Bellagamba Federica, Valfrè Franco, Panseri Sara, Moretti Vittorio M
Dipartimento di Scienze e Tecnologie Veterinarie per la Sicurezza Alimentare, Facoltà di Medicina Veterinaria, Università degli Studi di Milano, Via Trentacoste, 2, 20134 Milano, Italy.
J Food Prot. 2003 Apr;66(4):682-5. doi: 10.4315/0362-028x-66.4.682.
The recent European bovine spongiform encephalopathy crisis has focused attention on the importance of adopting stringent control measures to avoid the risk of the diffusion of mad cow disease through meat meal-based animal feedstuffs. Potential adulteration of such feedstuffs with bone particles from terrestrial animals is determined by microscopic examination by law before the release of these feedstuffs for free circulation in the European Community. This study describes a DNA monitoring method to examine fish meal for contamination with mammalian and poultry products. A polymerase chain reaction (PCR) method based on the nucleotide sequence variation in the 12S ribosomal RNA gene of mitochondrial DNA was developed and evaluated. Three species-specific primer pairs were designed for the identification of ruminant, pig, and poultry DNA. The specificity of the primers used in the PCR was tested by comparison with DNA samples for several vertebrate species and confirmed. The PCR specifically detected mammalian and poultry adulteration in fish meals containing 0.125% beef, 0.125% sheep, 0.125% pig, 0.125% chicken, and 0.5% goat. A multiplex PCR assay for ruminant and pig adulteration was optimized and had a detection limit of 0.25%.
近期欧洲的牛海绵状脑病危机使人们将注意力集中在采取严格控制措施以避免疯牛病通过以肉骨粉为基础的动物饲料扩散的重要性上。在这些饲料在欧洲共同体自由流通之前，依法通过显微镜检查来确定此类饲料是否可能被陆生动物的骨颗粒掺假。本研究描述了一种DNA监测方法，用于检测鱼粉是否被哺乳动物和禽类产品污染。开发并评估了一种基于线粒体DNA的12S核糖体RNA基因核苷酸序列变异的聚合酶链反应（PCR）方法。设计了三对物种特异性引物，用于鉴定反刍动物、猪和禽类的DNA。通过与几种脊椎动物物种的DNA样本进行比较，测试并确认了PCR中所用引物的特异性。该PCR能特异性检测出含有0.125%牛肉、0.125%绵羊、0.125%猪、0.125%鸡肉和0.5%山羊的鱼粉中的哺乳动物和禽类掺假情况。优化了一种用于检测反刍动物和猪掺假的多重PCR检测方法，其检测限为0.25%。
