基于qPCR技术定量检测掺假骆驼奶的方法研究
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福建省产品质量检验研究院,福建 福州 350015

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张建美(1990—),女,硕士,主管药师,主要从事食品微生物学及分子生物学检测研究。E-mail:924526521@qq.com

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福建省产品质量检验研究院院级一般项目(KY202222A)。


Quantitative Detection Method for Adulteration in Camel Milk Based on qPCR
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Fujian Inspection and Research Institute for Product Quality, Fuzhou 350015, China

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    摘要:

    【目的】为进一步完善骆驼奶行业标准,促进骆驼奶产业的可持续发展,作者基于实时荧光定量PCR(qPCR)技术建立骆驼奶掺假定量检测方法。【方法】采用全自动核酸提取法提取模板DNA,以骆驼线粒体细胞色素b基因(cytb)为特异性基因,以哺乳动物18S rRNA基因为内参基因,设计特异性引物和内参引物。基于Ct比值(骆驼特异性引物Ct值/内参引物Ct值)与骆驼奶体积分数的线性关系,建立骆驼奶的掺假定量标准曲线。利用建立的标准曲线对含体积分数35%、55%、75%骆驼奶的模拟掺假骆驼奶样品进行定量分析,计算回收率、标准差及变异系数(CV)等参数。【结果】标准曲线为y=-0.325x+1.221 5,相关系数(R2)为0.972 3,模拟掺假骆驼奶样品的平均回收率为100.71%,平均CV为5.29%。【结论】作者建立的基于实时荧光定量PCR技术的检测方法可实现掺假骆驼奶定量分析,从而判定其是否掺假及掺假比例,为骆驼奶行业的质量控制提供了技术支持。

    Abstract:

    [Objective] To improve the standards and promote the sustainable development of the camel milk industry, the author developed a quantitative detection method for camel milk components in adulterated dairy products based on real-time fluorescent quantitative PCR (qPCR). [Method] Template DNA was extracted through an automated nucleic acid extraction method. The mitochondrial cytochrome b gene (cytb) of camel was selected as the specific target gene, while the mammalian 18S rRNA gene served as the reference gene. Specific primers and reference primers were designed accordingly. According to the linear relationship between the Ct ratio (Ct value of camel-specific primer/Ct value of reference primer) and the volume fraction of camel milk, a standard curve for quantitative analysis of camel milk adulteration was established. The established standard curve was used for quantitative analysis of simulated adulterated samples containing camel milk at volume fractions of 35%, 55%, and 75%, and parameters such as recovery rate, standard deviation, and coefficient of variation (CV) were calculated. [Result] The standard curve was y=-0.325x+1.221 5, and the correlation coefficient (R2) was 0.972 3. The average recovery rate of the simulated adulterated samples was 100.71%, and the average CV was 5.29%. [Conclusion] Using real-time fluorescent quantitative PCR, the author developed a detection method capable of quantifying adulteration in camel milk. This method not only identifies adulteration but also determines its exact proportion, offering valuable technical support for maintaining quality standards in the camel milk industry.

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张建美.基于qPCR技术定量检测掺假骆驼奶的方法研究[J].食品与生物技术学报,2026,45(3):111-118.

ZHANG Jianmei. Quantitative Detection Method for Adulteration in Camel Milk Based on qPCR[J]. Journal of Food Science and Biotechnology,2026,45(3):111-118.

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  • 收稿日期:2025-04-02
  • 最后修改日期:2025-05-09
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  • 在线发布日期: 2026-07-09
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