Typification was PCR based on Nicaragua isolate DNA (Nic) amplified with primers flanking several T. cruzi DNA markers. Fig 1A: The intergenic miniexon of the spliced leader genes was amplified with TC, TC1 or TC2 primers showing a 350 bp fragment for Nic and Sylvio X-10 clone (Tc I) DNA. Fig. 1B: The A10 fragment was amplified with P3 and P6 primers, and a 657 bp band was amplified for Tul2 and CL Brener DNA; no DNA band was observed for Nic, Sylvio and Y (Tc II), as was expected for lineages I and II. Fig 1C: The 24 S alpha rDNA marker was amplified with D71, D75 and D76 primers, and a 125 bp product was visualized for Nic and Sylvio.