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Objective The purpose of the present study is to standardize and evaluate the use of the immunoglobulin G (IgG) antibody avidity test on blood samples from newborns collected on filter paper to perform the heel test aiming at its implementation in ongoing programs. Methods Blood samples from newborns were collected on filter paper simultaneously with the heel prick test. All samples were subjected to immunoglobulin M IgM and IgG enzyme-linked immunosorbent assays (ELISA). Peripheral blood was collected again in the traditional way and on filter paper from newborns with high IgG levels (33). Three types of techniques were performed, the standard for measuring IgG in serum, adapted for filter paper and the technique of IgG avidity in serum and on filter paper. The results of the avidity test were classified according to the Rahbari protocol. Results Among the 177 samples, 17 were collected in duplicate from the same child, 1 of peripheral blood and 1 on filter paper. In this analysis, 1 (5.88%) of the 17 samples collected in duplicate also exhibited low IgG avidity, suggesting congenital infection. In addition, the results obtained from serum and filter paper were in agreement, that is, 16 (94.12%) samples presented high avidity, with 100% agreement between the results obtained from serum and from filter paper. Conclusion The results of the present study indicate that the avidity test may be another valuable method for the diagnosis of congenital toxoplasmosis in newborns. Resumo Objetivo O objetivo do presente estudo é padronizar e avaliar a utilização do teste de avidez de anticorpos imunoglobulina G (IgG) em amostras de sangue de recémnascidos (RNs) coletadas em papel filtro para a realização do teste do pezinho visando a implementação nos programas já vigentes. Métodos Foram coletadas amostras de sangue de recém-nascidos em papel filtro simultaneamente ao teste do pezinho. Em todas as amostras, foram realizados os testes imunoenzimáticos (ELISA) imunoglobulina M (IgM) e IgG. Dos RNs que apresentaram altos índices de IgG (33), foi novamente coletado sangue periférico da forma tradicional e em papel filtro. Foram realizadas técnicas padrão para a dosagem de IgG em soro, adaptadas para papel filtro, e a técnica de avidez de IgG em soro e em papel filtro. Os valores obtidos para o teste de avidez foram classificados de acordo com o protocolo de Rahbari. Resultados Dentre as 177 recoletas, em 17 amostras foi realizada a coleta simultânea de sangue periférico e papel filtro da mesma criança. Nesta análise, 1 (5,88%) das 17 amostras coletadas em duplicata obteve também baixa avidez de IgG, sugerindo infecção congênita da criança, e houve concordância entre os resultados obtidos em soro e em papel filtro: 16 (94,12%) das amostras apresentaram alta avidez, com concordância de 100% entre os resultados obtidos em soro e em papel filtro. Conclusão Os dados do presente trabalho evidenciam que o teste de avidez poderá ser mais um método valioso a ser utilizado no diagnóstico da toxoplasmose congênita em RNs.

The aim of this study was to evaluate the genotypic characteristics of Toxoplasma gondii isolated from free-range chickens in the metropolitan area of Goiânia, Goiás, in Brazil’s central-west region. The seroprevalence rate was found to be 96%, according to an indirect hemagglutination assay. Brain and heart samples were processed by peptic digestion for a mice bioassay. The tissues were homogenized and the resulting samples were subjected to polymerase chain reaction (PCR), which revealed that 64% of them contained the parasite's DNA. The mice bioassay revealed 15 isolates, 8 of them tachyzoites isolates from the peritoneal lavage and 7 from brain cysts. T. gondii genotypes were determined through PCR-RFLP, using the following markers: SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, alt. SAG2, Apico and CS3. Three genotypes were identified, inclued ToxoDB #65, and the other two are not yet described in the literature. Hence, we conclude that the isolates obtained from the metropolitan area of Goiânia showed relatively low genetic diversity. Resumo O objetivo deste estudo foi avaliar as características genotípicas de Toxoplasma gondii isolados de galinhas caipiras da Região Metropolitana de Goiânia, Goiás, Região Centro Oeste do Brasil. A soroprevalência foi de 96% dos animais, determinada por hemaglutinação indireta. As amostras de cérebro e coração foram processadas através da digestão péptica para o bioensaio em camundongos. Os tecidos foram homogeneizados, e as amostras resultantes foram analisadas por reação em cadeia da polimerase (PCR), que possibilitou a detecção do DNA do parasito em 64% deles. Por meio do bioensaio em camundongos, foi possível detectar 15 isolados, 8 deles apresentando taquizoítos na lavagem peritoneal e 7 apresentando cistos cerebrais. A determinação dos genótipos de T. gondii foi realizada por PCR-RFLP com os seguintes marcadores: SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, alt. SAG2, Apico e CS3. Foi possível definir 3 genótipos, incluindo o ToxoDB # 65 e dois deles ainda não foram descritos na literatura. Portanto, conclui-se que os isolados obtidos na região metropolitana de Goiânia apresentaram diversidade genética relativamente baixa.

Background The diagnosis of congenital toxoplasmosis can be inconclusive in many cases. Despite the several serological tests developed, the literature on biomarkers that can assist in the diagnosis of congenital an acute toxoplasmosis is limited. The objective of this study was to analyze the immunoreactive profile of Toxoplasma gondii protein bands with the potential to be biomarkers for diagnosis and prognosis of congenital and acute toxoplasmosis. Methods Peripheral blood samples from women of childbearing age and/or pregnant women diagnosed with acquired toxoplasmosis as well as from congenitally infected children were selected and submitted to immunoblotting for analysis of the immunoreactive bands profile by immunoglobulin G (IgG) antibodies. Results When comparing the immunoreactive bands profile for antibodies present in samples from different groups and subgroups, the 150, 18.5, and 16.96-kDa bands were more immunoreactive with the antibodies present in serum samples from the acquired infection group. The 343, 189, 150, 75, and 42-kDa bands showed more chance to be detected by the symptomatic congenital infection subgroup samples, while the 61, 50, and 16.96-kDa bands were significantly immunoreactive with the acute infection subgroup samples. Conclusions The identification of these potential biomarkers can assist in early diagnosis and treatment of congenital toxoplasmosis.

Toxoplasma gondii can cross the placental barrier, causing fetal infection with potentially severe sequelae. The aim of this study was to evaluate whether the serological screening for toxoplasmosis should be included in the basic neonatal heel prick test in order to establish criteria for the confirmation and/or exclusion of the diagnosis of congenital infection in newborns treated at three public health units in the metropolitan region of Goiania, Goias State, Brazil. Blood samples were collected on filter paper from newborns and later, peripheral blood samples from the mothers and their respective children were obtained to confirm or exclude the diagnosis of suspected congenital infection, by means of an enzyme-linked immunosorbent assay (IgM and IgG) and a polymerase chain reaction assay. From a total of 1,159 blood samples collected on filter paper, 43.92% were reactive to IgG and 0.17% to anti-T. gondii IgM and IgG. One hundred and twenty-seven paired samples (mother and child) were collected following consensual protocols for peripheral blood collection. Results obtained from the filter paper and peripheral blood of the newborns were 90.55% concordant. A comparison of the mother and child blood test results showed agreement regarding the detection of IgG in 90.48% of the samples. The parasite DNA was detected in the peripheral blood of one child. In view of the results obtained in this study, the inclusion of the serological screening for toxoplasmosis in the newborn heel prick test proved to be effective for the early detection of congenital T. gondii infection.

Toxoplasmosis is caused by Toxoplasma gondii and the probability of this infection occurring in the first months of life is usually low because its transmission is related to eating habits. A 6-month-old nursing infant was diagnosed with acute toxoplasmosis, which was identified through anti- T. gondii IgA, IgM and low-avidity IgG serologic assays, polymerase chain reaction (PCR) and mouse bioassay test although its mother was seronegative. This serological divergence between mother and child led us to interview the mother regarding epidemiological factors. During this interview, she reported that she had given her 2-month-old baby a piece of undercooked beef to suck on. After some time, the baby presented fever and cervical lymphadenitis. This report emphasizes the importance of serological surveys of toxoplasmosis in nursing infants presenting with fever and lymphadenitis, in view of the possible acquisition of toxoplasmosis in the first months of life.