Kesmen Z, Gulluce A, Sahin F, Yetim H
Erciyes University, College of Engineering, Food Engineering Department, 38039 Kayseri, Turkey.
Meat Sci. 2009 Aug;82(4):444-9. doi: 10.1016/j.meatsci.2009.02.019. Epub 2009 Mar 20.
In this study, a convenient, sensitive and specific real-time PCR assay was described for the species identification and their quantification in raw and cooked meat products. Specific primers and TaqMan probes were designed on the mitochondrial ND2, ND5 and ATP 6-8 genes for donkey, pork and horse, respectively, and the performance of the method was tested. In the results, no cross-reaction was observed between the donkey and pork species specific primer-probe systems and non-target species (bovine, ovine, chicken and turkey). Only one cross reaction was observed between the horse species specific primer-probe set and 100ng pork DNA at the ct 33.01 level (corresponding to 0.01ng horse DNA). The real-time quantitative assay used in this study allowed the detection of as little as 0.0001ng template DNA from pure meat for each species investigated and experimental meat mixtures. In conclusion, it can be suggested that the TaqMan probe assay used in this research might be a rapid and sensitive method for the routine meat species identifications studies in raw or cooked meat products.
在本研究中，描述了一种简便、灵敏且特异的实时荧光定量PCR检测方法，用于生肉和熟肉产品中物种的鉴定及其定量分析。分别针对驴、猪和马的线粒体ND2、ND5和ATP 6 - 8基因设计了特异性引物和TaqMan探针，并对该方法的性能进行了测试。结果显示，驴和猪的物种特异性引物-探针系统与非目标物种（牛、羊、鸡和火鸡）之间未观察到交叉反应。在ct值为33.01水平（相当于0.01ng马DNA）时，仅在马的物种特异性引物-探针组与100ng猪DNA之间观察到一次交叉反应。本研究中使用的实时定量检测方法能够检测出所研究的每个物种纯肉以及实验性肉混合物中低至0.0001ng的模板DNA。总之，可以认为本研究中使用的TaqMan探针检测法可能是一种用于生肉或熟肉产品中常规肉类物种鉴定研究的快速且灵敏的方法。
