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Skin biopsies taken from UK cases of erythema migrans rash were cultured for Borrelia burgdorferi sensu lato. Reverse line blotting was used to type the infecting genospecies in PCR-positive cultures and biopsies. B. garinii or B. afzelii was identified in 56% (5/9) of biopsies/cultures tested. All patients were tested by conventional serology. PCR confirmed infection in two patients where serological testing failed to detect antibody.