Matsumoto Chihiro, Kyota Yukino, Yamanaka Shunya, Murakawa Naoki, Kikunaga Ryutaro, Yamada Yoshihiro, Kawachi Hiroyuki
Faculty of Bioscience, Nagahama Institute of Bio-Science and Technology, Tamura 1266, Nagahama, Shiga 526-0829 Japan.
J Food Sci Technol. 2019 Oct;56(10):4705-4713. doi: 10.1007/s13197-019-03914-3. Epub 2019 Jul 5.
Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis was used to identify meat from biwa trout (), amago trout (), yamame trout (), and rainbow trout (). PCR amplification was conducted using primers flanking conserved regions of NADH dehydrogenase subunits 4 and 5 (ND4-ND5) (2848 bp) and ND1 (1091 bp) genes of mitochondrial DNA following restriction digestion with the enzyme III. Although the segments of ND4-ND5 and ND1 genes showed intraspecies variation, the generation of DNA fragments larger than 300 bp and 160 bp following cleavage by HaeIII of ND4-ND5 and ND1, respectively, was efficient to differentiate the four species. Furthermore, this method was successful in species identification even when using PCR-amplified products obtained from thermally processed biwa trout samples. This sensitive technique can be utilized to reveal commercial fraud, where biwa trout is adulterated with meat from cheaper counterparts.
聚合酶链反应-限制性片段长度多态性（PCR-RFLP）分析用于鉴定琵琶湖鳟、阿马戈鳟、山女鳟和虹鳟的鱼肉。使用位于线粒体DNA的烟酰胺腺嘌呤二核苷酸脱氢酶亚基4和5（ND4-ND5）（2848 bp）以及ND1（1091 bp）基因保守区域两侧的引物进行PCR扩增，随后用酶III进行限制性消化。尽管ND4-ND5和ND1基因片段表现出种内变异，但分别用HaeIII切割ND4-ND5和ND1后产生大于300 bp和160 bp的DNA片段，能有效区分这四个物种。此外，即使使用从热处理的琵琶湖鳟样品中获得的PCR扩增产物，该方法在物种鉴定中也取得了成功。这种灵敏的技术可用于揭示商业欺诈行为，即琵琶湖鳟肉被掺假了更便宜的同类鱼肉。
