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一种用于识别和定量生熟食品中肉类品种掺假的数字PCR方法。

A digital PCR method for identifying and quantifying adulteration of meat species in raw and processed food.

作者信息

Ren Junan, Deng Tingting, Huang Wensheng, Chen Ying, Ge Yiqiang

机构信息

College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, China.

Agro-product Safety Research Center, Chinese Academy of Inspection and Quarantine, Beijing, China.

出版信息

PLoS One. 2017 Mar 20;12(3):e0173567. doi: 10.1371/journal.pone.0173567. eCollection 2017.

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.

摘要

肉类掺假是一个全球性问题。本文开发了一种新的液滴数字PCR(ddPCR)方法,用于定量测定绵羊和山羊肉制品中鸡肉的存在情况。同时,引入了一个常数(倍增因子),将拷贝数的比例转化为肉的比例。所提出的ddPCR方法也被证明比实时PCR更准确(在5%至80%的范围内偏差小于9%),实时PCR已广泛用于此测定。该方法在不同热处理和超高压条件下表现出良好的重复性和稳定性。对于5%鸡肉含量,超高压或热处理的相对标准偏差(RSD)值小于5.4%。此外,我们证实肉的不同部位对ddPCR方法的定量准确性没有影响。与实时PCR相比,我们研究了ddPCR作为一种更精确、灵敏和稳定的分析策略的性能,以克服扩增效率差异方面的潜在问题,并直接获得拷贝数而无需标准曲线。本研究中开发的方法和策略不仅可用于定量绵羊肉中掺假物的存在情况并确认其不存在,还可应用于其他种类的肉类和肉制品。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/980b/5358868/54c40a019aad/pone.0173567.g001.jpg

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