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One-step polymerase chain reaction-based typing of Arcobacter species
Hidenori Kabeya1, Yoko Kobayashi, Soichi Maruyama
1Laboratory of Veterinary Public Health, Department of Veterinary Medicine, College of Bioresource Sciences, Nihon University, Kameino, Fujisawa, Kanagawa 252-8510, Japan. kabeya@brs.nihon-u.ac.jp
Abstract:
A species-specific PCR assay was developed for the identification of the Arcobacter species, Arcobacter butzleri, Arcobacter cryaerophilus 1A and 1B, and Arcobacter skirrowii. The primers, which amplify the most variable areas of the 23S rRNA gene, were designed to perform species-specific identification by one-step PCR. The DNA sequence of the region from A. cryaerophilus 1B was determined, and the specific pimer for the species was designed. By using one-step PCR containing the mixed primers N.butz, N.c1.A, N.c.1B, and N.ski, species-specific amplifications were detected from the reference strains of A. butzleri, A. cryaerophilus 1A, 1B, and A. skirrowii, respectively. Primers designed in this study were also evaluated on 10 of Japanese field isolates, and all species were identified. This simple one-step PCR assay was found to be a powerful tool for the survey of Arcobacter infection.