肉制品中牛源性成分PCR检测方法的建立及其应用

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

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271bp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57bp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53.2fg/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。

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.