Genomic sequences in VP1/2A and 5'-non-coding region of 10 isolates of Coxsackievirus B5 from three outbreaks were compared with published sequences of another Coxsackievirus B5, swine vesicular disease virus, Coxsackievirus B1, Coxsackievirus B3, and Coxsackievirus B4. Isolates of Coxsackievirus B5 from the same outbreak showed close relations, not exceeding 7.2% in nucleotide differences. Differences were greater between isolates from different outbreaks, varying between 8.4 and 16%. We have also shown that Coxsackie B5 viruses from an outbreak in 1967 are more similar to viruses from an outbreak in 1983 than to the viruses isolated from an intervening outbreak in 1972. The sequence comparison of Coxsackievirus B5 isolates with other Coxsackie B viruses and swine vesicular study, cDNA synthesis, polymerase chain reaction, and sequencing, are suitable for rapid Coxsackie B virus detection and identification of genotypic relations between viruses originating from different outbreaks.