实时荧光PCR法检测肉制品中猪源性成分

目的S建立一个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S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300S,30S,3S,0.3S和0.03Sng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S以Ct值作为纵坐标,以lgXS(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,SR2=0.9997；该方法的检测灵敏度达到2 Spg。结论S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。

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

Objective To establish a highly specific and sensitive method for the detection of swine derived ingredients. Methods A pair of specific primers were designed and synthesized based on porcine estrogen receptor gene. DNA was extracted from pork by liquid nitrogen grinding combined with kit extraction. Gradient reaction was performed with 300, 30, 3, 0.3 and 0.03 ng as templates respectively. The detection method was established based on fluorescent quantitative PCR (SYBR). 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 50 pg. Conclusion The method is simple, rapid, specific, reproducible and sensitive. It can be used as a feasible method for market supervision and identification.