A segment of DNA unique to the kinetoplast of Trypanosoma cruzi was isolated from spontaneously mummified human remains from the coastal area of northern Chile at sites dated from 2000 bc to about ad 1400. Following rehydration of the desiccated human tissue samples of heart, esophagus, or colon, the samples were extracted and primers employed to bind to a 330 bp kinetoplast minicircle DNA sequence present in T. cruzi. This segment was then amplified using the polymerase chain reaction (PCR), and the target segment was visualized by gel electrophoresis. This method enables the identification of Chagas' disease in an ancient body in the absence of recognizable anatomic pathological changes. Am J Phys Anthropol 108:401–407, 1999. © 1999 Wiley-Liss, Inc.