肝水解肽样品中牛源及猪源性成分的PCR检测

Identification of bovine and porcine derived materials in liver hydrolysate samples by PCR

目的:建立肝水解肽各工艺步骤段中样品的PCR检测方法,用于检测生产原料的动物来源是牛源性或猪源性。方法:分别对猪源性及牛源性肝水解肽各工艺步骤段中样品进行了DNA提取,并加入猪、牛特异性引物进行扩增。对扩增产物进行酶切及测序验证。结果:工艺段为超滤液四之前的样品均可提取并扩增出相应的DNA片段,牛源性PCR产物与Genbank中牛序列的同源性为100%,猪源性PCR产物与猪序列的同源性为99.4%,PCR扩增的检测限为0.5 mg·mL-1的肝细胞酶解液。结论:可用本方法检测肝水解肽各工艺步骤段中超滤液四之前的样品。

Objective：To develop a PCR method for identification bovine or porcine derived materials in liver hy-drolysate samples.Methods：DNA were extracted from liver hydrolysate samples which prepared by different steps of the preparation process,and then they were amplified with specific primers.PCR products were digested by enzyme and sequenced to verify the sequence identity.Results：Characteristic DNA fragments were extracted and amplified in the samples prepared in the process steps before the fourth ultrafiltrate.It was found that PCR products amplified from bovine-derived samples and porcine-derived samples have 100% and 99.4% homologies to that of gene in the Genbank respectively.The method was confirmed with a minimum detection level of 0.5 mg·mL-1 liver cell hydrolysate.Conclusion：The method can be applied to detect the DNA fragments in the samples prepared in the process before the fourth ultrafiltrate.