title: Genotyping of Toxoplasma gondii Directly from Human and Animal Biological Samples: From Partial Genotypes to a New Genotype.
abstract: Genotyping of Toxoplasma gondii is traditionally performed using DNA obtained from tachyzoites after isolation by bioassay in mice. In this study, genotyping of T. gondii was performed by multiplex nested polymerase chain reaction restriction fragment length polymorphism (Mn-PCR-RFLP) in DNA obtained from the lungs of experimentally infected mice, the hearts of naturally infected free-range chickens, and human blood samples of newborns with congenital toxoplasmosis. The efficiency of Mn-PCR varied according to the marker. We obtained complete genotypes of all of the mice lung samples. In chickens, total or partial genotyping was performed on all of the 15 samples. Two complete genotypes were obtained, including one identified for the first time, and another previously described in different hosts including dogs, cats, and humans. In blood from infants, partial genotypes were obtained in 8 of the 12 samples. Mouse bioassay is the most efficient method to obtain DNA from T. gondii , but direct tissue genotyping enhances the likelihood of obtaining molecular information on T. gondii and is an effective tool as a complement to isolation in mice. In this study, we genotyped Toxoplasma gondii directly from human (blood samples of newborns with congenital toxoplasmosis) and free-range chickens (hearts) by Mn-PCR-RFLP. We present partial and complete genotypes and provide technical and scientific information about T. gondii genotyping methods.
authors.affiliation: Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, 31270-901, Belo Horizonte, MG, Brazil.
authors.forename: M M
authors.initials: MM
authors.lastname: Cunha
authors.affiliation: Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, 31270-901, Belo Horizonte, MG, Brazil.
authors.forename: A C A V
authors.initials: AC
authors.lastname: Carneiro
authors.affiliation: Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, 31270-901, Belo Horizonte, MG, Brazil.
authors.forename: J G L
authors.initials: JG
authors.lastname: Costa
authors.affiliation: Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, 31270-901, Belo Horizonte, MG, Brazil.
authors.forename: R W A
authors.initials: RW
authors.lastname: Vitor
mesh_terms: D000653:Amniotic Fluid / Q000737:chemistry; D000818:Animals; D001681:Biological Assay; D001938:Brazil; D002415:Cats; D002645:Chickens; D016054:DNA, Protozoan / Q000097:blood / Q000134:cerebrospinal fluid; D004285:Dogs; D005819:Genetic Markers; D005838:Genotype; D060005:Genotyping Techniques / Q000379:methods*; D006801:Humans; D007231:Infant, Newborn; D051379:Mice; D014122:Toxoplasma / Q000145:classification* / Q000235:genetics; D014124:Toxoplasmosis, Animal / Q000469:parasitology*; D014125:Toxoplasmosis, Congenital / Q000469:parasitology*
chemical_list: D016054:DNA, Protozoan; D005819:Genetic Markers