生物技术通报 ›› 2020, Vol. 36 ›› Issue (1): 202-208.doi: 10.13560/j.cnki.biotech.bull.1985.2019-0607

• 技术与方法 • 上一篇    下一篇

PCR-膜芯片技术在牦牛及犏牛肉制品的真假鉴定中的应用

赵睿骁1,3, 韩斐1,3, 江明锋2,3, 曾莲4, 韩嘉钰4, 黄盈4, 周航4   

  1. 1. 西南民族大学青藏高原研究院,成都 610041;
    2. 西南民族大学生命科学与技术学院,成都 610041;
    3. 西南民族大学青藏高原动物遗传资源保护与利用重点实验室,成都 610041;
    4. 四川华汉三创生物科技有限公司,成都 610093
  • 收稿日期:2019-07-05 出版日期:2020-01-26 发布日期:2020-01-08
  • 作者简介:赵睿骁,男,硕士研究生,研究方向:分子遗传学与基因工程;E-mail:243781387@qq.com
  • 基金资助:
    西南民族大学研究生创新型项目(CX2019SZ83),西南民族大学中央高校基本科研业务费专项资金,国家级项目后期资助项目(2019XMJXPY09)

Application of PCR-membrane Chip Technology in the Identification of Yak and Yak-cattle Meat

ZHAO Rui-xiao1,3, HAN Fei1,3, JIANG Ming-feng2,3, ZENG Lian4, HAN Jia-yu4, HUANG Ying4, ZHOU Hang4   

  1. 1. Institute of Qinghai-Tibetan Plateau,Southwest Minzu University,Chengdu 610041;
    2. College of Life Science and Technology,Southwest Minzu University,Chengdu 610041;
    3. Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization,Southwest Minzu University,Chengdu 610041;
    4. Sichuan Hua Hansan Bio Technology Co.,Ltd.,Chengdu 610093
  • Received:2019-07-05 Published:2020-01-26 Online:2020-01-08

摘要: 旨在利用PCR-膜芯片技术快速高效的鉴定牦牛及犏牛肉制品的真假。采集样品(猪、山羊、黄牛、水牛、牦牛、鸡、鸭、兔、狗、真犏牛、假犏牛肉样各8个,19个肉干制品样品),提取并纯化DNA,利用11对特异性引物进行多重PCR,将扩增产物与含有12个探针的膜芯片反向点杂交,测试膜芯片的准确度及灵敏度,并鉴定市场上销售的牦牛肉制品真假。结果表明,膜芯片准确度高,特异性强,无交叉反应,灵敏度能达到0.1 ng,检测灵敏度高。通过鉴定市场上销售的假牦牛肉干制品几乎都以黄牛和水牛肉为原料制作。应用PCR-膜芯片技术可快速准确的鉴定出牦牛肉制品的真假,有利于打击假冒产品,维护市场平衡。

关键词: 牦牛, 犏牛, 多重PCR, 膜芯片

Abstract: This paper aims to quickly and efficiently identify the authenticity of yak and yak-cattle meat products by PCR-membrane chip technology. We collected samples(8 fresh meat and 19 dry meat samples of fresh pig,goat,oxen,buffalo,yak,chicken,duck,rabbit,dog,and yak-cattle),extracted and purified DNA,used 11 specific primers to perform multiplex PCR. Further we hybridized the amplified products with a membrane chip containing 12 probes via reverse dot blot,measured the accuracy and sensitivity of the membrane chip,and identified the authenticity of the commercially available yak meat products. The results demonstrated that the membrane chip was of high accuracy,strong specificity,no cross-reaction,and high detection sensitivity(low to 0.1 ng). Detection results of the commercial fake yak meat products showed that almost all of them were made from yellow cattle and buffalo meat. In conclusion,the application of PCR-membrane chip technology can quickly and accurately identify the real and fake yak meat products,which is conducive to combating counterfeit products and maintaining market balance.

Key words: yak, yak-cattle, multiplex PCR, membrane chip