PCR-RFLP pattern of three kinds of Metagonimus in Korea

We tried to compare the three kinds of Metagonimus species, M. yokogawai, Metagonimus Miyata type, and M. takahashii, which were known to be distributed in Korea with polymerase chain reaction based-restriction fragment length polymorphism (PCR-RFLP) patterns. We amplified the internal transcribed spacer 1 (ITS1) site of ribosomal RNA and mitochondrial cytochrome c oxidase I (mCOI) gene. The restriction patterns of ITS1 gene with Rsa I, Alu I and Msp I showed multiple fragmented bands of different sizes between three kinds of Metagonimus. In case of mCOI gene, Rsa I and Alu I enzymes produced differentially fragmented band patterns. According to the parsimony analysis of PCR-RFLP patterns, the estimated genetic divergence between M. yokogawai and Metagonimus Miyata type was 0.034880, between Metagonimus Miyata type and M. takahashii was 0.028098, between M. yokogawai and M. takahashii was 0.018179. It is suggested that Metagonimus Miyata type may be separate species and evolutionize at the older time than the other two species.