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Korean J Parasitol > Volume 35(4):1997 > Article

Original Article
Korean J Parasitol. 1997 Dec;35(4):271-276. English.
Published online Dec 20, 1997.  http://dx.doi.org/10.3347/kjp.1997.35.4.271
Copyright © 1997 by The Korean Society for Parasitology
PCR-RFLP pattern of three kinds of Metagonimus in Korea
J R Yu,*1J S Chung,1S Huh,2S H Lee,3 and J Y Chai3
1Department of Parasitology, College of Medicine, Kon-Kuk University, Chungju, Korea.

*Corresponding author (Email: yu@kcucc.konkuk.ac.kr)
Received October 16, 1997; Accepted November 10, 1997.


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 legth 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.


Fig. 1
PCR products for the ITS1 region of rDNA of three Metagonimus spp. Lane 1, Metagonimus Miyata type; lane 2, M. yokogawai; lane 3, M. takahashii; lane 4-6, southern blotting using DIG labeled 5.8S rDNA probes. M, marker.

Fig. 2
PCR-RFLP products of the ITS1 gene digested with Rsa I, Alu I, and Msp I enzymes. Lane 1, Metagonimus Miyata type; lane 2, M. yokogawai; lane 3, M. takahashii. M1, M2, marker.

Fig. 3
PCR products for the mCOI gene of three kinds of Metagonimus spp. Lane 1, Metagonimus Miyata type; lane 2, M. yokogawai; lane 3, M. takahashii. M1, M2, marker.

Fig. 4
PCR-RFLP products for the mCOI gene digested with Rsa I, Cfa I, Alu I, and Msp I. Lane 1 & 4, Metagonimus Miyata type; lane 2 & 5, M. yokogawai; lane 3 & 6, M. takahashii. M1, M2, marker.

Fig. 5
Dendrogram of estimated genetic divergence of the three kinds of Metagonimus using program UPGMA v2.0.


Table 1
The sequences of primers used for PCR reaction

Table 2
Proportion of homologous fragments (upper triangle) and estimated genetic divergence (lower triangle)

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