Aeromonas are gram-negative, oxidase positive, fermenting rods widely distributed in the environment. This genus was recognised as an important fish pathogen, however is often associated with gastrointestinal and extraintestinal diseases in human. Despite its clinical significance Aeromonas spp. are not accurately identified during routine clinical examination, because of their phenotypical similarity. We focused on detailed characterisation of 22 clinical isolates of “A. sobria complex” from diarrhoea samples. Biochemical tests were used to classify isolates into “A. sobria complex”. Further, an approach using ribotyping, whole-cell protein analysis and cpn60 gene sequencing were applied for detailed characterisation and identification. Additionally, 11 reference type strains of Aeromonas from Czech Collection of Microorganisms were included in the comparison. Genetic analysis based on ribotyping clearly distinguished the 22 human isolates from reference Aeromonas type strains and clustered them together with A. veronii bv. sobria, A. veronii bv. veronii and A. allosaccharophila reference strains. A. jandaei and A. schubertii were the next closest relation species. Six isolates were clustered into a separate subclaster(similarity 52%), but results of cpn60 gene sequencing of a selected isolate confirmed A. veronii bv. sobria identification. Whole-cell protein analysis showed different distribution of isolates. 16 isolates were grouped together with A. veronii bv. sobria reference strain (similarity level 48 %) and 8 isolates together with A. veronii bv. veronii reference strain. Other isolates were separated from these groups. Despite inconsistent results of ribotying and whole-cell protein analysis our results showed that studied isolates belonged to “A. sobria complex”, mainly into A. veronii bv. sobria species. Reliable identification of clinical isolates is important for determination of sources of infection.