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Genetic and Phenotypic Analysis of Borrelia valaisiana sp. nov. (Borrelia Genomic Groups VS116 and M19)

¹Department of Medical Microbiology, Academic Medical Center, University of Amsterdam, 1105 AZ Amsterdam
⁴Department of Pure and Applied Ecology, University of Amsterdam, 1105 AZ Amsterdam
²The Netherlands; Unité de Bactériologié Moléculare et Médicate, Institut Pasteur, 75724 Paris Cedex 15, France
³Clinical Microbiology, Institut Central des Hôpitaux Valaisans, 1950 Sion, Switzerland

^* Corresponding author. Mailing address: Department of Medical Microbiology, Academic Medical Center, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands, Phone: 31 20 566 4863. Fax: 31 20 697 9271. E-mail: A.P.vanDam{at}amc.uva.nl.

ABSTRACT

To clarify the taxonomic status of two recently described Borrelia genomic groups, groups VS116 and M19, three group VS116 strains and eight group M19 strains isolated from Ixodes ricinus ticks in Switzerland, The Netherlands, and the United Kingdom were characterized. PCR-restriction fragment length polymorphism (RFLP) analysis of the 5S-23S intergenic spacer amplicon, rRNA gene restriction analysis, 16S rRNA gene sequence analysis, randomly amplified polymorphic DNA (RAPD) fingerprinting, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and immunoblotting with monoclonal antibodies were used for genetic and phenotypic analysis. The PCR-RFLP and RAPD patterns of three group VS116 strains and eight group M19 strains were identical but differed from those of Borrelia burgdorferi sensu stricto, Borrelia garinii, Borrelia afzelii, and Borrelia japonica. DNAs from all group VS116 and M19 strains yielded three fragments (6.9, 3.2, and 1.4 kb) and four fragments (2.1, 1.2, 0.8, and 0.6 kb) after digestion with EcoRV and Hindlll, respectively, hybridizing with an Escherichia coli 16S+23S cDNA probe. The SDS-PAGE protein profiles of group VS116 and M19 strains were heterogeneous. Phylogenetic analysis of the partial 16S rRNA gene sequences showed that group VS116 and M19 spirochetes were members of a Borrelia species distinct from previously characterized members of the genus Borrelia. Based on our present study and data from previous DNA-DNA hybridizations, a new Borrelia species, Borrelia valaisiana sp. nov., in the B. burgdorferi complex, is proposed. Strain VS116 is the type strain of this new species.