数字聚合酶链反应
重复性
食品科学
生肉
熟肉
相对标准差
实时聚合酶链反应
数学
生物
动物种类
聚合酶链反应
生物系统
计算生物学
检出限
统计
遗传学
基因
动物
作者
Junan Ren,Tingting Deng,Wensheng Huang,Ying Chen,Yiqiang Ge
出处
期刊:PLOS ONE
[Public Library of Science]
日期:2017-03-20
卷期号:12 (3): e0173567-e0173567
被引量:94
标识
DOI:10.1371/journal.pone.0173567
摘要
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.
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