Xu Jia, Zhao Wei, Zhu Mengru, Wen Yuanju, Xie Tao, He Xiaoqian, Zhang Yongfeng, Cao Suizhong, Niu Lili, Zhang Hongping, Zhong Tao
a College of Animal Science and Technology, Sichuan Agricultural University , Ya'an , Sichuan , China .
b College of Veterinary Medicine, Sichuan Agricultural University , Ya'an , Sichuan , China , and.
Mitochondrial DNA A DNA Mapp Seq Anal. 2016;27(1):628-32. doi: 10.3109/19401736.2014.908377. Epub 2014 Apr 16.
The aim of this study is to set up a protocol for identification of the adulteration in mutton based on mitochondrial 16S rRNA gene. The multiplex polymerase chain reaction (multi-PCR) assay was carried out to trace the impure DNA in mutton. A universal primer pair yielded an approximate 610 bp fragment in mutton, pork, duck, chicken, horse and cat meats. The amplicons of multi-PCR assay represented the species-specific products, which could be discriminated by the size ranging from 106 bp to 532 bp. Subsequently, the authentication of each fragment was also confirmed by sequencing. Random analyses of adulterants with various meats yielded the identical results to their components, showing the suitability of the multi-PCR assay for tracing of adulterant meats with high-accuracy and precision. This assay was sensitive to detect the species-specific DNA in different proportional mixtures of mutton and duck/pork (9.1%-90.9%). In conclusion, this multi-PCR assay successfully discriminated the double-, triple-, quadruple-, and quintuple-mixtures containing variant counterparts. This method will be particularly useful in the detection of mutton adulteration in processed foods further.
本研究的目的是建立一种基于线粒体16S rRNA基因鉴定羊肉掺假的方法。采用多重聚合酶链反应（多重PCR）检测法来追踪羊肉中不纯的DNA。一对通用引物在羊肉、猪肉、鸭肉、鸡肉、马肉和猫肉中产生了一个约610 bp的片段。多重PCR检测法的扩增产物代表物种特异性产物，可通过106 bp至532 bp的大小进行区分。随后，每个片段的鉴定也通过测序得到了证实。对各种肉类掺假物的随机分析得到了与它们的成分相同的结果，表明多重PCR检测法适用于高精度和高准确性地追踪掺假肉类。该检测法对检测羊肉与鸭肉/猪肉不同比例混合物（9.1%-90.9%）中的物种特异性DNA很敏感。总之，这种多重PCR检测法成功地区分了含有不同对应物的双重、三重、四重和五重混合物。该方法在进一步检测加工食品中的羊肉掺假方面将特别有用。
