Determination of content of camel milk in adulterated milk samples by normalized real-time polymerase chain reaction system based on single-copy nuclear genes

文献类型： 外文期刊

第一作者： Wang, Zhiying

作者： Wang, Zhiying;Li, Tingting;Yu, Wenjie;Qiao, Lu;Liu, Rui;Zhao, Yan;Yang, Shuming;Chen, Ailiang;Li, Shanshan;Li, Shanshan

作者机构：

关键词： real-time PCR; quantitative determination; camel milk in adulterated milk samples; single-copy genes; normalized system

期刊名称：JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE （ 影响因子：3.638； 五年影响因子：3.802 ）

ISSN： 0022-5142

年卷期：

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收录情况： SCI

摘要： BACKGROUND Compared with the traditional qualitative polymerase chain reaction (PCR), which only identifies the category of species, the quantitative PCR method provides a value, which is very important for appropriate penalty enforcement according to the extent of adulteration. However, most of the current quantitative PCR methods are based on mitochondrial genes, expressing different copy numbers in different cells and reducing the accuracy of quantitative results. In this study, single-copy nuclear housekeeping genes, instead of multicopy mitochondrial genes, were selected as both camel species-specific and reference genes to develop a novel normalized PCR system. RESULTS This system had an excellent linear correlation (R-2 = 0.9614) between camel milk content and Ct ratio (specific/reference genes), and allowed quantitative determination of the content of camel milk in adulterated milk samples. The accuracy was effectively validated using simulated adulterated samples with recoveries ranging from 90% to 120% and coefficient of variation less than 10%, exhibiting sufficient parameters of trueness and precision. CONCLUSIONS The normalized PCR system based on single-copy nuclear genes is a simple, rapid and reliable method for the determination of the content of camel milk in adulterated milk samples, and also provides technical support for appropriate penalty enforcement. (c) 2020 Society of Chemical Industry

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