title: Fitness of Toxoplasma gondii is not related to DHFR single-nucleotide polymorphism during congenital toxoplasmosis.
abstract: Factors that regulate the pathogenesis of Toxoplasma gondii in humans are poorly understood. When acquired during pregnancy, toxoplasmosis can be disastrous, leading to fetal loss or conversely to subclinical disease. In congenitally infected infants, evolution is highly unpredictable. Genotype based virulence patterns have been described in mice, but in humans this classification does not correlate with the gravity of the disease. Mutations on DHFR-TS loci have recently been reported to confer T. gondii fitness cost. In this study, we investigated the relationship between the virulence of the parasite, as measured by clinical outcome in the fetus or newborn, fitness, as measured by parasitic load in amniotic fluid, and allelic polymorphism in DHFR. Six cases of severe congenital toxoplasmosis and 23 cases of mild congenital infections were included in the study. Quantitative PCR was performed to evaluate total T. gondii DNA load in amniotic fluid and detection of mutations was carried out with a LightCycler using hybridisation probes. Parasitic load was significantly higher in severe infections than in mild diseases. Among isolates from severe or non-severe cases of congenital toxoplasmosis, no polymorphism could be detected at loci 36, 83 or 245 of the DHFR gene. The virulent RH strain presented the same melting temperature as the non-virulent PRU strain for codons 36, 83 and 245. Only mutated clones, M2M3 and M2M4 with allelic replacement at these positions, displayed different profiles allowing a clear distinction between wild and mutant types. We concluded that the DHFR gene mutations we investigated do not regulate T. gondii fitness in humans.
authors.affiliation: E.A. 3732, Parasitology Unit, Faculty of Medicine, Claude Bernard University, Lyon, France. francois.peyron@chu-lyon.fr
authors.forename: François
authors.initials: F
authors.lastname: Peyron
authors.forename: Nathalie
authors.initials: N
authors.lastname: Eudes
authors.forename: Frédérique
authors.initials: F
authors.lastname: de Monbrison
authors.forename: Martine
authors.initials: M
authors.lastname: Wallon
authors.forename: Stéphane
authors.initials: S
authors.lastname: Picot
mesh_terms: D000818:Animals; D001483:Base Sequence; D016054:DNA, Protozoan / Q000032:analysis; D004351:Drug Resistance / Q000235:genetics; D017125:Genes, Protozoan; D005819:Genetic Markers; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods; D007231:Infant, Newborn; D008969:Molecular Sequence Data; D020641:Polymorphism, Single Nucleotide; D016415:Sequence Alignment; D014122:Toxoplasma / Q000502:physiology*; D014125:Toxoplasmosis, Congenital / Q000188:drug therapy / Q000469:parasitology*; D014774:Virulence
chemical_list: D016054:DNA, Protozoan; D005819:Genetic Markers