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Open Access Methodology article

Updated 16S rRNA-RFLP method for the identification of all currently characterised Arcobacter spp

María José Figueras1*, Arturo Levican1 and Luis Collado2

Author Affiliations

1 Unitat de Microbiologia, Departament de Ciències Mediques Bàsiques, Facultat de Medicina i Ciències de la Salut. IISPV, Universitat Rovira i Virgili, Reus, Spain

2 Institute of Biochemistry and Microbiology, Faculty of Sciences, Universidad Austral de Chile, Valdivia, Chile

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BMC Microbiology 2012, 12:292  doi:10.1186/1471-2180-12-292

Published: 18 December 2012

Abstract

Background

Arcobacter spp. (family Campylobacteraceae) are ubiquitous zoonotic bacteria that are being increasingly recognised as a threat to human health. A previously published 16S rRNA-RFLP Arcobacter spp. identification method produced specific RFLP patterns for the six species described at that time, using a single endonuclease (MseI). The number of characterised Arcobacter species has since risen to 17. The aim of the present study was to update the 16S rRNA-RFLP identification method to include all currently characterised species of Arcobacter.

Results

Digestion of the 16S rRNA gene with the endonuclease MseI produced clear, distinctive patterns for 10 of the 17 species, while the remaining species shared a common or very similar RFLP pattern. Subsequent digestion of the 16S rRNA gene from these species with the endonucleases MnlI and/or BfaI generated species-specific RFLP patterns.

Conclusions

16S rRNA-RFLP analysis identified 17 Arcobacter spp. using either polyacrylamide or agarose gel electrophoresis. Microheterogeneities within the 16S rRNA gene, which interfered with the RFLP identification, were also documented for the first time in this genus, particularly in strains of Arcobacter cryaerophilus isolated from animal faeces and aborted foetuses.

Keywords:
Arcobacter; Identification; Agarose; Polyacrylamide; 16S rRNA-RFLP; 16S rRNA gene mutations