909 resultados para Indirect ELISA


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In the present study we evaluated the precision of the ELISA method to quantify caffeine in human plasma and compared the results with those obtained by gas chromatography. A total of 58 samples were analyzed by gas chromatography using a nitrogen-phosphorus detector and routine techniques. For the ELISA test, the samples were diluted to obtain a concentration corresponding to 50% of the absorbance of the standard curve. To determine whether the proximity between the I50 of the standard curve and that of the sample would bring about a more precise result, the samples were divided into three blocks according to the criterion of difference, in modulus, of the I50 of the standard curve and of the I50 of the sample. The samples were classified into three groups. The first was composed of 20 samples with I50 up to 1.5 ng/ml, the second consisted of 21 samples with I50 ranging from 1.51 to 3 ng/ml, and the third of 17 samples with I50 ranging from 3.01 to 13 ng/ml. The determination coefficient (R² = 0.999) showed that the data obtained by gas chromatography represented a reliable basis. The results obtained by ELISA were also reliable, with an estimated Pearson correlation coefficient of 0.82 between the two methods. This coefficient for the different groups (0.88, 0.79 and 0.49 for groups 1, 2 and 3, respectively) showed greater reliability for the test with dilutions closer to I50.

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A vaccine containing crude Toxoplasma gondii rhoptry proteins incorporated in the immunostimulating complexes (ISCOM) adjuvant was tested in pigs for protecting against tissue cyst formation. For this, 38 mixed breed pigs were divided into four groups, G1 (vaccinated challenged, n = 10) received two doses (100 mu g/dose) of the rhoptry vaccine at days 0 and 21, G2 (vaccinated challenged, n = 10) received viable tachyzoites (7 x 10(7)) of the RH strain at day 0, G3 (unvaccinated challenged, n = 10) and G4 (unvaccinated unchallenged, n = 8). Pigs were challenged with 4 x 10(4) VEG strain oocysts 57 days later. The G1 pigs produced high IgG antibody levels in the indirect enzyme-linked immunosorbent assay (ELISA) after the second dose of rhoptry vaccine, but were not clinically protected against a high dose oocyst challenge. Partial protection was observed in G1 at the chronic phase of infection, when compared with G3. Pigs in group 2 developed high antibody levels and were protected against clinic signs. T gondii was not detected in two (G1) and three (G2) pigs by mouse bioassay. The results indicate partial protection in pigs vaccinated with a rhoptry vaccine. (c) 2005 Elsevier B.V. All rights reserved.

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This study evaluated two enzyme-linked immunosorbent assays (ELISA) in the detection of chicken serologic response against Salmonella enterica sorotype Typhimurium. The assays have used as detecting antigen the soluble bacterial proteins of a non-flagellated strain of Salmonella Typhimurium (AgTM), and antibody conjugated to peroxidase or alkaline phosphatase. According to the results, optimal dilutions of antigen (concentration 5.49 mg/mL) and serum samples in both assays were 1:20,000 and 1:1,000, respectively. In such conditions, the ELISA/AgTM was able to detect serological response to Salmonella Typhimurium. Cross-reactions to Salmonella serotypes Gallinarum and Pullorum were seen, but not with other serotypes such as Enteritidis.

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A Erliquiose é uma doença zoonótica causada por bactérias gram-negativas e intracelulares obrigatórias. A Anaplasmose Granulocítica Equina - AGE (anteriormente denominada Erliquiose Granulocítica Equina, EGE) é uma enfermidade sazonal, normalmente auto-limitante em equinos. No Brasil, existem poucos relatos deste agente erliquial, bem como de seus vetores naturais. Atualmente, veterinários têm levantado a suspeita de casos de AGE em equinos com sinais clínicos sugestivos de erliquiose e não responsivos ao tratamento para a piroplasmose equina. O objetivo do presente estudo foi identificar equinos expostos a A. phagocytophilum por meio de técnicas sorológicas e moleculares. Vinte amostras de sangue e soro de equinos da região Centro-oeste do Brasil foram avaliados por meio do exame microscópico de capa leucocitária, ensaio imunoenzimático indireto (ELISA), reação de imunofluorescência indireta (RIFI) e reação em cadeia da polimerase (nested PCR). Adicionalmente, o diagnóstico sorológico de Theileria equi pela RIFI e ELISA foram realizados, assim como o diagnóstico molecular pelo nPCR. Treze (65%) amostras de soro foram positivas para A. phagocytophilum pelo teste de ELISA, entretanto nenhum equino foi positivo pelo exame microscópico da capa leucocitária ou nPCR. Anticorpos IgG anti-T. equi foram detectados em 18 (90%) e 17 (85%) equinos pela RIFI e ELISA, respectivamente e o agente foi detectado em 9 (45%) animais pelo nPCR. Estes dados sugerem importante informação para o entendimento da ocorrência da AGE e piroplasmose equina no Centro-oeste do Brasil.

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This research investigated the pattern of antibody response by means of enzyme linked immunosorbent assay (Elisa) and indirect fluorescent antibody test (IFAT) through the course of experimental Trypanosoma evansi infection in dogs. Clinical and parasitological features were also studied. The average prepatent period was 11.2 days and parasitaemia showed an undulating course. Biometrical study of parasites revealed a mean total length of 21.68mm. The disease was characterized by intermittent fever closely related to the degree of parasitaemia and main clinical signs consisted of pallor of mucous membrane, edema, progressive emaciation and enlargement of palpable lymph nodes. Diagnostic antibody was detected within 12 to 15 days and 15 to 19 days of infection by IFAT and Elisa, respectively. High and persistent antibody levels were detected by both tests and appeared not to correlate with control of parasitaemia

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Doença de Chagas é uma antropozoonose causada por Trypanosoma cruzi que tem os cães como importante reservatório da doença na América do Sul. Este trabalho teve como objetivo avaliar a ocorrência da infecção natural pelo T. cruzi em cães de uma área rural do estado de Mato Grosso do Sul, Brasil. Foram utilizados os testes de imunofluorescência indireta (IFI) e ensaio imunossorvente ligado a enzima (ELISA) em 75 cães residentes na área. Foram detectados anticorpos em 45,3% (n=34) e 24,0% (n=18) nos testes de IFI e ELISA, respectivamente. A real prevalência da infecção foi confirmada como 22,7% (n=17) pelo critério de positividade em ambos os testes. Os resultados obtidos confirmam a infecção chagásica nos cães dessa região.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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The antimutagenic effect of ethanolic extract of propolis (EEP) and honeybee (Apis mellifera) venom, both collected in the State of Sb Paulo, Brazil, was assessed by the Salmonella/microsome assay upon direct- and indirect-acting mutagens. EEP had inhibitory effect (in an ascending order) on the mutagenicity power of daunomycin (TA102), benzo(a)pyrene (TA100), and aflatoxin B-1(TA98) and the venom acted against the mutagenicity of 4-nitro-o-phenylenediamine (TA98) and daunomycin (TA102). (C) 1999 Wiley-Liss, Inc.

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In this work, doped AlGaAs/GaAs parabolic quantum wells (PQW) with different well widths (from 1000 angstrom up to 3000 angstrom) were investigated by means of photoluminescence (PL) measurements. In order to achieve the 2DEG inside the PQW Si delta doping is placed at both side of the well. We have observed that the thickness of this space layer plays a major rule on the characteristics of the 2DEG. It has to be thicker enough to prevent any diffusions of Si to the well and thin enough to allow electrons migration inside the well. From PL measurement, we have observed beside the intra well transitions, indirect transitions involving still trapped electron on the delta doping and holes inside the PQW. For the thinness sample, we have measured a well defined PL peak at low energy side of the GaAs bulk emission. With the increasing of the well thickness this peak intensity decreases and for the thickest sample it almost disappears. Our theoretical calculation indicated that carriers (electron and holes) are more placed at the center of the PQW. In this way, when the well thickness increases the distance between electrons on the delta doping and holes on the well also increases, it decreases the probability of occurrence of these indirect optical transitions. (C) 2012 Published by Elsevier B.V. Selection and/or peer-review under responsibility of Universidade Federal de Juiz de Fora, Brazil.