同时鉴定9种肉源种属的复合检测体系的建立及其应用

Establishment and application of a composite detection system for simultaneous identification of 9 meat species

目的建立一种可以同时鉴定猪、牛、羊、鸡、鸭、猫、狗、鼠和鲤鱼的多重PCR检测方法,并测试其技术性能指标,评估其在法医学或食品安全事件中的应用价值。方法根据上述9种动物的线粒体细胞色素b基因,利用其片段中种间特异性强的序列设计引物,采用毛细管电泳检测平台对各种属PCR扩增产物进行检测,依据扩增片段长度的差异与标记的荧光对初始模板的肉源种属进行鉴别;并通过扩增特异性、实际样本测试、混合样本检测灵敏度等指标评价该方法在实际法医学或食品安全案件中的应用价值。结果经过验证,建立的同时鉴定9种肉源种属的复合检测体系对其中任意一个种属的检测特异性均较高,且灵敏度较强,能够满足绝大多数法医学或食品安全案件中的检测要求。结论本研究建立的肉源种属复合检测体系可以在法医学未知种属样本的鉴定及食品安全肉类掺假等案件中提供帮助。

Objective To establish a multiplex PCR method for simultaneous identification of pigs, cattle, sheep, chickens, ducks, cats, dogs, rats and carps, test its technical performance indicators and evaluate its application value in forensic science or food safety incidents. Methods Based on the mitochondrial cytochrome b gene of the above 9 species of animals the primers were designed by using the strong interspecific sequence of the fragments. Capillary electrophoresis detection platform was used to detect PCR amplification products, and according to the difference of the amplified fragment length and the fluorescence of the marker, the meat derived biomaterials of the original template was identified. The application value of this method in practical forensic medicine or food safety cases was evaluated by such indicators as amplification specificity, actual sample test and mixed sample detection sensitivity. Results After verification, the established complex detection system for identifying nine meat-derived species simultaneously has high specificity for any species. Further more, the sensitivity is strong enough to meet the requirements of most forensic and food safety incidents. Conclusion The composite detection system of meat-derived species established in this study can provide assistance in the identification of unknown species in forensic science and food-safe meat adulteration.