Establishment and validation of ARMS (amplification-refractory mutation system) for identification of Anisakis species collected from Korean waters

The identification of Anisakis species in Korean waters was performed using an amplification-refractory mutation system (ARMS). ARMS is typically used to detect single nucleotide polymorphisms or allele types in the same species. However, the application of ARMS for species identification has not been reported. We designed a tetra-primer binding the internal transcribed spacer (ITS) region of 76 samples of Anisakis species and obtained reproducible results. ARMS revealed 380 bp and 130 bp ITS fragments in A. pegreffii, 380 bp and 280 bp fragments in A. simplex, a 130 bp fragments in A. typica and 380 bp, 280 bp and 130 bp fragments in an A. simplex – A. pegreffii hybrid. ARMS using a tetra-primer could be a more rapid, efficient, and reliable tool for monitoring Anisakis species more rapidly than restriction fragment length polymorphism.