1Department of Materials and Life Science, Faculty of Science and Technology, Shizuoka Institute of Science and Technology, Fukuroi, Shizuoka 437-8555, 2Hikari Corporation Co. Ltd., Iwata, Shizuoka 438-0051, 3Miyabi Suisan Co. Ltd., Suruga, Shizuoka 422-8054, 4Atsumi Co. Ltd., Toyohashi, Aichi 441-8041, Japan
A DNA analysis method has been developed for detecting the mixing of foreign species of eel among about 100 putative all Anguilla japonica eels, with high accuracy and low cost at the level of commercial products. Allele specific PCR (AS-PCR) assay was used to detect the non-A. japonica allele by using primers that match the 3′ end nucleotides of the mitochondrial DNA cytochrome b gene sequence of all known eel species except A. japonica. Foreign species specific PCR products, if obtained, were subjected to restriction fragment length polymorphism (RFLP) assay to quickly identify the species from the difference of digestion fragment patterns by using Rsa I and Hinf I as the restriction enzymes.