A PCR-RFLP assay for identification of Anisakis simplex from different geographical regions

Anisakis simplex, a nematode from the family Anisakidae, is a parasite of f ish and mammals. It is a casual agent of a human disease called anisakiosis . We found that the assay based on PCR amplification of the ITS-1-5.8 S-ITS -2 fragment of rDNA and subsequent restriction fragment length polymorphism , previously described on the basis of A. simplex isolated solely from one geographical region, can be used as a general test for identification of th is worm species. The restriction patterns analysed for four restriction enz ymes were found to be identical in the case of all A. simplex individuals i solated from as different geographical regions as Baltic Sea, Norwegian Sea , Bering Sea and Sea of Okchotsk. Moreover, our results support the previou sly proposed hypothesis, based on the studies of isoenzymes, that there is a remarkable genetic homogeneity within A. simplex from different geographi cal regions. (C) 2000 Academic Press.