Salmonella enterica ssp. enterica serovar Typhimurium and Salmonella enterica ssp. enterica serovar Enteritidis are the major dominating serotypes of Salmonella in poultry and poultry products. Infection by Salmonella Typhimurium is an important cause of morbidity and mortality in poultry. Rapid differentiation of Salmonella Typhimurium from other Salmonella serotypes including Salmonella Enteritidis can be very crutial for public health and for epidemiologists and for the poultry industry. Ten arbitrarily designed short primers (8 to 10 bases) were used in the random amplified polymorphic DNA analysis of Salmonella Typhimurium. One of the primers, primer 3 (5'-CGT GCA CGC-3'), resulted in the amplification of a band pattern that was unique to Salmonella Typhimurium. In total, 24 strains of serotype Salmonella Typhimurium were used during the study. Eighteen of them are clinical isolates, 2 of them chicken isolates (A6, A20), 2 of them from the Pasteur Institute, 1 from Refik Saydam National Culture Collection, and 1 is a type culture strain from National Culture Type Collection. Serotype Salmonella Typhimurium strains, which were collected from several different hospitals, institutes, and culture collections, have all displayed the same amplification band by primer 3. Twenty-three strains of 16 different serotypes of salmoneallae including 11 Salmonella Enteritidis strains gave only a 300-bp amplification band or no bands, whereas an additional 700-bp amplification band was observed only in samples of Salmonella Typhimurium serotype. It is concluded that random amplified polymorphic DNA analysis with primer 3 is of potential use as a serotype-specific marker for Salmonella Typhimurium.