Fil: Velásquez, Jorge Néstor. Hospital Municipal de Infecciosas Dr. Francisco Javier Muñiz; Argentina.Fil: Pantano, María Laura. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Parasitología; Argentina.Fil: Vittar, Natalia. Hospital Municipal de Infecciosas Dr. Francisco Javier Muñiz; Argentina.Fil: Nigro, Mónica Gabriela. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Parasitología; Argentina.Fil: Figueiras, Olga. Hospital Municipal de Infecciosas Dr. Francisco Javier Muñiz; Argentina.Fil: Astudillo, Osvaldo Germán. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Parasitología; Argentina.Fil: Ricart, Javier. Hospital Municipal de Infecciosas Dr. Francisco Javier Muñiz; Argentina.Fil: Della Paolera, Daniela. Hospital Municipal de Infecciosas Dr. Francisco Javier Muñiz; Argentina.Fil: Carnevale, Silvana. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Parasitología; Argentina.Human cryptosporidiosis is an intestinal infection caused by different species belonging to the genus Cryptosporidium in both immunocompetent and immunocompromised individuals. The life cycle of Cryptosporidium sp. when affecting the digestive system is well known but the infection of other organs is less studied. Molecular methods are necessary for species and subtypes identification. The goal of this work is to propose a new approach that contributes to the diagnosis of the extra-intestinal dissemination process of Cryptosporidium infection. Cryptosporidium sp. was detected in stool and biopsy samples of two HIV-infected patients. DNA was extracted from feces, biopsy specimens, blood, and cerebrospinal fluid (CSF). All samples were analyzed by nested PCR-RFLP of the 18S rDNA, real-time PCR, and gp60 subtyping. Cryptosporidium DNA was detected in stool and tissue samples and it was also present in blood and CSF samples. Both cases were characterized as Cryptosporidium hominis subtype IeA11G3T3. This is the first report that demonstrates the presence of Cryptosporidium DNA in blood and CSF of HIV-infected patients.