Rodríguez Miguel A, García Teresa, González Isabel, Asensio Luis, Hernández Pablo E, Martín Rosario
Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense, 28040 Madrid, Spain.
J Agric Food Chem. 2004 Mar 24;52(6):1478-83. doi: 10.1021/jf035240n.
A real-time quantitative Polymerase Chain Reaction (PCR) method has been developed for the quantitation of mule duck (Anas platyrhynchos x Cairina moschata) in binary duck/goose foie gras mixtures. The method combines the use of real-time PCR with duck-specific and endogenous control "duck + goose" primers to measure duck content and total foie gras content, respectively. Both PCR systems (duck-specific and duck + goose) were designed on the mitochondrial 12S ribosomal RNA gene (rRNA). The duck-specific system amplifies a 96 bp fragment from duck DNA, whereas the duck + goose system amplifies a 120 bp fragment from duck and goose DNA. The method measures PCR product accumulation through a FAM-labeled fluorogenic probe (TaqMan). The C(t) (threshold cycle) values obtained from the duck + goose system are used to normalize the ones obtained from the duck-specific system. Analysis of experimental duck/goose foie gras binary mixtures demonstrated the suitability of the assay for the detection and quantitation of duck in the range of 1-25%. This genetic marker can be very useful to avoid mislabeling or fraudulent species substitution of goose by duck in foie gras.
已开发出一种实时定量聚合酶链反应（PCR）方法，用于定量二元鸭/鹅肥肝混合物中的骡鸭（绿头鸭×疣鼻栖鸭）。该方法将实时PCR与鸭特异性及内参“鸭+鹅”引物结合使用，分别测量鸭含量和总肥肝含量。两种PCR系统（鸭特异性和鸭+鹅）均基于线粒体12S核糖体RNA基因（rRNA）设计。鸭特异性系统从鸭DNA中扩增出一个96 bp的片段，而鸭+鹅系统从鸭和鹅DNA中扩增出一个120 bp的片段。该方法通过FAM标记的荧光探针（TaqMan）测量PCR产物的积累。从鸭+鹅系统获得的C(t)（阈值循环）值用于对从鸭特异性系统获得的值进行标准化。对实验性鸭/鹅肥肝二元混合物的分析表明，该检测方法适用于检测和定量1%-25%范围内的鸭。这种遗传标记对于避免肥肝中鸭误标或用鸭欺诈性替代鹅的物种非常有用。
