定量检测。通过在看家基因上设计单拷贝特异性引物,能更好的鉴别是人为因素的掺假还是在加工过程中无意的带入。根据dd PCR的结果显示,在一定范围肉质量和DNA的浓度,DNA浓度和DNA拷贝数(C)成现一定的线性关系。通过人为掺假及市售样品检测以验证此方法。以DNA浓度作为中间换算值,得到肉质量和DNA拷贝数之间的换算关系M_羊=0.12C-10.6 M_猪=0.07C+4。通过对已知掺假比例的肉样及市售样品进行检测,能够较好的得到掺假肉的质量。目前市场上存在一定的掺假现象且该检测方法具有一定市场应用前景,本文建立的微滴式数字PCR在羊肉和猪肉及其制品中的掺假检测具有较强的应用潜力,对目前混乱的肉制品掺假的市场监管提供了科学的依据。

Quantitative Analysis of Mutton and Pork in Meat Products by Micro Digital PCR

In order to realize the quantitative research of mutton and pork in meat products, a quantitative method based on the droplet digital polymerase chain reaction (ddPCR) was used for quantitative detection of meat products in this study. It was better to identify the adulteration of human factors or inadvertent bringing into during the process by designing single primers on housekeeping genes. The results of ddPCR showed that the relationships between the DNA concentration and DNA copy number(C) were close to be linear in a certain rage of meat weight and DNA concentration. This method was validated by artificial adulteration and commercial sample detection. The DNA concentration was utilized as an intermediate value to establish the formulae about the relationship between the original meat weight and DNA copy number: Movine=0.12C-10.6, Mpork=0.07C+4. By examining the mixed meat samples of known adulteration composition and commercially available samples, a more accurate method for detecting the adulteration meat was established. It shows that there is a certain adulteration in the market, and the droplet digital polymerase chain reaction(ddPCR) established in this paper has great potential in adulteration detection of mutton and pork and their products, which provides a scientific basis for the market supervision of adulterated meat products at present.