RESUMEN
Our objective was to evaluate the diagnosis of swine cysticercosis by examining "ante mortem" (inspection of the tongue), "post mortem" (inspection and detailed necropsy) and ELISA for research in serum of antibodies (Ab-ELISA) and antigens (Ag-ELISA). Seven (7) pigs were experimentally infected orally with eggs of Taenia solium and another 10 were naturally infected. In the pigs experimentally infected, inspection of the tongue was negative in all animals, in the routine inspection detailed necropsy and cysticercis were identified in all of them. In pigs with heavy natural infection, inspection of the tongue identified cysticerci in two (20%), while at inspection with necropsy the parasites were identified in large quantities in all animals. In ELISA for antibody search (Ab-ELISA) TS-14 recombinant protein was used, and in search for antigen (Ag-ELISA) a monoclonal antibody against this protein. In animals experimentally infected, blood was collected weekly for 140 days. The Ab-ELISA identified an increase in titers of antibody to cysticerci 21 days after infection, and at the end of the experimental period six animals (86%) were positive to the test. The search for circulating antigens (Ag-ELISA) was positive in two pigs 28 to 91 days after infection. All naturally infected pigs were positive for Ag-ELISA and Ab-ELISA. The search for antibodies and antigens by ELISA in serum from 30 pigs of a local farm and without history of cysticercosis was negative. Thus, the use of TS-14 antigen in ELISA test (Ab-ELISA) can be useful for the diagnosis of cysticercosis in pigs with low infection.(AU)
Nosso objetivo foi avaliar o diagnóstico de cisticercose suína através do exame "ante mortem" (inspeção da língua), "post mortem" (inspeção e necropsia detalhada) e teste de ELISA para a pesquisa no soro de anticorpos (Ab-ELISA) e antígenos (Ag -ELISA). Sete (7) suínos foram infectados experimentalmente por via oral com ovos de Taenia solium e outros 10 eram portadores de infecção natural generalizada. Nos suínos experimentalmente infectados, a inspeção da língua foi negativa em todos os animais, na inspeção 4 (57%) estavam infectados, a necropsia detalhada identificou cisticercos em todos os animais. Nos animais com infecção natural generalizada, a inspeção da língua identificou cisticercos em 2 (20%), enquanto que a inspeção e a necropsia os parasitas foram identificados em grande quantidade em todos os animais. No teste de ELISA para a pesquisa de anticorpos (Ab-ELISA) foi utilizada a proteína recombinante TS-14 e para a pesquisa de antígenos (Ag-ELISA) um anticorpo monoclonal produzido contra esta proteína. Nos animais experimentalmente infectados o sangue foi coletado semanalmente por um período de 140 dias. O Ab-ELISA identificou um aumento nos títulos de anticorpos para cisticercos 21 dias após a infecção, sendo que no final do período experimental 6 animais (86%) foram positivos ao teste. A pesquisa de antígenos circulantes (Ag-ELISA), foi positiva em 2 animais, entre os dias 21 e 91 após a infecção . Todos os suínos com infecção natural generalizada foram positivos para Ag-ELISA e Ab-ELISA.A pesquisa de anticorpos e antígenos pelo ELISA realizada no soro de 30 suínos procedentes de uma criação local sem historia de cisticercose foi negativa. Assim o uso do antígeno TS-14 (Ac-ELISA), pode ser útil para o diagnóstico da cisticercose em suínos com baixa infecção.(AU)
Asunto(s)
Animales , Porcinos/parasitología , Cisticercosis/veterinaria , Cisticercosis/diagnóstico , Taenia solium/patogenicidad , Autopsia , Ensayo de Inmunoadsorción Enzimática , Lengua/fisiopatología , Cysticercus/inmunologíaRESUMEN
Our objective was to evaluate the diagnosis of swine cysticercosis by examining "ante mortem" (inspection of the tongue), "post mortem" (inspection and detailed necropsy) and ELISA for research in serum of antibodies (Ab-ELISA) and antigens (Ag-ELISA). Seven (7) pigs were experimentally infected orally with eggs of Taenia solium and another 10 were naturally infected. In the pigs experimentally infected, inspection of the tongue was negative in all animals, in the routine inspection detailed necropsy and cysticercis were identified in all of them. In pigs with heavy natural infection, inspection of the tongue identified cysticerci in two (20%), while at inspection with necropsy the parasites were identified in large quantities in all animals. In ELISA for antibody search (Ab-ELISA) TS-14 recombinant protein was used, and in search for antigen (Ag-ELISA) a monoclonal antibody against this protein. In animals experimentally infected, blood was collected weekly for 140 days. The Ab-ELISA identified an increase in titers of antibody to cysticerci 21 days after infection, and at the end of the experimental period six animals (86%) were positive to the test. The search for circulating antigens (Ag-ELISA) was positive in two pigs 28 to 91 days after infection. All naturally infected pigs were positive for Ag-ELISA and Ab-ELISA. The search for antibodies and antigens by ELISA in serum from 30 pigs of a local farm and without history of cysticercosis was negative. Thus, the use of TS-14 antigen in ELISA test (Ab-ELISA) can be useful for the diagnosis of cysticercosis in pigs with low infection.
Nosso objetivo foi avaliar o diagnóstico de cisticercose suína através do exame "ante mortem" (inspeção da língua), "post mortem" (inspeção e necropsia detalhada) e teste de ELISA para a pesquisa no soro de anticorpos (Ab-ELISA) e antígenos (Ag -ELISA). Sete (7) suínos foram infectados experimentalmente por via oral com ovos de Taenia solium e outros 10 eram portadores de infecção natural generalizada. Nos suínos experimentalmente infectados, a inspeção da língua foi negativa em todos os animais, na inspeção 4 (57%) estavam infectados, a necropsia detalhada identificou cisticercos em todos os animais. Nos animais com infecção natural generalizada, a inspeção da língua identificou cisticercos em 2 (20%), enquanto que a inspeção e a necropsia os parasitas foram identificados em grande quantidade em todos os animais. No teste de ELISA para a pesquisa de anticorpos (Ab-ELISA) foi utilizada a proteína recombinante TS-14 e para a pesquisa de antígenos (Ag-ELISA) um anticorpo monoclonal produzido contra esta proteína. Nos animais experimentalmente infectados o sangue foi coletado semanalmente por um período de 140 dias. O Ab-ELISA identificou um aumento nos títulos de anticorpos para cisticercos 21 dias após a infecção, sendo que no final do período experimental 6 animais (86%) foram positivos ao teste. A pesquisa de antígenos circulantes (Ag-ELISA), foi positiva em 2 animais, entre os dias 21 e 91 após a infecção . Todos os suínos com infecção natural generalizada foram positivos para Ag-ELISA e Ab-ELISA.A pesquisa de anticorpos e antígenos pelo ELISA realizada no soro de 30 suínos procedentes de uma criação local sem historia de cisticercose foi negativa. Assim o uso do antígeno TS-14 (Ac-ELISA), pode ser útil para o diagnóstico da cisticercose em suínos com baixa infecção.
Asunto(s)
Animales , Autopsia , Cisticercosis/diagnóstico , Cisticercosis/veterinaria , Ensayo de Inmunoadsorción Enzimática , Porcinos/parasitología , Taenia solium/patogenicidad , Cysticercus/inmunología , Lengua/fisiopatologíaRESUMEN
The Apical Membrane Antigen 1 (AMA-1) is considered a promising candidate for development of a malaria vaccine against asexual stages of Plasmodium. We recently identified domain II (DII) of Plasmodium vivax AMA-1 (PvAMA-1) as a highly immunogenic region recognised by IgG antibodies present in many individuals during patent infection with P. vivax. The present study was designed to evaluate the immunogenic properties of a bacterial recombinant protein containing PvAMA-1 DII. To accomplish this, the recombinant protein was administered to mice in the presence of each of the following six adjuvants: Complete/Incomplete Freund's Adjuvant (CFA/IFA), aluminium hydroxide (Alum), Quil A, QS21 saponin, CpG-ODN 1826 and TiterMax. We found that recombinant DII was highly immunogenic in BALB/c mice when administered in the presence of any of the tested adjuvants. Importantly, we show that DII-specific antibodies recognised the native AMA-1 protein expressed on the surface of P. vivax merozoites isolated from the blood of infected patients. These results demonstrate that a recombinant protein containing PvAMA-1 DII is immunogenic when administered in different adjuvant formulations, and indicate that this region of the AMA-1 protein should continue to be evaluated as part of a subunit vaccine against vivax malaria.
Asunto(s)
Anticuerpos Antiprotozoarios/sangre , Antígenos de Protozoos/inmunología , Vacunas contra la Malaria/inmunología , Proteínas de la Membrana/inmunología , Proteínas Protozoarias/inmunología , Adyuvantes Inmunológicos/farmacología , Animales , Anticuerpos Monoclonales/inmunología , Formación de Anticuerpos , Femenino , Ratones , Ratones Endogámicos BALB C , Plasmodium vivax/inmunología , Proteínas Recombinantes/inmunología , Vacunas de Subunidad/inmunologíaRESUMEN
Glycoproteins from the total vesicular fluid of Taenia crassiceps (VF-Tc) were prepared using three different purification methods, consisting of ConA-lectin affinity chromatography (ConA-Tc), preparative electrophoresis (SDS-PAGE) (14 gp-Tc), and monoclonal antibody immunoaffinity chromatography (18/14-Tc). The complex composition represented by the VF-Tc and ConA-Tc antigens revealed peptides ranging from 101- to 14-kDa and from 92- to 12-kDa, respectively. Immunoblotting using lectins confirmed glucose/mannose (glc/man) residues in the 18- and 14-kDa peptides, which are considered specific and immunodominant for the diagnosis of cysticercosis, and indicated that these fractions are glycoproteins. Serum antibodies from a patient with neurocysticercosis that reacted to the 14 gp band from T. crassiceps (Tc) were eluted from immunoblotting membranes and showed reactivity to 14 gp from Taenia solium. In order to determine the similar peptide sequence, the N-terminal amino acid was determined and analyzed with sequences available in public databases. This sequence revealed partial homology between T. crassiceps and T. solium peptides. In addition, mass spectrometry along with theoretical M(r) and pI of the 14 gp-Tc point suggested a close relationship to some peptides of a 150-kDa protein complex of the T. solium previously described. The identification of these common immunogenic sites will contribute to future efforts to develop recombinant antigens and synthetic peptides for immunological assays.
Asunto(s)
Antígenos Helmínticos/química , Glicoproteínas/química , Proteínas del Helminto/química , Taenia/inmunología , Secuencia de Aminoácidos , Animales , Anticuerpos Monoclonales/inmunología , Especificidad de Anticuerpos , Antígenos Helmínticos/inmunología , Western Blotting , Cromatografía de Afinidad , Reacciones Cruzadas , Cysticercus/inmunología , Electroforesis en Gel de Poliacrilamida , Femenino , Glicoproteínas/inmunología , Proteínas del Helminto/inmunología , Lectinas , Ratones , Ratones Endogámicos BALB C , Datos de Secuencia Molecular , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Taenia solium/inmunologíaRESUMEN
The interactions between three different protein antigens and dioctadecyldimethylammonium bromide (DODAB) dispersed in aqueous solutions from probe sonication or adsorbed as one bilayer onto particles was comparatively investigated. The three model proteins were bovine serum albumin (BSA), purified 18 kDa/14 kDa antigens from Taenia crassiceps (18/14-Tcra) and a recombinant, heat-shock protein hsp-18 kDa from Mycobacterium leprae. Protein-DODAB complexes in water solution were characterized by dynamic light scattering for sizing and zeta-potential analysis. Cationic complexes (80-100 nm of mean hydrodynamic diameter) displayed sizes similar to those of DODAB bilayer fragments (BF) in aqueous solution and good colloid stability over a range of DODAB and protein concentrations. The amount of cationic lipid required for attaining zero of zeta-potential at a given protein amount depended on protein nature being smaller for 18 kDa/14 kDa antigens than for BSA. Mean diameters for DODAB/protein complexes increased, whereas zeta-potentials decreased with NaCl or protein concentration. In mice, weak IgG production but significant cellular immune responses were induced by the complexes in comparison to antigens alone or carried by aluminum hydroxide as shown from IgG in serum determined by ELISA, delayed type hypersensitivity reaction from footpad swelling tests and cytokines analysis. The novel cationic adjuvant/protein complexes revealed good colloid stability and potential for vaccine design at a reduced DODAB concentration.
Asunto(s)
Adyuvantes Inmunológicos/química , Lípidos/química , Compuestos de Amonio Cuaternario/química , Animales , Antígenos Bacterianos/inmunología , Antígenos Helmínticos/inmunología , Cationes/química , Cationes/inmunología , Bovinos , Células Cultivadas , Química Farmacéutica , Citocinas/análisis , Estabilidad de Medicamentos , Ensayo de Inmunoadsorción Enzimática , Femenino , Hipersensibilidad Tardía/inmunología , Lípidos/inmunología , Ratones , Ratones Endogámicos BALB C , Mycobacterium leprae/inmunología , Nanopartículas , Tamaño de la Partícula , Compuestos de Amonio Cuaternario/inmunología , Albúmina Sérica Bovina/inmunología , Taenia/inmunologíaRESUMEN
Sera from 88 patients from Santa Catarina and São Paulo states of Brazil, with epileptic seizures who underwent cerebral computed tomography (CT) were analyzed for the detection of antibodies to T. solium cysticercus by ELISA and Immunoblot (IB) with the following antigens: Taenia solium cysticercus total saline (Tso), Taenia crassiceps cysticercus vesicular fluid (Tcra-vf) and T. crassiceps cysticercus glycoproteins (Tcra-gp). ELISA carried out with Tso, Tcra-vf and Tcra-gp antigens showed 95 percent, 90 percent and 80 percent sensitivities, respectively, and 68 percent, 85 percent and 93 percent specificities, respectively. In the epileptic patients group, ELISA positivity was 30 percent, 51 percent and 35 percent with Tso, Tcra-vf and Tcra-gp antigens respectively. Considering the IB as the confirmatory test, the positivity was 16 percent (14/88) in the epileptic patients total group and 22 percent (12/54) in the epileptic patients with positive CT and signals of cysticercosis. We found a significant statistical correlation among ELISA or IB results and the phase of the disease when any antigens were used (p < 0.05). We emphasize the need to introduce in the laboratory routine the search for neurocysticercosis (NC) in patients presenting with epileptic seizures because of the high risk of acquiring NC in our region and its potential cause of epilepsy.
Amostras de soro de 88 pacientes dos Estados de Santa Catarina e São Paulo, Brasil, com crises epilépticas e que se submeteram a exame de Tomografia Computadorizada (TC), foram examinadas para detecção de anticorpos anti-cisticercos de Taenia solium por meio de ELISA e Immunoblot (IB) utilizando-se os seguintes antígenos: extrato salino total de cisticercos de T. solium (Tso); líquido vesicular de Taenia crassiceps (Tcra-vf) e glicoproteínas purificadas de cisticercos de T. crassiceps (Tcra-gp). Os resultados de ELISA com os antígenos Tso, Tcra-vf e Tcra-gp mostraram 95 por cento, 90 por cento e 80 por cento de sensibilidade, respectivamente, e 68 por cento, 85 por cento e 93 por cento de especificidade, respectivamente. No grupo de pacientes epilépticos, a positividade do ELISA foi 30 por cento, 51 por cento e 35 por cento com os antígenos Tso, Tcra-vf e Tcra-gp, respectivamente. Considerando o IB como teste confirmatório, a positividade foi de 16 por cento (14/88) no grupo total de pacientes epilépticos e 22 por cento (12/54) no grupo de pacientes epilépticos com TC positiva e sinais clínicos compatíveis com neurocisticercose. Foi encontrada correlação estatística significativa entre os resultados de ELISA ou IB e a fase da doença com quaisquer dos antígenos utilizados (p < 0,05). Os resultados indicam a necessidade de introduzir na rotina dos laboratórios o diagnóstico de neurocisticercose nos pacientes com convulsões epilépticas devido ao elevado risco de aquisição da cisticercose em nossa região e sua participação na etiologia da epilepsia.
Asunto(s)
Humanos , Animales , Anticuerpos Antihelmínticos/sangre , Antígenos Helmínticos , Epilepsia/parasitología , Neurocisticercosis/diagnóstico , Taenia solium/inmunología , Estudios de Casos y Controles , Ensayo de Inmunoadsorción Enzimática , Immunoblotting , Inmunoglobulina G , Neurocisticercosis/complicaciones , Sensibilidad y Especificidad , Tomografía Computarizada por Rayos XRESUMEN
Neurocysticercosis is the most frequent parasitic infection of the CNS and the main cause of acquired epilepsy worldwide. Seizures are the most common symptoms of the disease, together with headache, involuntary movements, psychosis and a global mental deterioration. Absolute diagnostic criteria include the identification of cysticerci, with scolex, in the brain by MRI imaging. We demonstrate here, for the first time, that T. solium DNA is present in the cerebrospinal fluid of patients. The PCR amplification of the parasite DNA in the CSF enabled the correct identification of 29/30 cases (96.7 %). The PCR diagnosis of parasite DNA in the CSF may be a strong support for the diagnosis of neurocysticercosis.
Asunto(s)
Antígenos Helmínticos/líquido cefalorraquídeo , ADN/líquido cefalorraquídeo , Neurocisticercosis , Taenia solium/genética , Animales , Humanos , Neurocisticercosis/líquido cefalorraquídeo , Neurocisticercosis/diagnóstico , Neurocisticercosis/microbiología , Reacción en Cadena de la Polimerasa/métodos , Sensibilidad y Especificidad , Taenia solium/inmunologíaRESUMEN
Sera from 88 patients from Santa Catarina and São Paulo states of Brazil, with epileptic seizures who underwent cerebral computed tomography (CT) were analyzed for the detection of antibodies to T. solium cysticercus by ELISA and Immunoblot (IB) with the following antigens: Taenia solium cysticercus total saline (Tso), Taenia crassiceps cysticercus vesicular fluid (Tcra-vf) and T. crassiceps cysticercus glycoproteins (Tcra-gp). ELISA carried out with Tso, Tcra-vf and Tcra-gp antigens showed 95%, 90% and 80% sensitivities, respectively, and 68%, 85% and 93% specificities, respectively. In the epileptic patients group, ELISA positivity was 30%, 51% and 35% with Tso, Tcra-vf and Tcra-gp antigens respectively. Considering the IB as the confirmatory test, the positivity was 16% (14/88) in the epileptic patients total group and 22% (12/54) in the epileptic patients with positive CT and signals of cysticercosis. We found a significant statistical correlation among ELISA or IB results and the phase of the disease when any antigens were used (p < 0.05). We emphasize the need to introduce in the laboratory routine the search for neurocysticercosis (NC) in patients presenting with epileptic seizures because of the high risk of acquiring NC in our region and its potential cause of epilepsy.
Asunto(s)
Anticuerpos Antihelmínticos/sangre , Antígenos Helmínticos , Epilepsia/parasitología , Neurocisticercosis/diagnóstico , Taenia solium/inmunología , Animales , Estudios de Casos y Controles , Ensayo de Inmunoadsorción Enzimática , Humanos , Neurocisticercosis/complicaciones , Sensibilidad y Especificidad , Tomografía Computarizada por Rayos XRESUMEN
The gold standard serodiagnostic assay for cysticercosis and neurocysticercosis, diseases caused by the metacestode of Taenia solium, uses lentil lectin-purified glycoprotein (LLGP) in a Western blot assay. We tested two antigens derived from LLGP, synthetic TS18var1 (sTS18var1) and recombinant GP50 antigen (rGP50), in an enzyme-linked immunosorbent assay (ELISA) using serum and cerebrospinal fluid (CSF) samples. The sensitivity for serum and CSF was 94.7% and 100% for rGP50 and 90.4% and 90.2% for sTS18var1, respectively. The specificity for serum and CSF samples was 93.8% and 100% for rGP50 and 90.3% and 98.0% for sTS18var1, respectively. The use of these antigens individually or combined as a diagnostic antigen cocktail eliminates the need for purification of antigens from parasite material and offers the advantage of using a simple and quantitative ELISA format.
Asunto(s)
Proteínas del Tejido Nervioso/sangre , Neurocisticercosis/diagnóstico , Taenia/inmunología , Teniasis/diagnóstico , Animales , Anticuerpos/sangre , Ensayo de Inmunoadsorción Enzimática/métodos , Humanos , Proteínas del Tejido Nervioso/genética , Neurocisticercosis/sangre , Neurocisticercosis/inmunología , Proteínas Recombinantes/sangre , Valores de Referencia , Sensibilidad y Especificidad , Pruebas Serológicas , Teniasis/sangre , Teniasis/inmunologíaRESUMEN
UNLABELLED: We analyzed cerebrospinal fluid (CSF) and blood serum from 55 patients with neurocysticercosis (NC) at different clinical stages. According to inflammatory activity in the CSF, three stages were identified: (1) reactive, when there was at least an increase in the number of cells; (2) weakly reactive, when significant alterations were found in the CSF, including an increase in gamma globulins, albeit without hypercytosis; (3) non-reactive, when there was neither hypercytosis nor increase in gamma globulins. Nineteen patients had the reactive form; 18 had the weakly reactive form; 18 displayed the non-reactive form. Local immunoproduction was intense in the reactive group, moderate in the weakly reactive group, and absent in the non-reactive group. The specific antibody index was raised in approximately 2/3 of patients with the reactive form, 2/3 in those with the weakly reactive form, and 1/3 in those with the non-reactive form. IN CONCLUSION: (1) the classical CSF syndrome in NC can present both in complete and partial modes; (2) local immunoproduction can occur in weakly reactive forms; (3) a raised specific antibody index can occur in the absence of an inflammatory reaction in the CSF.
Asunto(s)
Inmunoglobulina G/biosíntesis , Neurocisticercosis/inmunología , Adolescente , Adulto , Anciano , Femenino , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina G/líquido cefalorraquídeo , Leucocitosis/líquido cefalorraquídeo , Masculino , Persona de Mediana Edad , Neurocisticercosis/sangre , Neurocisticercosis/líquido cefalorraquídeo , Estudios Prospectivos , Síndrome , gammaglobulinas/líquido cefalorraquídeoRESUMEN
We report here the evaluation of an antigen from Taenia crassiceps cysticercus as a potential reagent in an enzyme-immunoelectrotransfer blotting assay (EITB) and an enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of neurocysticercosis (NC) using clinical specimens obtained from patients in different phases of the disease. Serum and cerebrospinal fluid (CSF) samples from 64 patients suspected of having NC according to clinical manifestation and brain computed tomography were tested by ELISA with Taenia solium total saline antigen (ELISA-Tso) and by immunoblotting with T. crassiceps glycoproteins antigen (EITB-gpTcra). Forty-five serum samples were also tested immunoblotting with T. solium glycoproteins antigen (EITB-gpTso) and 30 were tested by ELISA with T. crassiceps 14 kDa glycoprotein (ELISA-gp14Tcra). Serum samples from apparently healthy individuals without any parasitic disease and from patients with other parasitic diseases were included as controls. The results of ELISA-Tso analysis with CSF obtained from 64 patients with NC showed that 53 (83%) were reactive. EITB-gpTcra analysis with serum from the same group of patients showed a sensitivity of 91%. Results of EITB-gpTso and EITB-gpTcra analysis with serum samples demonstrated an agreement of 100% between both tests. ELISA-gp14Tcra was positive in 23 (77%) sera, 22 with paired CSF positive. When ELISA-gp14Tcra results were compared to EITB-Tso results, a relative sensitivity of 95% was observed. All serum samples from the control group were negative in ELISA-gp14Tcra and only one serum from an individual with Taenia saginata was reactive in this assay, showing a specificity of 99% for ELISA-gp14Tcra. This fraction was purified in only one step with a good yield for use in immunoassays. We suggest that the gp14Tcra antigen can be used for detecting anti-cysticercus antibodies in serum samples for epidemiological investigation purposes and also for diagnostic screening of NC patients.
Asunto(s)
Antígenos Helmínticos , Neurocisticercosis/diagnóstico , Taenia/inmunología , Animales , Anticuerpos Antihelmínticos/sangre , Brasil , Estudios de Casos y Controles , Ensayo de Inmunoadsorción Enzimática/métodos , Humanos , Neurocisticercosis/sangreRESUMEN
We analyzed cerebrospinal fluid (CSF) and blood serum from 55 patients with neurocysticercosis (NC) at different clinical stages. According to inflammatory activity in the CSF, three stages were identified: (1) reactive, when there was at least an increase in the number of cells; (2) weakly reactive, when significant alterations were found in the CSF, including an increase in gamma globulins, albeit without hypercytosis; (3) non-reactive, when there was neither hypercytosis nor increase in gamma globulins. Nineteen patients had the reactive form; 18 had the weakly reactive form; 18 displayed the non-reactive form. Local immunoproduction was intense in the reactive group, moderate in the weakly reactive group, and absent in the non-reactive group. The specific antibody index was raised in approximately 2/3 of patients with the reactive form, 2/3 in those with the weakly reactive form, and 1/3 in those with the non-reactive form. In conclusion: (1) the classical CSF syndrome in NC can present both in complete and partial modes; (2) local immunoproduction can occur in weakly reactive forms; (3) a raised specific antibody index can occur in the absence of an inflammatory reaction in the CSF
Asunto(s)
Humanos , Masculino , Femenino , Adolescente , Adulto , Persona de Mediana Edad , Inmunoglobulina G , Neurocisticercosis , gammaglobulinas , Inmunoglobulina G , Leucocitosis , Neurocisticercosis , Estudios Prospectivos , SíndromeRESUMEN
Considering the impact of cysticercosis on public health, especially the neurologic form of the disease, neurocysticercosis (NC), we studied the frequency of positivity of anti-Taenia solium cysticercus antibodies in serum samples from 1,863 inhabitants of Cássia dos Coqueiros, SP, a municipal district located 80 km from Ribeirão Preto, an area considered endemic for cysticercosis. The 1,863 samples were tested by enzyme linked immunosorbent assay (ELISA) using an antigenic extract from Taenia crassiceps vesicular fluid (Tcra). The reactive and inconclusive ELISA samples were tested by immunoblotting. Of the 459 samples submitted to immunoblotting, 40 were strongly immunoreactive to the immunodominant 18 and 14 kD peptides. Considering the use of immunoblotting as confirmatory due to its high specificity, the anti-cysticercus serum prevalence in this population was 2.1%.
Asunto(s)
Anticuerpos Antihelmínticos/sangre , Cisticercosis/sangre , Ensayo de Inmunoadsorción Enzimática , Immunoblotting , Taenia/inmunología , Adolescente , Adulto , Anciano , Animales , Antígenos Helmínticos/inmunología , Brasil , Estudios de Casos y Controles , Niño , Preescolar , Cysticercus/inmunología , Femenino , Humanos , Lactante , Recién Nacido , Masculino , Persona de Mediana Edad , Población Rural , Sensibilidad y Especificidad , Estudios SeroepidemiológicosRESUMEN
Considering the impact of cysticercosis on public health, especially the neurologic form of the disease, neurocysticercosis (NC), we studied the frequency of positivity of anti-Taenia solium cysticercus antibodies in serum samples from 1,863 inhabitants of Cássia dos Coqueiros, SP, a municipal district located 80 km from Ribeiräo Preto, an area considered endemic for cysticercosis. The 1,863 samples were tested by enzyme linked immunosorbent assay (ELISA) using an antigenic extract from Taenia crassiceps vesicular fluid (Tcra). The reactive and inconclusive ELISA samples were tested by immunoblotting. Of the 459 samples submitted to immunoblotting, 40 were strongly immunoreactive to the immunodominant 18 and 14 kD peptides. Considering the use of immunoblotting as confirmatory due to its high specificity, the anti-cysticercus serum prevalence in this population was 2.1 percent
Asunto(s)
Humanos , Animales , Masculino , Femenino , Recién Nacido , Lactante , Preescolar , Niño , Adolescente , Adulto , Persona de Mediana Edad , Anticuerpos Antihelmínticos , Antígenos Helmínticos , Cisticercosis , Inmunoensayo , Taenia , Brasil , Estudios de Casos y Controles , Cysticercus , Ensayo de Inmunoadsorción Enzimática , Immunoblotting , Población Rural , Sensibilidad y Especificidad , Estudios SeroepidemiológicosRESUMEN
We describe the production of the potential monoclonal antibodies (MoAbs) using BALB/c mice immunized with vesicular fluid (VF)-Tcra (T. crassiceps) antigen. Immune sera presented anti-VF-Tcra (<20kD) IgG and IgM antibodies with cross-reactivity with T. solium (Tso) antigen (8-12, 14, and 18 kD). After cell fusion, we selected 33 anti-Tcra and anti-Tso reactive IgM-clones and 53 anti-Tcra specific IgG-clones, 5 of them also recognizing Tso antigens. Two clones identified the 8-14 and 18kD peptides of VF-Tcra.
Asunto(s)
Animales , Femenino , Ratones , Anticuerpos Antihelmínticos/biosíntesis , Anticuerpos Monoclonales/biosíntesis , Antígenos Helmínticos/inmunología , Taenia/inmunología , Reacciones Cruzadas , Cisticercosis/inmunología , Immunoblotting , Inmunoglobulina G/inmunología , Inmunoglobulina G/aislamiento & purificación , Inmunoglobulina M/inmunología , Inmunoglobulina M/aislamiento & purificación , Ratones Endogámicos BALB CRESUMEN
Foram analisados os resultados dos testes imnoenzimáticos ELISA (Roche Diagnóstica) para anticorpos anti-HAV das classes IgG (HAV-G) e IgM (HAV-M) obtidos no período de dezembro de 1995 a novembro de 1996. Os 1177 pacientes estavam assim distribuídos: 1042 (88,5 por ciento) adultos (= 15 anos), sendo 49,4 por ciento do sexo masculino e 50,6 por ciento feminino, e 135 crianþas (54,1 por ciento masculino e 45,9 feminino). Foram obtidos os seguintes índices de positividade para HAV-G: 76,2 por ciento para adultos (77,5 por ciento dos homens e 45,9 por ciento das mulheres) e 48,9 por ciento para as crianþas (56,2 por ciento entre os meninos e 40,3 por ciento entre as meninas) Os adultos apresentaram maior freq³Ûncia de positividade que as crianþas (p<0,05). Os contrário foi observado para anticorpos IgM: 4,6 por ciento de positividade para adultos e 29,6 por ciento entre as crianþas (AU)
Asunto(s)
Humanos , Masculino , Femenino , Preescolar , Adolescente , Adulto , Anciano , Hepatitis A/diagnóstico , Factores de Edad , Estaciones del Año , Ensayo de Inmunoadsorción Enzimática , Hepatitis A/transmisión , Inmunoglobulina G/análisis , Inmunoglobulina G/diagnósticoRESUMEN
Foram analisados os resultados dos testes imnoenzimáticos ELISA (Roche Diagnóstica) para anticorpos anti-HAV das classes IgG (HAV-G) e IgM (HAV-M) obtidos no período de dezembro de 1995 a novembro de 1996. Os 1177 pacientes estavam assim distribuídos: 1042 (88,5 por ciento) adultos (= 15 anos), sendo 49,4 por ciento do sexo masculino e 50,6 por ciento feminino, e 135 crianças (54,1 por ciento masculino e 45,9 feminino). Foram obtidos os seguintes índices de positividade para HAV-G: 76,2 por ciento para adultos (77,5 por ciento dos homens e 45,9 por ciento das mulheres) e 48,9 por ciento para as crianças (56,2 por ciento entre os meninos e 40,3 por ciento entre as meninas) Os adultos apresentaram maior freqüência de positividade que as crianças (p<0,05). Os contrário foi observado para anticorpos IgM: 4,6 por ciento de positividade para adultos e 29,6 por ciento entre as crianças
Asunto(s)
Humanos , Masculino , Femenino , Preescolar , Adolescente , Adulto , Anciano , Factores de Edad , Hepatitis A/diagnóstico , Ensayo de Inmunoadsorción Enzimática , Hepatitis A/transmisión , Inmunoglobulina G , Inmunoglobulina G/análisisRESUMEN
A concentração de ferritina sérica está diretamente relacionada ao estoque de ferro no organismo. Esta determinação é útil no diagnóstico e tratamento da deficiência de ferro, e em estados onde ocorre sobrecarga de ferro tais como na talassemia e anemia sideroblástica. Os testes utilizados atualmente são baseados em métodos imunológicos, empregando anticorpos anti-ferritina. Os testes utilizados atualmente são baseados em métodos imunológicos, empregando anticorpos anti-ferritina. Alguns fabricantes de kits recomendam o uso somente de soro, e outros de soro e plasma para esta determinação. Para a obtenção de plasma usam-se anticoagulantes; dos disponíveis, o EDTA é o anticoagulante de escolha no Laboratório de Hematologia. O objetivo deste trabalho foi determinar a ferritina sérica e plasmática (com uso EDTA) de 109 amostras através do kit IMxR Abbott (enzimaimunoensaio com micropartículas). Com o uso de anticoagulante K3EDTA houve uma grande variação nos resultados obtidos, mostrando que os valores da ferritina plasmática apresentaram-se muito mais baixos (em algumas amostras esta redução foi de até 100%) quando comparados com os valores obtidos no soro do mesmo paciente. Em média, houve uma redução significativa em torno de 61% dos valores da ferritina obtida no respectivo soro, confirmando que o EDTA não deve ser utilizado como anticoagulante na determinação da ferritina.
Asunto(s)
Humanos , Femenino , Deficiencias de Hierro/diagnóstico , Hierro , Estado NutricionalRESUMEN
A concentração de ferritina sérica está diretamente relacionada às reservas de ferro do organismo. A proposta de nosso trabalho foi comparar os resultados obtidos na determinação da ferritina sérica por dois métodos imunológicos automatizados, uma vez que os resultados obtidos podem implicar ou não em condutas terapêuticas. Foram analisadas 206 amostras de soro por nefelometria usando o kit N-Látex-Ferritin da Behring e por enzimaimunoensaio em micropartículas usando o kit IMx Abbott. O coeficiente de correlação entre os dois métodos foi de r² = 0,94. No entanto, quando o método de enzimaimunoensaio foi utilizado, a média dos resultados obtidos foi significativamente mais altos que a média dos resultados do método de nefelometria (p < 0,001). A seguinte equação de regressão foi obtida: y = 0,83x + 1,172, onde y é o resultado da nefelometria e x é o resultado do método IMx Abbott. A despeito da alta correlação obtida, os resultados confirmam a importância de se especificar a metodologia aplicada nos resultados reportados
Asunto(s)
Humanos , Técnicas de Laboratorio Clínico , Ferritinas/análisis , Informática Médica , Métodos de Análisis de Laboratorio y de CampoRESUMEN
Foi padronizado o teste de hemaglutinação (HA) empregando hemácias de ganso formolizadas, taninizadas e sensibilizadas com extrato antigênico de líquido vesicular de C. longicollis (HA-CI) e extrato salino total de C. cellulosae (HA-Cc). Foram ensaiados 61LCR de dois grupos: 41 de pacientes com neurocisticercose e 20 de grupo controle, respectivamente, reativos e não-reativos no teste ELISA empregando C. cellulosae. Nos LCR do grupo controle não foi observada reatividade e 34 (82,9%) e 35 (85,4%) LCR de doentes foram reativos, respectivamente, nos testes HA-CI e HA-Cc. O estudo da estabilidade dos reagentes pronto para uso mostrou vantagens para o armazenamento a 4°C, em glicerol a 50%, por até 6 meses. Os resultados obtidos indicam que o reagente utilizando Cysticercus longicollis e estabilizado com glicerol pode ser empregado como alternativa no diagnóstico imunológico da neutocisticercose