Dot-dye-immunoassay for the diagnosis of schistosomiasis mansoni

A new serological assay dot-dye-immunoassay (dot-DIA) was evaluated for the diagnosis of schistosomiasis mansoni. This method consist of four steps: (a) biding of antigens to a nitrocellulose membrane (NC); (b) blocking of free sites of the NC; (c) incubation in specific primary antibody; (d) detection of primary antibody reactivity by color development using second antibody coupled to textile dyes. Sera from 82 individuals, 61 with Schistosoma mansoni eggs in the stool and 21 stool negative were tested by ELISA, dot-ELISA, and dotDIA. A high level of agreement between the methods tested was observed for all sera tested: ELISA x dot-ELISA: 95.1%, ELISA x dot-DIA: 92.7% and dot-ELISA x dot-DIA: 97.6%. In this study, dot-DIA proved to be a feasible, sensitive, rapid and practical test for the diagnosis of shcistosomiasis.

Coeficiente de cultura da alface hidropônica baseado no conceito de graus-dia

O objetivo principal deste trabalho foi propor a adoção de um coeficiente de cultura térmico, com a aplicação do conceito de graus-dia acumulados após o transplantio, para estimativa da evapotranspiração diária (ETc) dos cultivares de alface Grand Rapids, Regina e Great Lakes em sistema hidropônico do tipo NFT ("nutrient film technique"), em ambiente protegido. Posteriormente, o modelo matemático ajustado ãos valores observados de Kc (coeficiente de cultura) foi utilizado para a estimativa de ETc no período pós-transplantio por meio dos métodos: Penman-Monteith parametrizado pela FÃO, em 1998 (PMF); Penman-Monteith modificado para condições aerodinâmicas, características de ambientes protegidos (PMAP); Penman-Monteith para ambiente protegido simplificado (PMAPS); Priestley-Taylor (PT); Radiação solar (RS); e Radiação solar simplificado (RSS). Verificou-se que o modelo sigmoidal com quatro parâmetros ajustou-se bem aos valores observados de Kc, proporcionando coeficientes de determinação ajustados de 0,99, 0,98 e 0,98 para os cultivares Grand Rapids, Regina e Great Lakes, respectivamente. No que se refere às estimativas diárias de ETc, os métodos PMF, PMAP e RS apresentaram melhor desempenho estatístico em relagão aos métodos PMAPS, PT e RSS, independentemente do cultivar avaliado. No entanto, o método PMF subestimou sistematicamente a ETc no período inicial de crescimento da alface.

Imunidade relativa à poliomielite em crianças de zero a dez anos, após o "Quarto Dia Nacional de Vacinação Contra a Poliomielite" com a vacinação oral trivalente, tipo Sabin, em área da Região da Grande São Paulo, SP (Brasil), 1982

Efetuou-se um levantamento soro-epidemiológico, abrangendo 898 crianças, para a verificação da imunidade aos poliovírus 1, 2 e 3 por meio da determinação dos títulos de anticorpos neutralizantes. De 522 crianças, ou seja, de 58,1% da população estudada, foi possível a obtenção de informações referentes à situação vacinal a partir de anotações existentes nas próprias Cadernetas de Vacinação. Neste último sub-grupo foi estudado o estado imunitário em relação ao número de doses de vacina oral tipo Sabin recebidas. Os resultados mostraram alta proporção de imunes para os três tipos de poliovírus entre as crianças estudadas, porém também revelaram a existência de lacuna imunitária nos menores de um ano, especialmente no primeiro semestre de vida. Não foram observadas diferenças importantes na proporção de imunes segundo o sorotipo considerado. Salientou-se a necessidade de um alerta constante e permanente em relação à ocorrência de possíveis surtos epidêmicos em segmentos mais vulneráveis da população, particularmente entre crianças menores de 1 ano.

Merenda no dia alimentar de crianças matriculadas em Centros de Educação e Alimentação do Pré-Escolar

Estudou-se o papel da merenda no comportamento alimentar de 346 pré-escolares (PE) matriculados em Centros de Educação e Alimentação do Pré-Escolar (CEAPEs) de seis municípios do Estado de São Paulo, Brasil. Em entrevista domiciliar com a mãe ou responsável pela alimentação do PE, obteve-se o número e a quantidade dos alimentos ingeridos pela criança antes e após sua participação no CEAPE. Verificou-se que essa refeição escolar interfere tanto na quantidade dos alimentos consumidos quanto no número das refeições diárias feitas no lar. Os resultados mostraram que 178 (51,4%) PE reduziram a ingestão alimentar de casa, mediante exclusão de refeições e/ou diminuição da quantidade de alimentos habitualmente ingerida, após receberem a merenda no CEAPE. Destas, 115 (64,6%) apresentaram dieta insuficiente em energia; 48 (13,9%) aumentaram a quantidade dos alimentos habituais e/ou incluíram refeições. Ainda assim, 23 (47,9%) apresentaram consumo energético deficiente. Entre os 120 (34,7%) que não tiveram nenhuma modificação em seu dia alimentar, 61 (51,7%) mostraram ingestão calórica inadequada. Concluiu-se ser necessário orientar a família sobre o papel da merenda como suplemento alimentar e não como substituto de refeições no lar.

Hospitalizações por infarto agudo do miocárdio segundo o dia da semana: estudo retrospectivo

OBJETIVO: A literatura tem relatado a associação do infarto com a segunda-feira, sugerindo a existência de mecanismos de "gatilho" no desencadeamento da doença. Foi realizado estudo para verificar a distribuição semanal das hospitalizações por infarto do miocárdio e sua associação ao sexo e à categoria da internação (condição social) na região de Ribeirão Preto de 1987 a 1996. MÉTODOS: Foram estudadas 173.982 hospitalizações por doenças cardíacas e vasculares cerebrais, incluindo 5.804 casos de infarto agudo do miocárdio, referidos a um centro de registro contínuo de informações de assistência hospitalar. As hospitalizações foram classificadas em particulares, de sistemas privados de pré-pagamento e do Sistema Único de Saúde (SUS). Foram estudadas as associações com o sexo, a condição do egresso e a categoria da internação. RESULTADOS/CONCLUSÕES: A distribuição semanal de hospitalizações por doenças cardiovasculares e infarto agudo do miocárdio apresentou um pico às segundas-feiras: 19,3% e 16,9% dos casos, respectivamente. Houve queda acentuada no final de semana (8,6%) dos casos de doenças cardiovasculares e menor redução dos casos de infarto agudo do miocárdio (12,7%), sendo essas diferenças estatisticamente significantes. Houve diferenças nos padrões de distribuição semanal dos pacientes do SUS (4.120), dos sistemas de pré-pagamento (1.225) e dos particulares (459). Para os pacientes do SUS e dos sistemas de pré-pagamento o número de hospitalizações foi elevado no início da semana e houve uma diminuição gradual no restante dos dias. Para os particulares, o número de hospitalizações foi baixa no domingo e houve elevação na sexta e sábado. Não houve associação com o sexo ou com a condição de saída.

DOT-ELISA for detection of anti-Cysticercus cellulosae antibodies in human cerebrospinal fluid using a new solid-phase (resin-treated polyester fabrics): preliminary report

A dot enzyme-linked immunosorbent assay (DOT-ELISA) was developed to detect specific antibodies in cerebrospinal fluid (CSF) for human neurocysticercosis immunodiagnosis, with Cysticercus cellulosae antigen dotted on a new solid-phase. This was represented by sheets of a synthetic polyester fabric impregnated with a polymerized resin (N-methylol-acrylamide). A very stable preparation was thus obtained, the antigen being covalently bound by cross-linking with free N-methylol groups on the resin. Since robust, no special care was necessary for handling the solid-phase. The test could be performed at room-temperature. From 30 CSF samples assayed, 14 were positive, from a group of 15 cases of neurocysticercosis, with titers from 1 to 128; 15 other samples, from normals or other neurological diseases, were all negative. Test characteristics seem to indicate it as adequate for epidemiological surveys. A more detailed study on sensitivity, specificity, reproducibility and the use in serum samples is being conducted.

Performance indexes of a dot-enzime-linked immunosorbent assay (dot-ELISA) and an ezyme-linked immunosorbent assay (IgG-ELISA) for field surveys of new world leishmaniasis

Diagnostic performance indexes of sensitivity, specificity, positive predictive value and efficiency were determined for dot-ELISA and IgG-ELISA tests in 340 leishmaniasis sera. Sensitivity of the dot-ELISA was significantly lower than IgG-ELISA's; the two tests had indexes of specificity and positive predictive value of the same magnitude. Seventy-eight sera gave a negative dot-ELISA test result and a positive IgG-ELISA test result. When sera were classified according to different criteria as how to interpret this diversity, the kappa statistic did not corroborate the classification indicating that the two tests display a substantial strength of agreement. The results presented indicate that performance indexes accrued in a survey where variables arc well known may be extrapolated to other population studies if the disease presents itself as highly prevalent (due to a selection bias or not) and may be expected to discriminate a disease status among test positives.

Standardization of dot-ELISA for the serological diagnosis of toxocariasis and comparision of the assay with ELISA

The dot-enzyme-linked immunosorbent assay (dot-ELISA) was standardized using somatic (S) and excretory-secretory (ES) antigens of Toxocara-canis for the detection of specific antibodies in 22 serum samples from children aged 1 to 15 years, with clinical signs of toxocariasis. Fourteen serum samples from apparently normal individuals and 28 sera from patients with other pathologies were used as controls. All samples were used before and after absorption with Ascaris suum extract. When the results were evaluated in comparison with ELISA, the two tests were found to have similar sensitivity, but dot-ELISA was found to be more specific in the presence of the two antigens studied. Dot-ELISA proved to be effective for the diagnosis of human toxocariasis, presenting advantages in terms of yield, stability, time and ease of execution and low cost.

Measles serodiagnosis: standardization and evaluation of a dot-ELISA

A Dot-ELISA using a measles virus (MV) antigen obtained by sodium deoxycholate treatment was standardized and evaluated for IgM and IgG antibody detection in measles patients and measles-vaccinated subjects. A total of 192 serum samples were studied, comprising 47 from patients with acute and convalescent measles, 55 from 9-month old children prior to measles vaccination and 41 from children of the same age after vaccination, and 49 from patients with unrelated diseases. The diagnostic performances of the IgG Dot-ELISA and IgG immuno fluorescence test (IFT) were found to be close, varying from 0.97 to 1.00 in sensitivity and the specificities were maximum (1.00). Nevertheless, the sensitivity of the IgM Dot-ELISA (0.85) was higher than that (0.63) of the IgM IFT, although both assays had comparably high (1.00) specificities. The IgM Dot-ELISA in particular proved to be more sensitive in relation to other assays studied by revealing antibodies in 80.0% (12/15) of vaccinated children on the 15th day after immunization. In contrast the IgM IFT, failed to detect antibodies in the same group of vaccinated children. The stability of the MV antigen was longer than that of the IFT antigen, and the reproducibility of the Dot-Elisa was satisfactory.

Alkaline soluble Trypanosoma cruzi epimastigote antigen (ASEA) applied to dot-ELISA

The alkaline soluble Trypanosoma cruzi epimastigote antigen (ASEA) was assessed in dot-ELISA for the diagnosis of Chagas' disease. Serum samples (355) from chagasic and non-chagasic patients were studied, and IgG antibodies to ASEA were found in all patients with chronic Chagas' disease. In non-chagasic patients 95.6% were negative, except for those with leishmaniasis (visceral and mucocutaneous), and some patients from control group reacted in low titers. The data indicate that dot-ELISA using ASEA is suitable for seroepidemiologic surveys to be employed in endemic areas for Chagas' disease.

Dot-ELISA-IgM in saliva for the diagnosis of human leptospirosis using polyester fabric-resin as support (Preliminary Report)

In order to improve the diagnosis of human leptospirosis, we standardized the dot-ELISA for the search of specific IgM antibodies in saliva. Saliva and serum samples were collected simultaneously from 20 patients with the icterohemorrhagic form of the disease, from 10 patients with other pathologies and from 5 negative controls. Leptospires of serovars icterohaemorrhagiae, canicola, hebdomadis, brasiliensis and cynopteri grown in EMJH medium and mixed together in equal volumes, were used as antigen at individual protein concentration of 0.2 µg/µl. In the solid phase of the test we used polyester fabric impregnated with N-methylolacrylamide resin. The antigen volume for each test was 1µl, the saliva volume was 8 µl, and the volume of peroxidase-labelled anti-human IgM conjugate was 30 µl. A visual reading was taken after development in freshly prepared chromogen solution. In contrast to the classic nitrocellulose membrane support, the fabric support is easy to obtain and to handle. Saliva can be collected directly onto the support, a fact that facilitates the method and reduces the expenses and risks related to blood processing.

Dot-enzyme-linked immunosorbent assay (Dot-ELISA) for detection of pneumococcal polysaccharide antigens in pleural fluid effusion samples.: Comparison with bacterial culture, counterimmunoelectrophoresis and latex agglutination

A dot-enzyme-linked immunosorbent assay (Dot-ELISA) for pneumococcal antigen detection was standardized in view of the need for a rapid and accurate immunodiagnosis of acute pneumococcal pneumonia. A total of 442 pleural fluid effusion samples (PFES) from children with clinical and laboratory diagnoses of acute bacterial pneumonia, plus 38 control PFES from tuberculosis patients and 20 negative control serum samples from healthy children were evaluated by Dot-ELISA. The samples were previously treated with 0.1 M EDTA pH 7.5 at 90°C for 10 min and dotted on nitrocellulose membrane. Pneumococcal omniserum diluted at 1:200 was employed in this assay for antigen detection. When compared with standard bacterial culture, counterimmunoelectrophoresis and latex agglutination techniques, the Dot-ELISA results showed relative indices of 0.940 to sensitivity, 0.830 to specificity and 0.760 to agreement. Pneumococcal omniserum proved to be an optimal polyvalent antiserum for the detection of pneumococcal antigen by Dot-ELISA. Dot-ELISA proved to be a practical alternative technique for the diagnosis of pneumococcal pneumonia.

Dot-ELISA for the detection of IgM and IgG antibodies to Schistosoma mansoni worm and egg antigens, associated with egg excretion by patients

Human schistosomiasis, caused by Schistosoma mansoni, is highly prevalent in Brazil and usually diagnosed by time consuming stool analysis. Serological tests are of limited use in this disease, mainly for epidemiological studies, showing no discrimination between previous contact with the parasite and active infections. In the present study, we standardized and compared a Dot-ELISA for IgM and IgG antibodies against S. mansoni antigens from eggs and worms with a routine IgG and IgM immunofluorescence assay using similar antigens, in the study of sera from 27 patients who had quantified egg stool excretion. The positivity obtained for IgG Dot-ELISA was 96.3% and 88.9% for IgM Dot-ELISA with worm antigen and 92.6% and 90.9% with egg antigen. The IFI presented similar positivities using worm antigen, 92.6% (IgG) and 96.3% (IgM),and lower results with egg antigen, 77.8% (IgG and IgM). The patients studied were divided into two groups according to their egg excretion, with greater positivity of serological tests in higher egg excreters. When comparing the quantitative egg excretion and the serological titers of the patients, we detected a correlation only with IgM Dot-ELISA, with r=0.552 (p=0.0127). These data show that Dot-ELISA can be used for the detection of specific antibodies against S. mansoni in sera from suspected patients or in epidemiological studies and, with further purification of egg antigen and larger samples, IgM Dot-ELISA could be a possible tool for rough estimates of parasite burden in epidemiological studies.