Kinetoplast DNA was isolated from Chilean Trypanosoma cruzi populations and digested with the restriction endonucleases EcoRI, HinfI, HpaII, MspI, and HaeIII. Three major schizodeme groups were discriminated. There was a correlation between the Chilean schizodeme groups (S1, S2, or S3) and the zymodemes known to occur in Chile (Z1, Brazilian Z2 and Bolivian Z2, respectively), although heterogeneity was seen within the schizodeme groups S2 and S3. Standard Brazilian and Bolivian T. cruzi clones (X10 clone 1, Esmeraldo clone 3, SC43 clone 1, and CAN III clone 1) and laboratory strains (Tulahuen and Y) were included in the schizodeme comparisons. SC43 clone 1 had obvious affinities with S3 and X10 clone 1 shared some features with S1 but the other reference stocks could not be definitely assigned to S1, S2, or S3. Fragment patterns and densitometric traces following digestion with HpaII or MspI suggested that kDNA sequences were not methylated.