聚合酶链反应(PCR)鉴定猪肉方法的建立及应用

Establishment and Application of a PCR for the Identification of Pork

依据猪小卫星ＤＮＡ序列设计了１对引物，建立了ＰＣＲ鉴定生、熟猪肉的方法。应用本方法特异地扩增出预期的猪３０７ｂｐ的ＤＮＡ片段，其检测敏感度对生猪肉达３８．６ｆｇＤＮＡ，对熟猪肉和高压猪肉为０．３７５ｆｇ全基因组ＤＮＡ。运用该引物仅可扩增生猪ＤＮＡ的单一片段，而对马、牛、羊等１２种动物肉的ＤＮＡ扩增则呈阴性。应用本法对６３份生、熟猪肉进行鉴定，正确率为１００％，对各种样品检测，均可在６ｈ内完成。

A polymerase chain reaction (PCR)based method for the identification of swine meat by amplification of swine microsatellite DNA was established.The method not was only able to amplify raw swine DNA,but also cooked or autoclaved meat DNA.The sequence selected for amplification consisted of a 307 bp DNA fragment lying in the microsatellite DNA of swine.The specificity test demonstrated that this method was positive for raw swine meat, cooked and auto claved swine meat DNA ,but negative for equine,sheep,bovine,camel meat DNA .At least 38.6 fg of DNA from raw swine samples and 0.375 pg of DNA from cooked or autoclaved swine meat samples were detected respectively by PCR. We tested 63 swine samples by PCR and found a rate of correct identification up to 100%.The method needed only 6h for detection of meat products of all kinds.The results showed that the PCR method was sensitive,specific,convenient and rapid,so it may be suitable for rapid identification of swine meat.