Differentiation of Vibrio vulnificus strains by an arbitrarily primed polymerase chain reaction.

Abstract

A synthetic 17 mer oligonucleotide (5'-GTTGGGTAACGCCAGGG-3') was used as a primer for the arbitrarily primed polymerase chain reaction (AP-PCR) to differentiate various strains of Vibrio vulnificus. A total of 37 genomic DNAs that were extracted from the clinical and environmental strains were successfully differentiated. Among them, 32 profiles of the 37 strains were characterized. None of the environmental and clinical strains had the same amplification profile, suggesting the highly heterogeneous population existed in the strains of V. vulnificus. The size of the amplified sequences ranged from 0.3 to 2.0 Kb and the DNAs were separated to 12 to 20 bands by the 1.2% agarose gel. The clinical isolates from two independent episodes of V. vulnificus infections in a patient were shown to have the same profile, indicating that the second episode was due to recurrence rather than reinfection. The profiles of amplification were reproducible with different preparations of genomic DNA. Arbitrarily primed polymerase chain reaction can therefore be a useful tool for epidemiological study of V. vulnificus infection.