中国畜牧兽医

• 生物技术 • 上一篇    下一篇

肉制品中牛源性成分PCR检测方法的建立及其应用

熊蕊,郭凤柳,刘晓慧,赵同欣,王娜,颜红   

  1. (保定出入境检验检疫局,河北保定  071051

  • 收稿日期:2013-11-15 出版日期:2014-05-20 发布日期:2014-06-25
  • 通讯作者: 刘晓慧。E-mail:drlxh@163.com
  • 作者简介:熊蕊(1982—),女,安徽人,硕士,中级兽医师,研究方向:动物分子病毒与免疫学。
  • 基金资助:

    河北出入境检验检疫局自主立项项目(HE2011K019)。

Establishment and Application of Method for PCR Detection of Beef Ingredient in Meat Products

XIONG Rui,GUO Feng-liu,LIU Xiao-hui,ZHAO Tong-xin,WANG Na,YAN Hong   

  1. Baoding Entry-exit Inspection and Quarantine BureauBaoding 071051China

  • Received:2013-11-15 Online:2014-05-20 Published:2014-06-25

摘要: 根据牛特异性线粒体DNA片段,设计合成1对引物,以生、熟牛肉为材料,建立了肉制品中牛源性成分的PCR检测方法,并用该法对市售的67份牛肉制品进行检测。结果显示,所检牛源性成分在271 bp处出现预期的条带,扩增片段经Sau3AⅠ酶切分析确认,获得的214和57 bp片段与预期一致;运用该引物均可扩增出水牛肉、牦牛肉、奶牛肉、黄牛肉单一的相同大小的DNA条带,而对羊、马、狗、驴、兔和鸭等14种动物肉的DNA扩增则呈阴性,其检测灵敏度达到53.2 fg/μL DNA;利用该法对67份牛肉制品进行检测,检出率为100%。结果表明,该法快速简便,且具有较高的特异性和敏感性,可用于市售牛肉制品中牛源性成分的鉴定。

关键词: PCR方法; 检测; 牛源性成分; 特异性; 灵敏度

Abstract: This study established a PCR method for detection the beef ingredient in meat products according to the bovine specific mitochondrial DNA fragment,and 67 meat products of cow were detected using PCR.The results showed that a specific amplification fragment of the expected size was detected and demonstrated,buffalo,yak,cow and bovine meat were positive using this method,whereas the 14 kinds of animal meat such as sheep, equine, canis, donkey, rabbit and duck were negative,the detection limit was 53.2 fg/μL DNA.The positive rate of 67 meat products of cow was 100%.The results indicated that the method was quickness,convenience,sensitivity and specificity,and can be used for the identification of bovine source of beef products components.

Key words: PCR method; detection; beef ingredient; specificity; sensitivity