ABSTRACT Symptomatic forms of toxoplasmosis are a serious public health problem and occur in around 10-20% of the infected people. Aiming to improve the molecular diagnosis of symptomatic toxoplasmosis in Brazilian patients, this study evaluated the performance of real time PCR testing two primer sets (B1 and REP-529) in detecting Toxoplasma gondii DNA. The methodology was assayed in 807 clinical samples with known clinical diagnosis, ELISA, and conventional PCR results in a 9-year period. All samples were from patients with clinical suspicion of several features of toxoplasmosis. According to the minimum detection limit curve (in CT), REP-529 had greater sensitivity to detect T. gondii DNA than B1. Both primer sets were retrospectively evaluated using 515 DNA from different clinical samples. The 122 patients without toxoplasmosis provided high specificity (REP-529, 99.2% and B1, 100%). From the 393 samples with positive ELISA, 146 had clinical diagnosis of toxoplasmosis and positive conventional PCR. REP-529 and B1 sensitivities were 95.9% and 83.6%, respectively. Comparison of REP-529 and B1 performances was further analyzed prospectively in 292 samples. Thus, from a total of 807 DNA analyzed, 217 (26.89%) had positive PCR with, at least one primer set and symptomatic toxoplasmosis confirmed by clinical diagnosis. REP-529 was positive in 97.23%, whereas B1 amplified only 78.80%. After comparing several samples in a Brazilian referral laboratory, this study concluded that REP-529 primer set had better performance than B1 one. These observations were based after using cases with defined clinical diagnosis, ELISA, and conventional PCR.

Abstract: INTRODUCTION Toxoplasmosis during pregnancy can be severe; thus, it is essential to diagnose the disease via serological tests. METHODS An enzyme-linked immunosorbent assay (ELISA) was used to investigate anti-Toxoplasma gondii immunoglobulin A (IgA), M (IgM) and G (IgG) antibodies in 62 high-risk pregnant women. RESULTS Forty-three (69.4%) women were positive for IgA, 31 (50%) for IgG, and 57 (91.9%) for IgM; 4 (6,5%) were positive for IgA but negative for IgM; 10 (16.1%) were negative for IgA and IgM but positive for IgG. CONCLUSIONS Testing for these antibodies can help diagnose infection in pregnant women, thereby contributing to clinical management.

OBJECTIVE:Allergic rhinitis is a disease that affects the upper airways and causes inflammation of the nasal mucosa and it is mediated by IgE antibodies produced after sensitization to environmental allergens. Previous reports have indicated that this disease affects males more often than females. The objective of this study was to verify whether total IgE plasma levels vary between genders in patients suffering from allergic rhinitis.METHODS:A total of 171 adult patients suffering from allergic rhinitis (55 males and 116 females) were enrolled. Total IgE plasma levels were determined using commercial kits, with 140 IU/mL considered as a reference value. The mean total IgE plasma levels were compared according to gender and age.RESULTS:The mean age of the overall patient group with allergic rhinitis was 38.4±19.0 years and a significant difference in age was observed between genders (males: 32.2±17.8 years; females: 41.4±18.9 years; p= 0.0027). Additionally, the mean total IgE plasma levels were higher in males (413.0±143.0 IU/mL) than in females (147.9±98.0 IU/mL) (p<0.0001). These differences persisted even when males and females were stratified by age (up to or older than 20 years of age).CONCLUSIONS:In conclusion, total IgE plasma levels are higher in young adult males than in females suffering from allergic rhinitis. Evaluating total IgE plasma levels can be useful to identify patients at risk of allergic rhinitis in areas with low industrial pollution.

OBJECTIVE: The aim of this study was to verify if ABO phenotypes are associated with allergic rhinitis. METHODS: 168 patients with allergic rhinitis and 168 control individuals from the same geographical region and paired by gender and age were enrolled in the study. ABO phenotypes were identified in red blood cells using the hemagglutination technique. The Fisher exact and chi-squared tests were employed to compare proportions. Statistical significance was set for an alpha error of 5% (p-value < 0.05). RESULTS: The overall differences in the frequencies of the ABO phenotypes of patients and controls were marginal (χ2: 7.569; degrees of freedom (DF): 3; p-value = 0.055) however the O blood group was associated with allergic rhinitis (χ2: 5.764; DF: 1; p-value = 0.016; OR: 1.735; CI 95%: 1.127-2.673). The differences in the frequencies of the O phenotype in patients and controls were statistically different for men (χ2: 8.520; DF: 1; p-value = 0.003) but not for women (χ2: 0.6375; DF: 1; p-value = 0.4246). The A phenotype was associated with protection (OR: 0.4385; CI 95%: 0.2043-0.9415; p-value = 0.049) and the O phenotype was associated with susceptibility (OR = 2.789; CI 95%: 1.385-5.616; p-value = 0.005) to allergic rhinitis only for men. CONCLUSION: The O blood group phenotype is associated with allergic rhinitis in male but not in female patients.

CONTEXTO E OBJETIVOS: A toxoplasmose, quando transmitida durante a gestação, pode causar graves sequelas em fetos e neonatos. Anticorpos maternos podem ser indicadores de risco ou de imunidade. O objetivo foi avaliar a positividade dos anticorpos das classes imunoglobulina M (IgM) e imunoglobulina G (IgG) anti-Toxoplasma gondii (anti-T. gondii), bem como a avidez de IgG em gestantes e seus neonatos. TIPO DE ESTUDO E LOCAL: Estudo transversal em ambulatório de gestação de alto risco. MÉTODOS: Anticorpos anti-T. gondii foram avaliados em amostras de soro de gestantes (n = 87) e seus respectivos neonatos (n = 87) com o uso dos métodos imunofluorescência indireta (IFI) (IgM e IgG), ensaio imunoenzimático (ELISA) (IgG) e avidez. RESULTADOS: Anticorpos anti-T. gondii foram identificados em 64,4% das amostras de soro das mães e seus bebês (56/87). Com exceção de duas amostras de soro materno (2,3%), todas as demais foram negativas anticorpos IgM anti-T. gondii determinado pela IFI. Os resultados mostraram que 92,9% das gestantes tinham índices elevados de avidez de IgG (> 30%) e 4 amostras apresentaram índices de avidez entre 16-30%. Duas gestantes no terceiro trimestre da gravidez eram positivas IgM anti-T. gondii; seus bebês apresentaram índices de avidez entre 16 e 30%. Os índices de avidez dos soros dos outros 83 recém-nascidos foram semelhantes àqueles encontrados nas amostras maternas. CONCLUSÕES: Os resultados mostraram que 2% das gestantes estavam sob risco de transmissão de T. gondii durante o período gestacional. Estes dados parecem refletir a real situação da toxoplasmose gestacional na região noroeste do Estado de São Paulo.

CONTEXT AND OBJECTIVE: Toxoplasmosis transmission during pregnancy can cause severe sequelae in fetuses and newborns. Maternal antibodies may be indicators of risk or immunity. The aim here was to evaluate seropositivity for anti-Toxoplasma gondii (anti-T. gondii) immunoglobulin M (IgM) and immunoglobulin G (IgG) antibodies and IgG avidity in pregnant women and their newborn infants. DESIGN AND SETTING: Cross-sectional study in a high-risk pregnancy outpatient clinic. METHODS: Serum samples from pregnant women (n = 87) and their respective newborns (n = 87) were evaluated for anti-T. gondii antibodies using indirect immunofluorescence (IIF) (IgM and IgG), enzyme-linked immunosorbent assay (ELISA) (IgG) and an avidity test. RESULTS: Anti-T. gondii antibodies were identified in 64.4% of the serum samples from the mothers and their infants (56/87). Except for two maternal serum samples (2.3%), all others were negative for anti-T. gondii IgM antibodies, using IIF. The results showed that 92.9% of the pregnant women had high IgG avidity indexes (> 30%) and four samples had avidity indexes between 16 and 30%. Two women in the third trimester of pregnancy were positive for anti-T. gondii IgM antibodies; their babies had avidity indexes between 16 and 30%. The avidity indexes of serum from the other 83 newborns were similar to the results from their mothers. CONCLUSIONS: The results showed that 2% of the pregnant women were at risk of T. gondii transmission during the gestational period. These data seem to reflect the real situation of gestational toxoplasmosis in the northwestern region of the state of São Paulo.

OBJECTIVE: Adenosine deaminase acts on adenosine and deoxyadenosine metabolism and modulates the immune response. The adenosine deaminase G22A polymorphism (20q.11.33) influences the level of adenosine deaminase enzyme expression, which seems to play a key role in maintaining pregnancy. The adenosine deaminase 2 phenotype has been associated with a protective effect against recurrent spontaneous abortions in European Caucasian women. The aim of this study was to investigate whether the G22A polymorphism of the adenosine deaminase gene is associated with recurrent spontaneous abortions in Brazilian women. METHODS: A total of 311 women were recruited to form two groups: G1, with a history of recurrent spontaneous abortions (N = 129), and G2, without a history of abortions (N = 182). Genomic DNA was extracted from peripheral blood with a commercial kit and PCR-RFLP analysis was used to identify the G22A genetic polymorphism. Fisher's exact test and odds ratio values were used to compare the proportions of adenosine deaminase genotypes and alleles between women with and without a history of recurrent spontaneous abortion (p<0.05). The differences between mean values for categorical data were calculated using unpaired t tests. The Hardy-Weinberg equilibrium was assessed with a chi-square test. RESULTS: Statistically significant differences were identified for the frequencies of adenosine deaminase genotypes and alleles between the G1 and G2 groups when adjusted for maternal age. CONCLUSIONS:The results suggest that the adenosine deaminase *2 allele is associated with a low risk for recurrent spontaneous abortions, but this association is dependent on older age.

Infectious and parasitic diseases affecting women during their reproductive age may result in vertical transmission. The aim of this study was to determine the seroprevalence for TORSCH among pregnant women receiving care at a university hospital. Records of 574 pregnant women who received medical attention from January 2006 to December 2007 were assessed. The mean age was 27.2 ± 6.5 years ranging from 13 to 44. The results of the immunodiagnostic tests were: 62.0% (345/556) for IgG and 3.4% (19/556) for IgM anti-T. gondii; 93.1% (433/465) for IgG and 0.6% (3/465) for IgM anti-rubella; 0.9% (5/561) for VDRL; 1.8% (10/554) for HBsAg; 0.7% (4/545) for anti-HCV and 2.1% (11/531) for HIV. In conclusion, the results of immunodiagnostic tests for the TORSCH panel among pregnant women attending a perinatal service of a university hospital are in agreement with those reported by previous studies and by governmental sources.