A collection of 38 PVY isolates from seed potato batches, originating from several Western European countries, was characterized by using current biological, serological and molecular tools differentiating PVY strains and groups. The correlation between the three kinds of tests was good but not absolute. No single serological or PCR method was able to discriminate among the five isolate groups found. Twenty-nine isolates belonged to the PVYN strain and six to the PVYO strain. No PVYC was found. Two other isolates reacted serologically like PVYO, but were unable to elicit a hypersensitive response from the Nytbr gene and probably represent the PVYZ group. At the molecular level, these two isolates showed a combination of both PVYO and PVYN and could be recombinants of these strains. Another isolate reacted serologically like PVYO, but induced vein necrosis in tobacco, like PVYN-Wilga. Some PVYN isolates caused tuber ring necrosis in glasshouse conditions. These might belong to the PVYNTN group. The PVYNTN, PVYN-Wilga and PVYZ groups probably represent pathotypes within strains PVYN and PVYO, respectively. The present study also confirms previous reports showing a high genetic variation at the 5′ end within the PVYN strain.