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| 鉴定动物奶源的多重RCR方法建立及应用 |
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王天添1,刘悦1,赵远1,王艳双2,李明成1,3,孙丽媛1*
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1.北华大学 医学技术学院, 吉林 吉林132013;2.北华大学 医学院, 吉林 吉林 132013;3.吉林省中药DNA指纹检测技术科技创新中心, 吉林 吉林132013
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| 摘要: |
| 为准确鉴别牛奶、山羊奶、绵羊奶、马奶以及骆驼奶，利用多重PCR方法建立奶制品DNA提取方法及奶制品质量检测体系。结果表明: 1)建立的奶制品DNA提取方法不受食品添加剂影响，提取过程简便快速高效，可于2 h之内全部完成，提取质量浓度不低于100 ng/μL，纯度介于1.70~2.10，适用于常见液态奶DNA提取；2)建立的多重PCR体系中各引物种属特异性强，无交叉干扰；3)检测灵敏度为1 ng/μL；4)对24 份人工模拟混合奶样进行检测，准确度达100%，随机抽样重复检测3 次，检测结果稳定；5)对本地区市场销售液态奶样品10份(牛奶5份，羊奶1份，山羊奶1份，马奶2份，骆驼奶1份)进行检测:牛奶、马奶及骆驼奶奶源检测结果与产品标识相符；2份羊奶样品分别标识鲜羊奶及鲜山羊奶，但检测结果显示存在牛源性强阳性信号，怀疑存在牛源性成分掺假。 |
| 关键词: 奶制品 DNA提取 多重PCR 特色奶 |
| DOI：10.11841/j.issn.1007-4333.2020.11.08 |
| 分类号: |
| 基金项目:吉林省科技发展计划项目(20190301014NY,20180201023YY,20190304115YY,20190303093SF) |
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| Establishment and application of multiplex PCR method for the identification of animal milk sources |
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WANG Tiantian1,LIU Yue1,ZHAO Yuan1,WANG Yanshuang2,LI Mingcheng1,3,SUN Liyuan1*
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1.College of Laboratory Medical, Beihua University, Jilin, 132013, China;2.College of Medical, Beihua University, Jilin, 132013, China;3.Jilin Province Traditional Chinese Medicine DNA Fingerprint Detection Technology Innovation Center, Jilin, 132013, China
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| Abstract: |
| To accurately identify milk, goat milk, sheep milk, horse milk and camel milk, the multiplex PCR was used to establish dairy product DNA extraction method and quality detection system. The results indicated that: 1)The established DNA extraction method for dairy products was simple, fast and efficient, not affected by food additives. The method could be completed in 2 hours with the extraction concentration of no less than 100 ng/μL and purity between 1. 70 and 2. 10, which was suitable for the DNA extraction of common liquid milk. 2)In the established multiplex PCR system, all primers had strong species specificity and no cross interference. 3)The detection sensitivity was 1 ng/μL. 4)A total of 24 samples mixed milk samples were tested for three times, the accuracy reached 100%, and the results were stable. 5)Ten samples of liquid milk(5 milk, 1 goat milk, 2 horse milk and 1 camel milk)on the local market were tested. The results showed that the source test of milk, horse milk and camel milk were consistent with product identification. However, two samples marked as fresh sheep's milk and fresh goat's milk displayed strong positive signals of bovine origin, suggesting that there was bovine origin adulteration. |
| Key words: dairy products DNA extraction multiplex PCR specialty milk |
