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A sensitive and specific PCR hybridization assay was developed for the simultaneous detection and identification ofEhrlichia andBorrelia burgdorferi sensu lato. In separate assays the 16S rRNA gene ofEhrlichia species and the 23S-5S rRNA spacer region ofB. burgdorferi sensu lato were amplified and labeled by PCR. These PCR products were used in a reverse line blot hybridization assay in which oligonucleotide probes are covalently linked to a membrane in parallel lines. Hybridization of the samples with the oligonucleotide probes on this membrane enabled the simultaneous detection and identification ofEhrlichia ,B. burgdorferi , andBartonella species in 40 different samples. The application of the assay to DNA extracts from 121Ixodes ricinus ticks collected from roe deer demonstrated that 45% of these ticks carriedEhrlichia DNA. More than half of these positive ticks carried species with 16S rRNA gene sequences closely related to those ofE. phagocytophila and the human granulocytic ehrlichiosis agent. The majority of the other positive ticks were infected with a newly identifiedEhrlichia -like species. In addition, 13% of the ticks were infected with one or moreB. burgdorferi genospecies. In more than 70% of the ticks 16S rRNA gene sequences forBartonella species or other species closely related toBartonella were found. In five of the ticks bothEhrlichia andB. burgdorferi species were detected.

Detection and Identification ofEhrlichia,Borrelia burgdorferiSensu Lato, andBartonellaSpecies in DutchIxodes ricinusTicks