MULTIPLEX PCR FOR THE IDENTIFICATION OF ARCOBACTER AND DIFFERENTIATION OF ARCOBACTER-BUTZLERI FROM OTHER ARCOBACTERS

A multiplex polymerase chain reaction (PCR) assay to identify Arcobact er isolates and to distinguish A. butzleri, from other arcobacters is described. The test uses two primer sets. Set I targets a section of t he 16S rRNA genes of Arcobacter spp. Set II amplifies a portion of the 23S rRNA genes unique to A. butzleri. Specificity of the primer sets was evaluated using ATCC reference strains of A. butzleri, A. cryaerop hilus, A, skirrowii, Bacteroides spp., Campylobacter ro spp., Helicoba cter spp. and Wolinella succinogenes. Upon PCR amplification, all of t he Arcobacter isolates yielded a 1233 bp product, whereas A. butzleri ATCC 49616 exhibited both a 1223 bp and a 686 bp product. No PCR produ ct was observed for other closely related ATCC strains (n = 37). We ne xt analyzed by multiplex PCR field strains of Arcobacter spp. (it = 10 8) which had been previously characterized to the species level by eit her DNA-DNA hybridization, dot blot hybridization, ribotyping or by se rology. The 1223 bp multiplex PCR product identified all of the isolat es as Arcobacter. The presence of both the 1223 and 686 bp amplicons i dentified 66 strains as A. butzleri. Speciation by multiplex PCR agree d with results obtained by the other methods. The multiplex PCR assay is specific, rapid and easy to interpret and, thus, will aid in elucid ating the prevalence, epidemiology and zoonotic potential of Arcobacte r. Published by Elsevier Science B.V.