PCR em tempo real no diagnóstico da toxoplasmose congênita / Real-time PCR in the diagnosis of congenital toxoplasmosis

O diagnóstico da toxoplasmose congênita apresenta limitações sendo, portanto, necessárias novas opções de exames. A análise do líquido aminiótico pela PCR em tempo real já se mostrou eficaz para confirmação da infecção fetal. No entanto, o seu desempenho em outras amostras biológicas ainda não está claro. O objetivo deste estudo é avaliar a PCR em tempo real no sangue da mãe e do recém-nascido assim como no líquido amniótico e placenta, no diagnóstico da toxoplasmose congênita. Esse é um estudo descritivo de gestantes com toxoplasmose acompanhadas no Rio de Janeiro, Brasil. Foi realizada PCR em tempo real em amostras de sangue materno, líquido amniótico, placenta e sangue dos recém-nascidos e o exame histopatológico das placentas. Também foram coletados dados clínicos e laboratoriais dos recém-nascidos. Foram acompanhadas 116 gestantes e analisadas 298 amostras. Uma (0,9%) gestante apresentou PCR positiva no sangue, três (3,5%) no líquido amniótico, uma (2,3%) na placenta e nenhum recém-nascido apresentou PCR positiva no sangue. O estudo histopatológico foi sugestivo de infecção por toxoplasmose em 24 (49%) placentas. Seis (5,2%) recém-nascidos foram diagnosticados com toxoplasmose congênita e apenas os casos com PCR positiva no líquido amniótico tinham associação do resultado da PCR com o diagnóstico de infecção congênita. Tanto as amostras de sangue materno quanto as de sangue dos recém-nascidos e placenta, não demonstraram ser promissoras no diagnóstico da toxoplasmose congênita. Novos estudos são necessários para avaliar o real papel do diagnóstico molecular em outros materiais biológicos que não o líquido amniótico.

The diagnosis of congenital toxoplasmosis has limitations so new options are needed. Real-time PCR analysis of amniotic fluid has proven effective for confirming fetal infection. However, its performance in other biological samples still needs to be determined. This study aims to evaluate the real-time PCR role in the blood of the mother and newborn as well as in the amniotic fluid and placenta, in congenital toxoplasmosis diagnosis. It is a descriptive study of pregnant women with toxoplasmosis followed in Rio de Janeiro, Brazil. Real-time PCR was performed on maternal blood, amniotic fluid, placenta, and newborn blood samples. In addition, a histopathological examination of the placentas was performed and data from the babies were collected. One hundred and sixteen pregnant women were followed and 298 samples were analyzed. One (0.9%) pregnant woman had positive PCR in the blood, three (3.5%) in the amniotic fluid, one (2.3%) in the placenta, and any newborn had positive PCR in the blood. The histopathological study suggested toxoplasmosis infection in 24 (49%) placentas. Six (5.2%) newborns were diagnosed with congenital toxoplasmosis and only the cases with positive PCR in amniotic fluid associated with the diagnosis of congenital infection. Neither maternal nor newborn blood and placenta samples have not shown promise in diagnosing congenital toxoplasmosis. Further studies are needed to evaluate the fundamental role of molecular diagnostics in others biological materials than amniotic fluid.

Epidemiological profile of 39 cases of microcephaly caused by congenital infections diagnosed in the state of Rio Grande do Sul between 2015-2017

Introduction: Microcephaly is a clinical finding that can arise from congenital anomalies or emerge after childbirth. Maternal infections acquired during pregnancy can result in characteristic brain damage in the newborn (NB), which may be visible even in the fetal stage. To describe the epidemiological profile of newborns with reported microcephaly and diagnosed with congenital infections in the state of Rio Grande do Sul between 2015 and 2017. Methods: A cross-sectional study was carried out on data collected from the Public Health Event Registry as well as from medical records. The investigation included serologies for toxoplasmosis and rubella; polymerase chain reaction (PCR) for Zika virus (ZIKV) in the blood and cytomegalovirus in the urine; non-treponemal tests for syphilis; and brain imaging tests. Results: Of the 257 reported cases of microcephaly, 39 were diagnosed with congenital infections. Severe microcephaly was identified in 13 patients (33.3%) and 51.3% of the cases showed alterations in brain imaging tests. In relation to the diagnosis of congenital infections, three patients (7.7%) were diagnosed with ZIKV, nine (23.1%) with cytomegalovirus, nine (23.1%) with toxoplasmosis, and 18 (46.1%) with congenital syphilis. The three cases of ZIKV showed calcification in brain imaging tests, signs of arthrogryposis, excess occipital skin and irritability, characterizing the typical phenotype of ZIKV infection. Conclusions: Most cases of congenital infection had severe neurological lesions, particularly the cases of ZIKV, which can cause neurodevelopmental delays and sequelae in these infants throughout early childhood.

Prevalence of congenital toxoplasmosis among a series of Turkish women / Prevalencia de toxoplasmosis congénita en una serie de mujeres en Turquía

Background: Toxoplasma gondii infection during pregnancy causes congenital malformations. Pregnant women should be screened for this infection since it is preventable and treatable. Aim: To study the sero prevalence of Toxoplasma gondii infection among pregnant women living in lzmir, Turkey. Material and Methods: A blood sample was obtained from 4651 women aged between 15 and 45years, during their first trimester of pregnancy. IgM and IgG antibodies against Toxoplasma gondii were measured using an ELISA assay. Among women with both IgG and IgM antibodies positive, an IgG avidity test was performed, using a VIDAS kit. Results: IgG antibodies were positive in 1871 (39.9%) participants. Of these, 48 (2.5%) also had positive IgM antibodies. In 41 ofthese 48 women, the IgG avidity test was performed and only one woman had a low avidity. This woman was treated with Spiramycin. Her offspring had an intrauterine growth retardation and oligohydramnios. A chorioretinitis was diagnosed in the offspring of other woman with both antibodies positive. Conclusions: In this series, the prevalence of congenital toxoplasmosis was low. However, women with positive antibodies against Toxoplasma Gondii should be further studied and followed during their pregnancy.

Antecedentes: La infección por Toxoplasma gondii durante el embarazo causa malformaciones congénitas. Se debe efectuar serologíapara esta infección en mujeres embarazadas ya que es prevenible y tratable. Objetivo: Estudiar la seroprevalencia de infección por Toxoplasma gondii en mujeres embarazadas que viven en Esmirna, Turquía. Material y Métodos: Se obtuvo una muestra de sangre en 4.651 mujeres cuyas edades fluctuaban entre 15 y 45, años, durante su primer trimestre de embarazo. Los anticuerpos IgM e IgG en contra de Toxoplasma gondii se midieron por ELISA. En mujeres que tenían anticuerpos IgG e IgM positivos, un ensayo de avidez de IgG se efectuó utilizando el kit VIDAS. Resultados: Los anticuerpos IgG fueron positivos en 1.871 participantes (39,9%). De estas, 48 (2,5%) también tenían anticuerpos IgM positivos. En 41 de estas 48 mujeres, se efectuó el test de avidez y sólo una tenía una baja avidez. Esta mujer se trató con espiramicina y su producto de concepción tuvo un retardo de crecimiento intrauterino y un oligohidroamnios. Una corioretinitis se diagnosticó en el producto de concepción de otra mujer con ambos anticuerpos positivos. Conclusiones: La seroprevalencia de toxoplasmosis congénita en esta serie de pacientes fue baja, sin embargo, las mujeres con anticuerpos positivos deben ser tratadas y seguidas.

Perfil sorológico para toxoplasmose em gestantes atendidas em maternidade / Serologic profile of toxoplasmosis in pregnant women attended at a teaching-hospital in Recife

OBJETIVOS: Determinar o perfil sorológico para toxoplasmose e identificar os principais fatores associados à susceptibilidade (pacientes com imunoglobulinas IgG e IgM ausentes) em gestantes atendidas em uma maternidade-escola do Recife. MÉTODOS: Realizou-se um estudo de corte transversal, incluindo 503 gestantes submetidas à sorologia para toxoplasmose no IMIP (Recife), no período de outubro de 2004 a abril de 2005. Realizou-se imunofluorescência indireta para pesquisa de IgG e IgM e um breve questionário foi aplicado às pacientes, descrevendo-se identificação, características demográficas e obstétricas, antecedentes mórbidos relevantes, hábitos de vida e tipo de moradia. Para análise estatística, utilizaram-se os testes Qui quadrado de associação e exato de Fisher, com um nível de significância de 5 por cento. RESULTADOS: Constatou-se imunidade para toxoplasmose em 74,7 por cento, susceptibilidade em 22,5 por cento e "possível" infecção ativa em 2,8 por cento das gestantes. Não se encontrou associação estatisticamente significativa entre susceptibilidade para toxoplasmose e idade, procedência, condições mórbidas, hábitos, condições de habitação, rede de esgotos, criação de animais domésticos, número de gestações e idade gestacional. Verificou-se uma associação significativa entre susceptibilidade para toxoplasmose e escolaridade, com uma maior freqüência de susceptibilidade entre mulheres com oito ou mais anos de estudo. CONCLUSÃO: A freqüência de susceptibilidade para toxoplasmose é relativamente baixa entre pacientes atendidas no pré-natal em nosso meio e nenhum outro fator preditivo além da escolaridade foi identificado.

OBJECTIVES: To determine the serologic profile of toxoplasmosis and the main factors associated with susceptibility (patients without IgM and IgG antibodies) in pregnant women attended at a teaching-hospital in Recife, Brasil. METHODS: A cross-sectional study was carried out, enrolling 503 pregnant women submitted to serology for toxoplasmosis at IMIP (Recife) from October 2004 to April 2005. Anti-Toxoplasma IgG and IgM antibodies were studied by IFA. A short questionnaire was administered to patients to provide identification, demographic and obstetrical characteristics, past history of morbidity, habits and dwelling conditions. The chi-square and Fisher-exact tests were used at a 5 percent level of significance. RESULTS: Immunity for toxoplasmosis was present in 74.7 percent, susceptibility in 22.5 percent and "possible" active infection in 2.8 percent of patients. No significant associations were observed between toxoplasmosis susceptibility and age, location, conditions of morbidity, habits, dwelling conditions and sewage system, living with animals, pregnancy and gestational age. A significant association between toxoplasmosis susceptibility and schooling was found, with a higher frequency of susceptibility among women with eight or more years of schooling. CONCLUSION: Susceptibility for toxoplasmosis was relatively low in these prenatal patients and schooling was the only identifiable predictive factor.

Construções e redefinições: as toxoplasmoses / Constructions and redefinitions: the toxoplasmosis

Esta tese tem como objeto o processo de redefinição da categoria diagnóstica infecção/doença Toxoplasmose. É um estudo qualitativo, inserido no campo science studies, de abordagem crítica reformista e de natureza empírico-analítica. Vincula-se à linha de pesquisa intitulada Instituições, saberes e práticas em saúde e ao projeto Os médicos e a ciência do Instituto de Medicina Social da Universidade do Estado do Rio de Janeiro. As unidades de análise foram: 1) Agentes envolvidos na produção do conhecimento científico em nível local; 2) Documentos normativos locais; 3) Documentos normativos nacionais e internacionais; 4) Instituição: Laboratório Reconhecer do Centro de Biociências e Biotecnologia (CBB), da Universidade Estadual do Norte Fluminense Darcy Ribeiro (UENF/Darcy Ribeiro). Como técnicas de pesquisa foram utilizadas a observação etnográfica, entrevistas e pesquisa documental da produção científica. Os pressupostos foram de que essa redefinição seja decorrente de uma construção (Hacking, 1999; Latour, 1997, 2000, 2001) e realizada em uma arena transepistêmica (Knor-Cetina, 1981). Foram acrescidos a esses conceitos, os de referência circulante (Latour, 2001) e os da taxonomia dos elementos dos objetos da ciência laboratorial, ou seja, os conceitos idéias, marcas e coisas (Hacking,1992). Considerando essa dinâmica, as redefinições em relação a essa infecção/doença estariam em um período de pouca estabilização, embora elas não se definiriam completa e eternamente pela dependência que possuem do invólucro espaço-temporal (Latour, 2001) e da referência circulante.

This thesis has as object the process of redefinition of the disgnostic categoryinfection/illness "Toxoplasmosis". It is a qualitative, inserted study in the field "science studies", of reformist critical boarding and empiricist- analytical nature. The line of intitledresearch is associated to it "Institutions, to know and practical in health" and to the project "the doctors and the science" of the Institute of Social Medicine of the University of the Stateof Rio De Janeiro. The units of analysis had been: 1) "involved Agents" in the production of the scientific knowledge in local level; 2) local normative Documents; 3) national and international normative Documents; 4) Institution: Laboratory To recognize of the Center of Biosciences and Biotechnology (CBB), of the State University of the Of the state of Rio deJaneiro North Darcy Ribeir (UENF/Darcy Ribeiro). As research techniques had been used the etnografic observation, interviews and documentary research of scientific production. The estimated ones had been of that this redefinition is decurrent of a "construction" (Hacking,1999; Latour, 1997, 2000, 2001) and carried through in a "transepistemic arenas" (Knor-Cetina, 1981). They had been increased to these concepts, of “circulating reference” (Latour, 2001) and of the taxonomy of the elements of objects of laboratorial science, that is, the concepts "ideas", “marks” and “things” (Hacking, 1992). Considering this dynamics, the redefinitions in relation to this infection/illness would be in a period of little stabilization, even so they are not defined completely and perpetual for the dependence that possess of the "pack space-weather" (Latour, 2001) and of the "circulating reference".

Congenital toxoplasmosis in two health institutions in Trinidad / Toxoplasmosis congénita en dos centros de salud de Trinidad

Toxoplasmosis is the most widespread zoonosis and an important human disease particularly in children where it could cause visual and neurological impairment and mental retardation. This study was conducted to determine the prevalence of toxoplasmosis, especially congenital toxoplasmosis in patients at two health institutions in Trinidad A total of 504 cord blood samples of newborn babies were collected: 174 from a women's hospital and 330 from a general hospital. In order to elicit aternal and prenatal risk factors for toxoplasmosis, mothers of the newborns completed a questionnaire. Enzyme-immuno assay (EIA) was used to detect IgG and IgM to Toxoplasma gondii. Overall, of 504 serum samples tested, 220 (43.7%) were seropositive for IgG while the prevalence of congenital toxoplasmosis as reflected by IgM was 0.4%. The prevalence of IgG and IgM by health institutions was not significantly different (p > 0.05; chi-square). The prevalence of toxoplasmosis using IgG was highest in neonates of mothers who were of East Indian descent (54.1%), had four children (52.9%), kept cats in households (47.7%), practised outdoor gardening (50.8%), consumed raw meat (66.7%), had experienced miscarriage(s) (47.3%), stillbirths (66.7%), or who had eye problem(s) (52.9%) and mental retardation (50.0%). The study prevalence of congenital toxoplasmosis revealed a high seroprevalence oftoxoplasmosis in neonates but there was 0.4% serological evidence of congenital disease. It indicates a need for sensitization of the population and healthcare workers and for follow-up of infected children for clinical evidence of the disease. This would be necessary to fully appreciate the impact of toxoplasmosis in Trinidad and Tobago. The differences from comparison groups were however not statistically significant (p > 0.05; chi-square).

La toxoplasmosis es la zoonosis más extendida y una enfermedad humana importante, particularmente en niños, a quienes puede causar daño visual y neurológico, y retraso mental. Este estudio se llevó a cabo con el propósito de determinar la prevalencia de la toxoplasmosis, especialmente la toxoplasmosis congénita en pacientes de dos centros de salud en Trinidad. Se recogieron un total de 504 muestras de sangre de cordón umbilical de neonatos: 174 de mujeres en un hospital de mujeres y 330 en un hospital general. A fin de obtener información sobre los factores de riesgo maternos y prenatales en relación con la toxoplasmosis, las madres de los recién nacidos llenaron una encuesta. Un ensayo inmunoenzimático (EIE) fue usado para detectar anticuerpos IgG e IgM contra el Toxoplasma gondii. En general, de 504 muestras de suero examinadas, 220 (43.7%) resultaron seropositivas al IgG, mientras que la prevalencia de la toxoplasmosis congénita reflejada por el IgM fue 0.4%. La prevalencia de IgG e IgM por parte de las instituciones de salud no fue significativamente diferente (p > 0.05; chi-cuadrado). La prevalencia de la toxoplasmosis usando IgG fue más alta en los neonatos cuyas madres eran ascendencia indoriental (54.1%), tenían cuatro niños (52.9%), mantenían gatos en sus casas (47.7%), practicaban jardinería al aire libre (50.8%), consumían carne cruda (66.7%), habían tenido aborto(s) (47.3%), partos de feto muerto (66.7%), o tenían problema(s) de los ojos (52.9%) y retardo mental (50.0%). Este estudio de la toxoplasmosis congénita, reveló una alta seroprevalencia de toxoplasmosis en neonatos, pero hubo 0.4% de evidencia serológica de enfermedad congénita. Esto apunta a la necesidad de sensibilizar a la población y a los trabajadores del cuidado de la salud, e igualmente indica la necesidad de realizar seguimientos a los niños infectados, en busca de evidencia clínica de la enfermedad. Esto es necesario si se quiera valorar totalmente el impacto de la...

Otimizaçäo da reaçäo de polimerase em cadeia para detecçäo de Toxoplasma gondii em sangue venoso e placenta de gestantes / Optimization of Toxoplasma gondii detection by PCR in pregnants blood and placenta

A detecçäo de Toxoplasma gondii no sangue venoso e na placenta de gestantes pela reaçäo de polimerase em cadeia pode facilitar o diagnóstico préðnatal do toxoplasmose congênita. Foram avaliadas gestantes IgMðreagentes e os seus filhos. Além das dosagens de IgG, IgM, IgA e reaçäo de avidez de IgG (MEIA), foram realizadas a técnica de imunoperoxidase e a inoculaçäo em camundongos. De cada amostra foi efetuada amplificaçäo gênica com primers do gene B1 e novos primers do gene TGR (chamados ABGTg7 C1 e N1). É preciso observar que o tratamento poderia ser responsável por uma diminuiçäo da infecçäo. Desta forma, o diagnóstico negativo confirmaria a eficiência do tratamento preventivo na replicaçäo parasitária no útero. A reaçäo de polimerase em cadeia mostrouðse sensível e específica; evidenciou a presença de um a dez taquizoítas; pode ser utilizada com segurança e confiabilidade, além de tornar rápido o diagnóstico da toxoplasmose congênita, sendo, assim, ferramenta importante na avaliaçäo préðnatal