DIAGNÓSTICO SOROLÓGICO, ISOLAMENTO E ...

GENERAL ABSTRACT
The overall aim of this study was to compare diverse diagnostic methodologies to toxoplasmosis in
pregnant women from the city of Curitiba. This study was developed in three parts. In the first part,
immunoenzymatic (ELISA) tests were carried out to determine anti-Toxoplasma gondii antibodies
(IgM, IgG) seroprevalence in pregnant women participating at the “Mãe Curitibana” program. In the
second part, isolation procedures to Toxoplasma gondii (in vivo and in vitro) and also
histopathological analysis were done in biological samples (amniotic fluid, blood and placenta) from
pregnant women undergoing recent infection immunological profile. In the third part, other
techniques were evaluated like the polymerase chain reaction (PCR) as a diagnostic method and
the restriction fragment length polymorphisms (RFLP) to determine the genotype(s) of the isolated
parasite(s). During the serological study of 16,479 patients, IgG and IgM were simultaneously
evaluated, and 8,740 patients presented serological results as IgG+ e IgM-; 7203 were IgG- e IgM-;
24 were IgG- e IgM+ and 512 patients were seroreactive to both immunoglobulines. Thirteen
pregnant women showing IgM seroreactivity were submitted to amniocentesis to isolate the
protozoarian. Parasites obtained from the amniotic fluid and blood were isolated in vivo
(intraperitoneal inoculation in mice) and in vitro (cell culture). The optimized PCR protocol used in
this study was efficient to demonstrate the presence of Toxoplasma gondii DNA in all 13 examined
amniotic fluid samples. The molecular characterization by RFLP, indicated that all isolated strains
belonged to genotype-I, confirming data from literature. Diagnostic technical implementations
adopted to gestational toxoplasmosis monitoring will be discussed through this present research
work.
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