Toxoplasmosis congénita: Diagnóstico serológico, RPC, aislamiento y caracterización molecular de Toxoplasma gondii / Congenital toxoplasmosis: Serology, PCR, parasite isolation and molecular characterization of Toxoplasma gondii

Resumen Introducción El diagnóstico de toxoplasmosis congénita (TC) en el recién nacido es muy importante porque debe recibir tratamiento siempre, sintomático o no, para evitar o aminorar las secuelas de la enfermedad. Objetivo Evaluación comparativa de los métodos disponibles en la institución para el diagnóstico de TC. Materiales y Métodos Se evaluaron métodos diagnósticos en 67 niños cuyas madres cursaron toxoplasmosis aguda durante el embarazo. Se utilizó la técnica de Sabin Feldman para IgG al nacimiento y durante el seguimiento serológico hasta el año de vida. Para determinar IgM, IgA e IgE se utilizó la técnica immunosorbent agglutination assay (ISAGA). El diagnóstico directo se realizó por reacción de polimerasa en cadena (RPC), aislamiento y caracterización molecular del parásito. Resultados La sensibilidad (S) de ISAGA IgM fue 87%, ISAGA IgA 91% y la especificidad (E) fue 100% para ambas; cuando se realizaron en conjunto, la S aumentó a 98%. La detección de IgE contribuyó al diagnóstico cuando se la detectó sólo en la sangre del neonato y no en sangre materna. Se aisló el parásito en cuatro casos de TC, uno fue genotipo II y los otros tres, genotipos "atípicos". La S del aislamiento fue 80% y la E 100%. Conclusión Los métodos serológicos utilizados mostraron una buena eficacia diagnóstica. Un caso fue detectado sólo por el aislamiento y la caracterización molecular tiene gran valor epidemiológico.

Background. Congenital toxoplasmosis diagnosis in the newborn is a very important issue due to the need for early treatment to prevent future sequels. Aim To compare available methods at the institution for the diagnosis of congenital toxoplasmosis. Material and Methods In this study we have evaluated the different diagnostic tests used in 67 congenital exposed newborns, including serological tests, PCR, parasite isolation and molecular characterization. Results The ISAGA IgM and IgA tests showed sensitivity (Se) of 87 and 91%, respectively, and specificity (Sp) of 100%. When ISAGA IgM and IgA were performed simultaneously, the Se increased to 98% and the Sp was 100%. The presence of IgE contributed to the diagnosis when it was detected in the child's serum but not in maternal blood. In four congenital infected children the parasite was isolated and genotyped: one was genotype II and the other three were "atypical" genotypes. No parasite was isolated in children without congenital toxoplasmosis. Discussion Overall, serological tests showed a good diagnostic performance although in one case they were all negative and isolation was the only tool to identify the infection. We conclude that it is essential to use all diagnostic tests in every single exposed child, including if possible, molecular characterization due to its epidemiological implication.

Modelo para la dinámica de transmisión de la toxoplasmosis congénita / A model of congenital toxoplasmosis transmission dynamics

La toxoplasmosis es una zoonosis parasitaria de amplia distribución mundial, que infecta una gran proporción de poblaciones humanas y animales, producida por el parásito Toxoplasma gondii, siendo de gran importancia el contagio de mujeres gestantes, puesto que ésta zoonosis causa enfermedades en los fetos a través de infección transplacentaria; tradicionalmente se ha utilizado una tasa de infección madre-feto constante; sin embargo, hay evidencias de una fuerte relación con la semana de gestación de la madre en el momento de contagiar al feto. Este trabajo describe la dinámica de transmisión de la toxoplasmosis congénita a través de un modelo estructurado en la edad que tiene en cuenta la semana de gestación de la madre; el modelo está acoplado con un modelo espacio-temporal que describe la dispersión de T. gondii a través de gatos, el cual proporciona la cantidad de parásitos en el ambiente, de la cual depende que una mujer gestante se infecte. Se hacen simulaciones variando la cantidad de parásitos y la distancia a la que se encuentra la gestante del lugar del inóculo; además, se varían algunos parámetros equivalentes a posibles medidas de control. Se encuentran cambios importantes al comparar el comportamiento de las poblaciones de gestantes infectadas, cuando se usa tasa de infección madrefeto constante y cuando se usa tasa dependiente de la semana de gestación, lo que permite concluir que la semana de gestación en la que se encuentra la población de gestantes infectadas es fundamental en la transmisión de la infección al feto.

Toxoplasmosis is a parasitic zoonosis having worldwide distribution; it infects many human and animal populations and is produced by the parasite Toxoplasma gondii, this being of great importance in contagion of pregnant women since this zoonosis causes illness in the fetus through transplacental infection. A constant mother-foetus infection rate has traditionally been used. Nevertheless, there is evidence of a strong relationship between a mother's gestation week and the moment when a foetus becomes infected. This work describes congenital toxoplasmosis transmission dynamics by using an age-structured model taking a mother's gestational week into account. The model was adapted to a space-time model describing T. gondii dispersion through cats; this provided the amount of parasites in the environment depending on which a pregnant woman would become infected. Simulations were done, varying the amount of parasites and the pregnant mother-inoculum distance. Some parameters related to possible control measures were also varied. Important changes were found when comparing infected pregnant female population patterns, when a constant mother-fetus infection rate was used and when a dependent pregnancy week rate was used. This led to concluding that the gestation week in which the mothers became infected is fundamental in infection being transmitted to their fetuses.

Significant performance variation among PCR systems in diagnosing congenital toxoplasmosis in São Paulo, Brazil: analysis of 467 amniotic fluid samples

INTRODUCTION: Performance variation among PCR systems in detecting Toxoplasma gondii has been extensively reported and associated with target genes, primer composition, amplification parameters, treatment during pregnancy, host genetic susceptibility and genotypes of different parasites according to geographical characteristics. PATIENTS: A total of 467 amniotic fluid samples from T. gondii IgM- and IgG-positive Brazilian pregnant women being treated for 1 to 6 weeks at the time of amniocentesis (gestational ages of 14 to 25 weeks). METHODS: One nested-B1-PCR and three one-round amplification systems targeted to rDNA, AF146527 and the B1 gene were employed. RESULTS: Of the 467 samples, 189 (40.47 percent) were positive for one-round amplifications: 120 (63.49 percent) for the B1 gene, 24 (12.69 percent) for AF146527, 45 (23.80 percent) for both AF146527 and the B1 gene, and none for rDNA. Fifty previously negative one-round PCR samples were chosen by computer-assisted randomization analysis and re-tested (nested-B1-PCR), during which nine additional cases were detected (9/50 or 18 percent). DISCUSSION: The B1 gene PCR was far more sensitive than the AF146527 PCR, and the rDNA PCR was the least effective even though the rDNA had the most repetitive sequence. Considering that the four amplification systems were equally affected by treatment, that the amplification conditions were optimized for the target genes and that most of the primers have already been reported, it is plausible that the striking differences found among PCR performances could be associated with genetic diversity in patients and/or with different Toxoplasma gondii genotypes occurring in Brazil. CONCLUSION: The use of PCR for the diagnosis of fetal Toxoplasma infections in Brazil should be targeted to the B1 gene when only one gene can be amplified, preferably by nested amplification with primers B22/B23.