Matsunaga T, Chikuni K, Tanabe R, Muroya S, Shibata K, Yamada J, Shinmura Y
Japan Meat Processors Association, Ebisu 1-5-6 Shibuya-ku, Tokyo 150, Japan.
Meat Sci. 1999 Feb;51(2):143-8. doi: 10.1016/s0309-1740(98)00112-0.
The polymerase chain reaction (PCR) was applied to identify six meats (cattle, pig, chicken, sheep, goat and horse) as raw materials for products. By mixing seven primers in appropriate ratios, species-specific DNA fragments could be identified by only one multiplex PCR. A forward primer was designed on a conserved DNA sequence in the mitochondrial cytochrome b gene, and reverse primers on species-specific DNA sequences for each species. PCR primers were designed to give different length fragments from the six meats. The products showed species-specific DNA fragments of 157, 227, 274, 331, 398 and 439 bp from goat, chicken, cattle, sheep, pig and horse meats, respectively. Identification is possible by electrophoresis of PCR products. Cattle, pig, chicken, sheep and goat fragments were amplified from cooked meat heated at 100 or 120°C for 30 min, but horse DNA fragments could not be detected from the 120°C sample. Detection limits of the DNA samples were 0.25 ng for all meats.
采用聚合酶链反应(PCR)来鉴定六种肉类(牛、猪、鸡、羊、山羊和马)作为产品的原材料。通过以适当比例混合七种引物,仅通过一次多重PCR就能鉴定出物种特异性DNA片段。在细胞色素b线粒体基因的保守DNA序列上设计正向引物,在每个物种的物种特异性DNA序列上设计反向引物。PCR引物设计为从六种肉类中产生不同长度的片段。产物分别显示出来自山羊、鸡、牛、羊、猪和马肉的157、227、274、331、398和439 bp的物种特异性DNA片段。通过PCR产物的电泳可以进行鉴定。牛、猪、鸡、羊和山羊的片段可从在100或120°C加热30分钟的熟肉中扩增出来,但在120°C的样品中未检测到马的DNA片段。所有肉类DNA样品的检测限均为0.25 ng。