Shantou Entry-Exit Inspection and Quarantine Bureau, Shantou, China.
Guangdong Entry-Exit Inspection and Quarantine Bureau, Guangzhou, China.
Meat Sci. 2018 Mar;137:41-46. doi: 10.1016/j.meatsci.2017.11.003. Epub 2017 Nov 14.
Meat adulteration incidents have been reported frequently over the past few years, and the corresponding traceability issues attracted much more attention due to the customer's demands and administration's responsibility. Therefore, it is important to develop high-throughput and rapid detection methods to identify the specific sources from meat samples. In this study, a multiplex TaqMan locked nucleic acid real-time polymerase chain reaction assay (MLNA-RT-PCR) was developed to simultaneously detect multiple meat sources (duck, pork, beef and chicken). PCR primers and TaqMan-LNA probes were designed based on species-specific mitochondrial gene sequences, and the MLNA-RT-PCR was developed and optimized for better performance. The specificity of this assay was verified through identifying unrelated (sheep, horse, deer, donkey, rabbit, goose, goat, shrimp, salmon and maize) mitochondrial DNA as species-specific targets. The detection limit for MLNA-RT-PCR reached to the level of 0.01% of each species. The assay was then used to identify the meat sources of commercial meat and meat-derived products that were obtained from markets in Shantou, and the results were 98% consistent with that obtained from detection based on the national standard. In conclusion, this MLNA-RT-PCR is a high-throughput, sensitive and specific method that can be used to identify multiple meat sources in meat and meat-derived products.
近年来，肉类掺假事件屡有报道，由于消费者的需求和管理部门的责任，相应的可追溯性问题引起了更多的关注。因此，开发高通量和快速的检测方法来识别肉类样品中的具体来源非常重要。在本研究中，开发了一种多重 TaqMan 锁核酸实时聚合酶链反应检测方法（MLNA-RT-PCR），以同时检测多种肉类来源（鸭、猪、牛和鸡）。根据种特异性线粒体基因序列设计了 PCR 引物和 TaqMan-LNA 探针，并对 MLNA-RT-PCR 进行了开发和优化，以获得更好的性能。通过鉴定与无关物种（绵羊、马、鹿、驴、兔、鹅、山羊、虾、鲑鱼和玉米）的线粒体 DNA 作为种特异性靶标，验证了该检测方法的特异性。MLNA-RT-PCR 的检测限达到了每种物种的 0.01%。然后，该检测方法被用于鉴定从汕头市市场获得的商业肉类和肉类衍生产品的肉类来源，结果与基于国家标准的检测结果 98%一致。总之，该 MLNA-RT-PCR 是一种高通量、敏感和特异性的方法，可用于识别肉类和肉类衍生产品中的多种肉类来源。
