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中国精品科技期刊2020
朱天园,焦新雅,程书梅. 基于便携式荧光定量PCR仪定量检测驴肉中掺假鸭肉[J]. 华体会体育,2023,44(9):340−345. doi: 10.13386/j.issn1002-0306.2022080139.
引用本文: 朱天园,焦新雅,程书梅. 基于便携式荧光定量PCR仪定量检测驴肉中掺假鸭肉[J]. 华体会体育,2023,44(9):340−345. doi: 10.13386/j.issn1002-0306.2022080139.
ZHU Tianyuan, JIAO Xinya, CHENG Shumei. Detecting Donkey Meat Adulterated with Duck Meat by Portable Fluorescence Quantitative PCR[J]. Science and Technology of Food Industry, 2023, 44(9): 340−345. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022080139.
Citation: ZHU Tianyuan, JIAO Xinya, CHENG Shumei. Detecting Donkey Meat Adulterated with Duck Meat by Portable Fluorescence Quantitative PCR[J]. Science and Technology of Food Industry, 2023, 44(9): 340−345. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022080139.

基于便携式荧光定量PCR仪定量检测驴肉中掺假鸭肉

Detecting Donkey Meat Adulterated with Duck Meat by Portable Fluorescence Quantitative PCR

  • 摘要: 为建立快速方便的驴肉制品分子鉴定方法,本文以驴肉和常见的掺假肉类(鸭肉)为研究对象,筛选特异性引物和TaqMan探针,利用便携式Mini8 Plus实时荧光定量PCR仪进行灵敏度和特异性实验,通过绘制扩增标准曲线及确定驴肉和鸭肉的质量与DNA比值常数,对不同掺入比例(加入定量的鸭肉制成含量分别为20%、40%、60%、80%)的模拟样品和实际驴肉样品进行检测。结果显示,该方法对驴、鸭肉均具有良好的特异性,可以与马、猪、山羊、梅花鹿、牛、鸡、狗肉明显区分;对驴源性DNA成分的检出限为0.01 ng/μL,鸭源性DNA成分的检出限为0.1 ng/μL,对驴肉与鸭肉混合物中鸭肉成分的灵敏度为0.1%(w/w);所建立的标准曲线线性关系良好,驴肉DNA扩增标准曲线:y=−3.584x+27.003,R2=0.9982;鸭肉DNA扩增标准曲线:y=−3.538x+30.907,R2=0.9991;采用已建立的方法对35份驴肉样本进行市场试点调查,发现6份(17.1%)驴肉样本中含有鸭肉成分。以上研究结果说明,该实时荧光定量PCR方法可用于驴肉产品中其他掺假肉类(鸭)的快速、准确检测,为驴肉及其制品的市场监管和相关执法提供有力的技术保障。

     

    Abstract: To develop a fast and convenient method for the molecular identification of donkey meat products, this study selected donkey meat and duck meat (a common meat used for adulteration) as the research objects, screened specific primers and TaqMan probes, and conducted sensitivity and specificity experiments with a portable Mini8 Plus real-time fluorescence quantitative PCR instrument. By drawing the amplification standard curve and determining the mass/DNA ratio constants of donkey meat and duck meat, it detected the real donkey meat samples and the simulated samples adulterated with different proportions of duck meat (20%, 40%, 60%, and 80%, respectively). The results showed that the method had good specificity for both donkey meat and duck meat, which could be clearly distinguished from meat from horses, pigs, goats, sika deer, cattle, chickens, and dogs. The detection limit of donkey-derived DNA components was 0.01 ng/μL, and that of duck-derived DNA components was 0.1 ng/μL. Its sensitivity to duck meat components in the mixture of donkey meat and duck meat was 0.1% (w/w). The established standard curve showed a sound linear relationship. The amplification standard curve of donkey DNA was: y=−3.584x+27.003, R2=0.9982. The amplification standard curve of duck DNA was: y=−3.538x+30.907, R2=0.9991. A market pilot survey was conducted on 35 donkey meat samples using the established method, and revealed that 6 donkey meat samples (17.1%) contained duck meat. These results indicated that the real-time PCR method could be used for the rapid and accurate detection of other meat (such as duck meat) adulterated in donkey meat products. The combination of a simple detection procedure with a portable real-time PCR system provides technical support for food adulteration monitoring and control at the retail or market level.

     

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