实时荧光PCR法检测肉制品中猪源性成分

Detection of porcine-derived components in meat products by real-time fluorescence PCR

中文关键词:  猪源性成分  实时荧光PCR  肉类制品

英文关键词:porcine-derived ingredients  real-time fluorescent PCR  meat product

中文摘要:

      目的 建立一种高特异性和高灵敏度的有效检测猪源性成分的检测方法。方法 以猪雌激素受体基因设计合成一对特异性引物, 采用液氮研磨结合试剂盒抽提的方法提取猪肉中的DNA, 分别以300、30、3、0.3、0.03 ng为模板量进行梯度反应, 基于荧光定量PCR技术(SYBR法)建立检测方法。结果 以Ct值作为纵坐标, 以lgX(X为DNA的量, ng)的值作为横坐标, 得到标准曲线方法Ct=30.473?3.542lgX, r2=0.9997; 该方法的检测灵敏度达到2 pg。结论 该猪源性成分检测方法简单快捷、特异性和重复性好、灵敏度高, 可作为市场监督和检测鉴定的可行性方法。

英文摘要:

      Objective To establish a detection method with high specificity and high sensitivity for effectively detecting porcine-derived components. Methods A pair of specific primers were designed and synthesized based on porcine estrogen receptor gene. DNA in pork was extracted by liquid nitrogen milling combined with kit extraction. Gradient reaction was performed with 300, 30, 3, 0.3 and 0.03 ng as the template amount, respectively. The detection method was established based on fluorescent quantitative PCR (SYBR method). Results Using Ct value as ordinate and lgX (X as DNA quality, ng) as abscissa, the standard curve method was obtained, Ct=30.473?3.542lgX, r2= 0.9997. The detection sensitivity of this method was 2 pg. Conclusion The detection method for the porcine-derived components is simple and rapid, good in specificity and repeatability, and high in sensitivity, and can be used as a feasible method for market supervision, detection and identification.