应用荧光聚合酶链式反应检测冷冻羊肉卷中羊肉含量.docVIP

应用荧光聚合酶链式反应检测冷冻羊肉卷中羊肉含量 　　摘 要：目的：探索应用荧光聚合酶链式反应（polymerase chain reaction，PCR）法检测冷冻羊肉卷中羊肉含量。方法：以羊线粒体细胞色素B基因为羊特异性基因，线粒体12S rRNA为内参基因，应用ΔCt法对羊肉含量进行相对定量，并与称重法进行比较，分析基因拷贝数和羊肉脂肪含量对定量结果的影响。结果：荧光PCR法建立的标准曲线在羊肉含量为1%～100%时具有良好的线性关系（R2=0.990 1）。检测7 个市售冷冻羊肉卷样品，荧光PCR方法检测出的羊肉含量与称重法羊肉含量的相关系数为0.945 0（P0.001），平均绝对差异为-11.0%，95%可信区间为（-7.4%，-14.6%）。基因拷贝数不同和羊肉脂肪含量不同对内参基因扩增Ct值影响没有统计学意义（P值分别为0.072和0.206）。结论：荧光PCR法可用于快速检测冷冻羊肉卷中羊肉成分的相对定量。 　　关键词：荧光定量聚合酶链式反应；定量检测；羊肉 　　Application of Real-Time Fluorescence Polymerase Chain Reaction （PCR） for Quantitative Detection of 　　Lamb in Frozen Lamb Rolls 　　GU Wenjia， XU Qiong， ZHANG Yinan， ZHANG Qingping， QU Qinfeng 　　（Shanghai Institute of Quality Inspection and Technical Research， Shanghai 200233， China） 　　Abstract： Objective： To establish a real-time PCR method for quantitative detection of lamb in frozen lamb rolls. Methods： The ovine mitochondrial cytochrome b gene （Cyt b） as species-specific primer and the mitochondrial 12S rRNA as endogenous control were selected to conduct relative quantification through ΔCt vs. lamb content （%）. The results obtained were compared with those from the weighing method. Some factors that may affect the quantitative results including the number of gene copies and fat content in lamb were also analyzed. Results： A high correlation coefficient （R2 = 0.990 1） was showed between ΔCt and lamb content in the range of 1%?C100%. The correlation between the results of real-time PCR and weighting method for seven marketed samples of frozen lamb rolls was statistically significant （R2 = 0.945， P 0.001）， and the average absolute difference was ?11.0% （with a 95% confidence interval of ?7.4% to ?14.6%）. The effects of the number of gene copies and fat content in lamb on the quantitative detection were not statistically significant （P = 0.072， and P = 0.206， respectively）. Conclusion： The real-time PCR method can be used as a quick and practical method in quantitative detection of lamb in frozen lamb rolls. 　　Key words：