Dalmasso A, Fontanella E, Piatti P, Civera T, Rosati S, Bottero M T
Dipartimento di Patologia Animale, Università degli Studi di Torino, Via Leonardo da Vinci 44, Grugliasco, Turin 10095, Italy.
Mol Cell Probes. 2004 Apr;18(2):81-7. doi: 10.1016/j.mcp.2003.09.006.
A multiplex Polymerase Chain Reaction (PCR) assay was applied to feedstuff analysis for the identification of the most used species in rendering plants (ruminant, poultry, fish and pork materials). Primers were designed in different regions of mitochondrial DNA (12S rRNA, tRNA Val and 16S rRNA) after alignment of the available sequences in the GenBank database. The primers generated specific fragments of 104-106, 183, 220-230 and 290 bp length for ruminants, poultry, fish and pork, respectively. The detection limit was 0.004% for fish primers and 0.002% for ruminants, poultry and pork primers. The multiplex PCR proposed in this study can be considered a valid alternative to the microscopic method for the detection of animal derived materials banned by a European Union Regulation as a preventive measure against the spread of Bovine Spongiform Encephalopathy.
采用多重聚合酶链反应（PCR）分析法对饲料进行分析，以鉴定在动物饲料加工厂中最常用的物种（反刍动物、家禽、鱼类和猪肉原料）。在对GenBank数据库中可用序列进行比对后，在线粒体DNA的不同区域（12S rRNA、tRNA Val和16S rRNA）设计了引物。这些引物分别为反刍动物、家禽、鱼类和猪肉生成了长度为104 - 106、183、220 - 230和290 bp的特异性片段。鱼类引物的检测限为0.004%，反刍动物、家禽和猪肉引物的检测限为0.002%。本研究中提出的多重PCR可被视为一种有效的替代方法，用于替代显微镜检测法，以检测欧盟法规禁止的动物源性材料，作为预防牛海绵状脑病传播的一项措施。
