We examined 310 strains of Listeria monocytogenes by multilocus enzyme electrophoresis. Fifty-six electrophoretic types (ETs) of the organism were defined: 10 for serovar 4b strains, 11 for serovar 1/2b strains, and 30 for serovar 1/2a strains. Strains of serovars 1/2c, 3a, and 3b, and a non-typable strain were distributed among the remaining five ETs. The mean genetic diversity of the species was 0.41. Principal coordinate analysis revealed a sharp division among ETs which divided the species into two major clusters. ETs containing serovar 1/2a strains were in one cluster while all ETs containing serovar 4b, 1/2b, and 3b strains were in the second cluster. Except for two ETs that contained strains from both serovar 1/2b and serovar 3b, no ET contained strains from more than one serovar. Multilocus enzyme electrophoresis facilitated the analysis of epidemiologic data. In three separate epidemiologic investigations electrophoretic typing confirmed a common source as a cause of an outbreak; in a fourth investigation a single common source as a cause of an outbreak was effectively ruled out. Electrophoretic typing was also useful in documenting potential links between Listeria contaminated foods and persons with listeriosis who consumed those foods.