title: Genotyping of Toxoplasma gondii: DNA extraction from formalin-fixed paraffin-embedded autopsy tissues from AIDS patients who died by severe disseminated toxoplasmosis.
abstract: This study investigated the genetic features of Toxoplasma gondii isolated directly in autopsies of HIV-infected patients who died with severe disseminated toxoplasmosis. This retrospective analysis was conducted in a cohort of 15 HIV-infected patients with clinical and laboratory data. They had previous cerebral toxoplasmosis at least 6 months before the disseminated toxoplasmosis episode. The hypothesis was that they were infected with highly virulent parasites due to the condition in which they died. T. gondii genotyping was done directly in DNA extracted from 30 autopsy brain and lung samples (2 per patient) and mutilocus PCR-RFLP genotyping was done using 12 molecular markers. The 30 clinical samples were genotyped successfully in 8 or more loci and six suggestive genotypes were identified. One of them was Toxo DB #11, previously identified in different domestic animals and virulent in experimental animals. The other five suggestive genotypes identified in 14 patients were not described. TgHuDis1 was the most frequent and was determined in 8 patients. TgHuDis3 and TgHuDis5 were identified in two patients each. TgHuDis2 and TgHuDis4 have been identified in one patient each. These suggestive genotypes could be considered as virulent, since they caused severe tissue damage and had similar characteristics as Toxo # DB 11.
authors.affiliation: Laboratório de Biologia Molecular de Parasitas e Fungos, Centro de Parasitologia e Micologia, São Paulo, Brazil.
authors.forename: Inara
authors.initials: I
authors.lastname: Bastos da Silva
authors.affiliation: Instituto de Infectologia Emilio Ribas, São Paulo, Brazil.
authors.forename: Tatiana Pimental de Andrade
authors.initials: TP
authors.lastname: Batista
authors.affiliation: Centro de Patologia, Instituto Adolfo Lutz, São Paulo, Brazil.
authors.forename: Roosecelis Brasil
authors.initials: RB
authors.lastname: Martines
authors.affiliation: Centro de Patologia, Instituto Adolfo Lutz, São Paulo, Brazil.
authors.forename: Cristina Takami
authors.initials: CT
authors.lastname: Kanamura
authors.affiliation: Laboratório de Biologia Molecular de Parasitas e Fungos, Centro de Parasitologia e Micologia, São Paulo, Brazil; Supervisão de Vigilância em Saúde de Santo Amaro/Cidade Ademar, Prefeitura Municipal de São Paulo, Brazil.
authors.forename: Isabelle Martins Ribeiro
authors.initials: IM
authors.lastname: Ferreira
authors.affiliation: Instituto de Infectologia Emilio Ribas, São Paulo, Brazil; Departamento de Moléstias Infecciosas e Parasitárias do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo, Brazil.
authors.forename: Jose Ernesto
authors.initials: JE
authors.lastname: Vidal
authors.affiliation: Laboratório de Biologia Molecular de Parasitas e Fungos, Centro de Parasitologia e Micologia, São Paulo, Brazil. Electronic address: pchioccola@gmail.com.
authors.forename: Vera Lucia
authors.initials: VL
authors.lastname: Pereira-Chioccola
mesh_terms: D000163:Acquired Immunodeficiency Syndrome / Q000150:complications* / Q000401:mortality / Q000473:pathology; D000328:Adult; D001344:Autopsy; D001921:Brain / Q000469:parasitology / Q000473:pathology; D001938:Brazil; D015331:Cohort Studies; D016054:DNA, Protozoan / Q000737:chemistry / Q000302:isolation & purification; D005260:Female; D005404:Fixatives; D005557:Formaldehyde; D060005:Genotyping Techniques; D006801:Humans; D007150:Immunohistochemistry; D008168:Lung / Q000469:parasitology / Q000473:pathology; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012189:Retrospective Studies; D014122:Toxoplasma / Q000145:classification* / Q000235:genetics; D014123:Toxoplasmosis / Q000401:mortality* / Q000469:parasitology / Q000473:pathology; D055815:Young Adult
chemical_list: D016054:DNA, Protozoan; D005404:Fixatives; D005557:Formaldehyde
keywords: Brazilian AIDS patients; Disseminated toxoplasmosis; Toxoplasma gondii genotyping