Fajardo Violeta, González Isabel, Martín Irene, Rojas María, Hernández Pablo E, García Teresa, Martín Rosario
Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense, 28040 Madrid, Spain.
Meat Sci. 2008 Jun;79(2):289-98. doi: 10.1016/j.meatsci.2007.09.013. Epub 2007 Oct 5.
A rapid real-time polymerase chain reaction (PCR) technique using SYBR Green detection system, has been developed for the quantification of red deer, fallow deer, and roe deer DNAs in meat mixtures. The method combines the use of cervid-specific primers that amplify a 134, 169, and 120bp of the 12S rRNA gene fragment of red deer, fallow deer and roe deer, respectively, and universal primers that amplify a 140bp fragment on the nuclear 18S rRNA gene from eukaryotic DNA. The C(t) (threshold cycle) values obtained with the 18S rRNA primers are used to normalize those obtained from each of the cervid-specific systems, serving as endogenous control for the total content of PCR-amplifiable DNA in the sample. Analysis of experimental raw and heat treated binary mixtures of red deer, fallow deer or roe deer meat in a swine meat matrix demonstrated the suitability of the assay for the detection and quantification of the target cervid DNAs in the range 0.1-0.8%, depending on the species and treatment of the meat samples analyzed.
一种使用SYBR Green检测系统的快速实时聚合酶链反应（PCR）技术已被开发出来，用于定量检测混合肉类中马鹿、黇鹿和狍子的DNA。该方法结合使用了鹿科特异性引物和通用引物，其中鹿科特异性引物分别扩增马鹿、黇鹿和狍子12S rRNA基因片段的134bp、169bp和120bp，通用引物则从真核DNA的核18S rRNA基因上扩增出一个140bp的片段。用18S rRNA引物获得的C(t)（阈值循环）值用于对从每个鹿科特异性系统获得的值进行标准化，作为样品中可PCR扩增DNA总含量的内参。对猪肉基质中马鹿、黇鹿或狍子肉的实验性生肉和热处理二元混合物的分析表明，根据所分析肉样的种类和处理方式，该检测方法适用于检测和定量0.1%-0.8%范围内的目标鹿科DNA。
