Restriction fragment length polymorphism (RFLP) analysis of total DNA and of ribosomal DNA (rDNA) regions (ribotyping) were used to document Streptococcus pyogenes vertical mother-to-infant transmission and to investigate the spread of S. pyogenes in an obstetric unit. Two isolates from a newborn, two isolates from his mother (patient 1), and two isolates from two other mothers (patients 2 and 3) were studied. RFLP of total DNA, both after HindIII and PvuII digestions and ethidium bromide staining, gave indistinguishable patterns for the strains isolated from the neonate, his mother, and patient 2. Strains from patient 3 and six unrelated strains studied for comparison showed different patterns. In our system, ribotyping was less discriminative than total DNA RFLP analysis. DNA RFLP analysis therefore provides a valuable molecular tool for studying S. pyogenes epidemiology.