title: Genetic Polymorphisms and Phenotypic Profiles of Sulfadiazine-Resistant and Sensitive Toxoplasma gondii Isolates Obtained from Newborns with Congenital Toxoplasmosis in Minas Gerais, Brazil.
abstract: BACKGROUND
  Previous Toxoplasma gondii studies revealed that mutations in the dhps (dihydropteroate synthase) gene are associated with resistance to sulfonamides. Although Brazilian strains are genotypically different, very limited data are available regarding the susceptibility of strains obtained from human to sulfonamides. The aim of this study was to evaluate the efficacy of sulfadiazine (SDZ) against Brazilian isolates of T. gondii and verify whether isolates present polymorphisms in the dhps gene. We also investigated whether the virulence-phenotype and/or genotype were associated with the profile of susceptibility to SDZ.
  
  
  METHODS
  Five T. gondii isolates obtained from newborns with congenital toxoplasmosis were used to verify susceptibility. Mice were infected with 104 tachyzoites and orally treated with different doses of SDZ. The mortality curve was evaluated by the Log-rank test. The presence of polymorphisms in the dhps gene was verified using sequencing. A descriptive analysis for 11 Brazilian isolates was used to assess the association between susceptibility, genotype, and virulence-phenotype.
  
  
  RESULTS
  Statistical analysis showed that TgCTBr03, 07, 08, and 16 isolates were susceptible to SDZ, whereas TgCTBr11 isolate presented a profile of resistance to SDZ. Nineteen polymorphisms were identified in dhps exons. Seven polymorphisms corresponded to non-synonymous mutations, with four being new mutations, described for the first time in this study. No association was found between the profile of susceptibility and the virulence-phenotype or genotype of the parasite.
  
  
  CONCLUSIONS
  There is a high variability in the susceptibilities of Brazilian T. gondii strains to SDZ, with evidence of drug resistance. Despite the large number of polymorphisms identified, the profile of susceptibility to SDZ was not associated with any of the dhps variants identified in this study. Other genetic factors, not yet determined, may be associated with the resistance to SDZ; thus, further studies are needed as a basis for a more adequate toxoplasmosis treatment.
authors.affiliation: Laboratório de Toxoplasmose, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brasil.
authors.forename: Letícia Azevedo
authors.initials: LA
authors.lastname: Silva
authors.affiliation: Laboratório de Genômica de Parasitos, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brasil.
authors.forename: João Luís
authors.initials: JL
authors.lastname: Reis-Cunha
authors.affiliation: Laboratório de Genômica de Parasitos, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brasil.
authors.forename: Daniella Castanheira
authors.initials: DC
authors.lastname: Bartholomeu
authors.affiliation: Laboratório de Toxoplasmose, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brasil.
authors.forename: Ricardo Wagner Almeida
authors.initials: RW
authors.lastname: Vítor
mesh_terms: D000818:Animals; D000981:Antiprotozoal Agents / Q000494:pharmacology* / Q000627:therapeutic use; D001483:Base Sequence; D001938:Brazil / Q000453:epidemiology; D004094:Dihydropteroate Synthase / Q000235:genetics*; D004351:Drug Resistance / Q000235:genetics; D005260:Female; D056726:Genetic Association Studies; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D051379:Mice; D009154:Mutation; D020641:Polymorphism, Single Nucleotide; D015800:Protozoan Proteins / Q000235:genetics*; D016415:Sequence Alignment; D013411:Sulfadiazine / Q000494:pharmacology* / Q000627:therapeutic use; D014122:Toxoplasma / Q000187:drug effects / Q000235:genetics* / Q000472:pathogenicity; D014124:Toxoplasmosis, Animal / Q000188:drug therapy; D014125:Toxoplasmosis, Congenital / Q000188:drug therapy / Q000453:epidemiology / Q000469:parasitology*; D014774:Virulence / Q000235:genetics
chemical_list: D000981:Antiprotozoal Agents; D015800:Protozoan Proteins; D013411:Sulfadiazine; D004094:Dihydropteroate Synthase