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

Abstract

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\u202fbp and 130\u202fbp ITS fragments in A. pegreffii, 380\u202fbp and 280\u202fbp fragments in A. simplex, a 130\u202fbp fragments in A. typica and 380\u202fbp, 280\u202fbp and 130\u202fbp 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.