四重PCR反应体系的建立及在牛肉掺假检测中的应用  

Establishment of Quadruple PCR Reaction System:Its Application in Detection of Adulterated Beef

作　　者：钟文涛[1,2] 王芳妹[1,2] 李白玉 蒋伟 周丛 

机构地区：[1]湖南省产商品质量监督检验研究院(国家农副产品质量监督检验中心),湖南长沙410007 [2]湖南师范大学生命科学院,湖南长沙410006

出　　处：《现代生物医学进展》2016年第18期3574-3577,共4页Progress in Modern Biomedicine

基　　金：湖南省2012年第六批省级科技计划项目(2012SK3290)

摘　　要：目的:建立马、牛、猪、鸭肉四重聚合酶链式反应(PCR)反应体系,为牛肉掺假检测提供参考。方法:选取马、牛、猪、鸭肉阳性样本,根据线粒体基因组中细胞色素b(cytochrome b)序列,设计多重PCR引物,同时对马、牛、猪、鸭肉进行PCR及电泳检测,建立四重PCR反应体系,并运用该体系对市场40份牛肉制品进行检测。结果:引物比例为马:牛:猪:鸭=1:1:2:1时,46℃退火温度下反应35个循环,四重PCR效果较好;建立四重PCR反应体系,四重PCR条带清晰且亮度较为一致,几乎没有二聚体产生,扩增灵敏度高。根据样品检测结果,烤牛肉串、牛肉丸的掺假率较高,分别为36.4%、75%,牛排未检出除牛肉外的三种动物源成分,但不排除有其他动物源成分的掺入。结论:四重PCR反应体系可用于牛肉掺假检测。Objective： To establish the quadruple PCR reaction system of horsemeat, beef, pork and duck,in order to provide a reference for the detection of adulterated beef. Methods： The positive samples of horsemeat, beef, pork and duck were selected. The multiplex PCR primers were designed according to the sequence of cytochrome b in the mitochondrial genome; the horsemeat, beef, pork and duck were detected by PCR and electrophoresis test at the same time; the quadruple PCR reaction system was established, then the system was used to detect 40 beef products in the market. Results： The effect of quadruple PCR was better when the ratios of primer of horsemeat were beef： pork： duck=1：1：2：1, and 35 cycles reacted under the 46℃ annealing temperature. The quadruple PCR reaction system was established, the strip was clear and bright, almost no dimer was generated, and the amplified sensitivity was high. The beef products detection results showed that the adulteration rate of roast beef string,stewed beef meatballs were higher, which were 36.4% and75% respectively, while the beefsteak had no other three kinds of animal ingredients except beef,but can not rule out the incorporation of other animal ingredients. Conclusion： The quadruple PCR reaction system can be applied to detecting adulterated beef.

关 键 词：四重PCR 细胞色素B 牛肉掺假 检测 

分 类 号：R155.62[医药卫生—营养与食品卫生学] S858[医药卫生—公共卫生与预防医学]