The research comprises isolation and identification of Bifidobacterium strains from faeces of breast-fed infants. Modified Garche’s agar medium, containing selective (penicillin and lithium chloride) or stimulating substances (maltodextrine instead of lactose), and anaerobic culture conditions (Gas Pak H₂ + CO₂, Oxoid) were used for strain isolation. Identification of isolates to the genus was based on fructose-6-phosphate phosphoketolase activity, lack of ability to produce catalase, indol from tryptofan, gas from glucose, and to reduce nitrate. Species identification was performed using PCR technique applying 16S rDNA-gene-targeted primers specific to B. infantis, B. breve and B. longum. Type strains from American Type Culture Collection (ATCC) were used as reference. As a result of the studies, 14 Bifidobacterium strains were isolated, among which 7 were classified to B. breve and 6 to B. longum species. B. infantis was not isolated. It was also impossible to identify Bifidobacterium sp. N10 in the applied system of primers.