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1.
Rev. Inst. Med. Trop. Säo Paulo ; 49(6): 379-384, Nov.-Dec. 2007. ilus, tab
Article in English | LILACS | ID: lil-470521

ABSTRACT

The Western blot technique was used to demonstrate the presence of antibodies in the blood of dogs that presented canine visceral leishmaniasis. This technique was used against some specific molecules present in the lysate of the promastigote form of Leshmania chagasi.Through the association of the results of the Western blot technique with the morphological alterations seen as a result of the serum neutralization technique performed in McCoy cells (which mimetizes the macrophage) it was possible to observe the role of some molecules of great relevance in determining the disease in symptomatic dogs as well as that of some other molecules associated with asymptomatic infected dogs that may become transmitters as well as differentiating them as asymptomatic resistant dogs. In the sera analyses carried out during the immunobloting a variation of 9 to 27 immunoreacting bands was observed, which were then compared using Dice's similarity coefficient. In the dendrogram constructed on the basis of the coefficient, 50 percent similarity was observed among the total number of reagent bands with the promastigote lysate, thus creating five groups. The main difference observed related to the clinical condition of the dogs: symptomatic and asymptomatic dogs were found in separate groups. The asymptomatic group of dogs was distributed in two different places in the dendrogram because they presented two different behavior patterns regarding the cellular morphology in the serum neutralization reaction: the presence or absence of cellular lysis. According to this analysis it is possible to evaluate the immune status and associate it with specific markers observed in the reaction found in the Western blot strips.


A técnica de Western blot foi utilizada para demonstrar a presença de anticorpos do soro de cães, que apresentavam leishmaniose visceral canina, contra algumas moléculas específicas no lisado da forma promastigota de Leshmania chagasi.Através da associação da técnica de Western blot com as alterações morfológicas observadas como resultado da técnica de soro-neutralização em células McCoy (que mimetizam o macrófago) foi possível observar o papel de algumas moléculas de maior relevância para a determinação da doença em cães sintomáticos bem como o papel de outras moléculas na predição de cães infectados assintomáticos com o potencial de serem transmissores e ainda diferenciá-los como cães assintomáticos resistentes. Na análise dos soros durante a reação de immunoblotting observou-se uma variação de 9 a 27 bandas imunorreagentes, que foram comparadas utilizando-se o coeficiente de similaridade de Dice. No dendrograma construído com base no coeficiente, observou-se 50 por cento de similaridade entre as bandas totais reagentes com o lisado de promastigota formando cinco agrupamentos. A principal diferença foi observada com respeito à condição clínica, ou seja, cães sintomáticos e assintomáticos ficaram em grupos separados. Os soros dos cães assintomáticos distribuídos em dois grupos diferentes do dendrograma apresentaram padrões de comportamento diferentes, quanto à morfologia celular na reação de soro-neutralização, ou seja, a presença ou ausência de lise celular. De acordo com esta análise foi possível avaliar o status imunitário e associá-lo com determinados marcadores específicos observados na reação encontrada nas fitas de Western blot.


Subject(s)
Animals , Dogs , Dog Diseases/immunology , Leishmania infantum/immunology , Leishmaniasis, Visceral/veterinary , Blotting, Western , Dog Diseases/diagnosis , Enzyme-Linked Immunosorbent Assay/veterinary , Leishmania infantum/genetics , Leishmaniasis, Visceral/diagnosis , Leishmaniasis, Visceral/immunology , Neutralization Tests , Polymerase Chain Reaction
2.
Rev. Inst. Med. Trop. Säo Paulo ; 48(6): 337-341, nov.-dez. 2006. ilus
Article in English | LILACS | ID: lil-439866

ABSTRACT

The kinetics of growth of Leishmania performed in vitro after internalization of the promastigote form in the cell and the occurrence of the transformation of the parasite into the amastigote form have been described by several authors. They used explants of macrophages in hamster spleen cell culture or in a human macrophage lineage cell, the U937. Using microscopy, the description of morphologic inter-relationship and the analysis of the production of specific molecules, it has been possible to define some of the peculiarities of the biology of the parasite. The present study shows the growth cycle of Leishmania chagasi during the observation of kinetic analysis undertaken with a McCoy cell lineage that lasted for a period of 144 hours. During the process, the morphologic transformation was revealed by indirect immunofluorescence (IF) and the molecules liberated in the extra cellular medium were observed by SDS-PAGE at 24-hour intervals during the whole 144-hour period. It was observed that in the first 72 hours the promastigote form of L. chagasi adhered to the cell membranes and assumed a rounded (amastigote-like) form. At 96 hours the infected cells showed morphologic alterations; at 120 hours the cells had liberated soluble fluorescent antigens into the extra cellular medium. At 144 hours, new elongated forms of the parasites, similar to promastigotes, were observed. In the SDS-PAGE, specific molecular weight proteins were observed at each point of the kinetic analysis showing that the McCoy cell imitates the macrophage and may be considered a useful model for the study of the infection of the Leishmania/cell binomial.


Cinéticas de crescimento de Leishmania realizadas in vitro após a internalização da forma promastigota na célula e a ocorrência da transformação do parasito na forma amastigota foram descritas por vários autores, seja com a utilização de explantes de macrófagos em células de baço de hamster ou atualmente da célula de linhagem de macrófago humano U937. Aliando a microscopia à descrição das inter-relações morfológicas e à síntese de moléculas específicas foi possível esclarecer pontos sobre a biologia do parasito. O presente estudo mostra o acompanhamento do ciclo de crescimento da Leishmania chagasi em uma cinética realizada com células de linhagem McCoy, no período de 144 horas. Durante o processo, as transformações morfológicas foram reveladas pela reação de imunofluorescência indireta (RIFI) e as moléculas liberadas no meio extracelular foram observadas pelo método de SDS-PAGE, em intervalos de 24 horas no período de 144 horas. Observou-se que nas primeiras 72 horas, a forma promastigota da L. chagasi fica aderida à membrana das células com aspecto arredondado (amastigota-like). Em 96 horas as células infectadas apresentaram alterações morfológicas; em 120 horas, as células liberaram, para o meio extracelular, antígenos fluorescentes solúveis; e em 144 horas foram observadas novas formas alongadas dos parasitos como se fossem promastigotas. No SDS-PAGE, proteínas com pesos moleculares específicos são observadas em cada ponto da cinética, mostrando que a célula McCoy parece mimetizar o macrófago e que pode ser um modelo útil para o estudo da infecção do binômio leishmânia/célula.


Subject(s)
Animals , Cricetinae , Leishmania infantum/growth & development , Culture Media, Conditioned , Cell Line/parasitology , Electrophoresis, Polyacrylamide Gel , Fluorescent Antibody Technique, Indirect , Kinetics , Time Factors
3.
Rev. Inst. Med. Trop. Säo Paulo ; 45(4): 205-211, July-Aug. 2003. ilus, tab
Article in English | LILACS | ID: lil-345384

ABSTRACT

Several studies have recently shown the use of recombinant rabies virus as potential vector-viral vaccine for HIV-1. The sequence homology between gp 120 and rabies virus glycoprotein has been reported. The McCoy cell line has therefore been used to show CD4+ or CD4+ like receptors. Samples of HIV-1 were isolated, when plasma of HIV-1 positive patients was inoculated in the McCoy cell line. The virus infection was then studied during successive virus passages. The proteins released in the extra cellular medium were checked for protein activity, by exposure to SDS Electrophoresis and blotting to nitro-cellulose filter, then reacting with sera of HIV positive and negative patients. Successive passages were performed, and showed viral replication, membrane permeabilization, the syncytium formation, and the cellular lysis (cytopathic effect). Flow cytometry analysis shows clear evidence that CD4+ receptors are present in this cell line, which enhances the likelihood of easy isolation and replication of HIV. The results observed allow the use of this cell line as a possible model for isolating HIV, as well as for carrying out studies of the dynamics of viral infection in several situations, including exposure to drugs in pharmacological studies, and possibly studies and analyses of the immune response in vaccine therapies


Subject(s)
Humans , HIV-1 , Virus Replication , CD4 Antigens , Blotting, Western , CD4-Positive T-Lymphocytes , Cell Line , Flow Cytometry , Fluorescent Antibody Technique, Indirect , Virus Replication
4.
Rev. Inst. Med. Trop. Säo Paulo ; 44(1): 7-12, Jan.-Feb. 2002. ilus, tab, graf
Article in English | LILACS | ID: lil-307235

ABSTRACT

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


Subject(s)
Humans , Animals , Male , Female , Infant, Newborn , Infant , Child, Preschool , Child , Adolescent , Adult , Middle Aged , Antibodies, Helminth , Antigens, Helminth , Cysticercosis , Immunoassay , Taenia , Brazil , Case-Control Studies , Cysticercus , Enzyme-Linked Immunosorbent Assay , Immunoblotting , Rural Population , Sensitivity and Specificity , Seroepidemiologic Studies
5.
Bol. chil. parasitol ; 56(1/2): 36-42, ene.-jun. 2001. tab
Article in English | LILACS | ID: lil-300167

ABSTRACT

Se utilizó la técnica del inmunoblot para el diagnóstico de la cisticercosis porcina usando un antígeno total de cisticercos de taenia crassiceps. Fueron analizados 13 sueros del cerdo con cisticercosis, 30 sueros controles negativos y ocho sueron del cerdo con hidatidosis, así como nueve del suino con macracantorincosis, 10 con ascaridiosis y ocho con pulmonía. El uso de este antígeno en el inmunoblot con suero de cerdos no se había publicado previamente. El inmunoblot fue padronizado por análisis de titulación en bloque mostrando 100,0 por ciento de sensibilidad y 96,7 por ciento de especificidad. Los péptidos específicos para la cisticercosis en orden de frecuencia fueron: 72-68 kD (100 por ciento), 16-15 kD (77 por ciento), 39-36 kD (62 por ciento), 18-17 kD (54 por ciento, 21 kD (31 por ciento), 14 kD (23 por ciento), 25-23 kD (8 por ciento), y 20-19 kD (8 por ciento). Reacción cruzada (72-68 y 18-17 kD) sólo se descubrió en una muestra (12,5 por ciento) de cerdo con hidatidosis. Debido a sus altas tasas de desempeño, el inmunoblot debe ser útil para confirmar el diagnóstico de cisticercosis porcina y es más eficaz que otras pruebas empleadas para este propósito, como examen de la lengua, examen anatomopatológico y ELISA


Subject(s)
Animals , Cysticercosis , Immunoblotting , Swine Diseases , Taenia , Antigens, Helminth , Cysticercosis , Sensitivity and Specificity , Swine , Swine Diseases , Taenia
6.
Rev. Inst. Med. Trop. Säo Paulo ; 42(3): 175-7, May-Jun. 2000. ilus
Article in English | LILACS | ID: lil-262699

ABSTRACT

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.


Subject(s)
Animals , Female , Mice , Antibodies, Helminth/biosynthesis , Antibodies, Monoclonal/biosynthesis , Antigens, Helminth/immunology , Taenia/immunology , Cross Reactions , Cysticercosis/immunology , Immunoblotting , Immunoglobulin G/immunology , Immunoglobulin G/isolation & purification , Immunoglobulin M/immunology , Immunoglobulin M/isolation & purification , Mice, Inbred BALB C
7.
Rev. Inst. Med. Trop. Säo Paulo ; 42(2): 71-9, Mar.-Apr. 2000. ilus, tab, graf
Article in English | LILACS | ID: lil-256388

ABSTRACT

In the present study ELISA was standardized for the diagnosis of swine cysticercosis based on necropsy parameters and confirmed positive and negative control sera. Serum samples from pigs with other infections were also assayed to determine possible cross-reactions. Four antigens were assayed: from Taenia crassiceps vesicular fluid (VF-Tcra) and crude larvae extract (T-Tcra), and from Taenia solium extracts of scolex (S-Ts) and of larvae (T-Ts). A checkerboard evaluation of antigen, serum and conjugate dilutions, as well as the use of Tween-20 and skim cow milk in wash and blocking solution had a marked effect on improving ELISA performance. All the antigens showed a good performance, but VF-Tcra was the best, with 96.0 per cent and 80.0 per cent sensitivities for cut-offs respectively at 2sd and 3sd, and corresponding specificities of 97.5 per cent and 100.0 per cent. Cross-reactivity was observed only with hydatidosis and ascaridiosis. In view of the high performance observed, the ELISA test should be recommended for the diagnosis of cysticercosis in suspected swine in slaughterhouses and for the screening of cysticercosis in swine production. These results will support integrated measures of cysticercosis control throughout the chain of swine production, effectively contributing to public health.


Subject(s)
Animals , Antigens, Helminth/isolation & purification , Cysticercosis/veterinary , Cysticercus/immunology , Swine Diseases/diagnosis , Antigens, Helminth/blood , Biomarkers/blood , Cysticercosis/diagnosis , Enzyme-Linked Immunosorbent Assay , Swine
8.
Mem. Inst. Oswaldo Cruz ; 93(6): 727-32, Nov.-Dec. 1998. tab
Article in English | LILACS | ID: lil-223871

ABSTRACT

Seropositivity for Chagas disease was evaluated in 834 children aged between 7 and 14 from the Municipal Teaching System in the District of Londrina, State of Paraná. A seroprevalence rate of 0.1 per cent was found through the use of an indirect immunofluorescent test and an enzyne-linked immunosorbent assay. This low rate of seroprevalence provides evidence that the vectorial transmission of Chagas disease has been eliminated in Londrina. The main reason for the elimination of vectorial transmission of Trypanosoma cruzi infection, as evaluated by serological tests, may be a remarkable change in the economic structure of the northern region of Paraná in the 1960's. At that time coffee production was almost completely replaced by soy beans, wheat and grazing in the rural areas. This change deeply affected the rural ecology and caused an exodus of the population from rural to urban areas as well as a decrease in the total number of the population of that region. The measures introduced for controlling the disease through the Program of Chagas Disease Control established by the Fundaçäo Nacional de Saúde of the Brazilian Ministry of Health, certainly, had a positive impact on the reduction of American trypanosomiasis prevalence in the area under study. However, it does not seem that this was the most relevant factor responsable for the elimination of vectorial transmission of Chagas disease in Londrina.


Subject(s)
Humans , Child , Chagas Disease/epidemiology , Trypanosoma cruzi/parasitology , Brazil/epidemiology , Communicable Disease Control
9.
Mem. Inst. Oswaldo Cruz ; 93(supl.1): 279-82, Oct. 1998. tab
Article in English | LILACS | ID: lil-218689

ABSTRACT

The high sensitivity and the possibility of automation of the enzyme-linked-immunosorbent-assay (ELISA) has indicated this technique as one of the most useful serological test for epidemiological studies. In the present study, an ELISA for detection of IgG antibodies against adult worm antigens (IgG-ELISA) was investigated for epidemiological purposes, in a rural area of the municipality of Itariri (Säo Paulo, Brazil). Blood on filter paper (1,180 samples) from about 650 school children were submitted to ELISA and the data compared to the results of the parasitogical method of Kato-Katz and also to the IgM-IFT (immunofluorescence test for IgM antibodies to gut associated antigens). The prevalence rates respectively of 8.5 per cent, 43.0 per cent and 56.2 per cent by the Kato-Katz, IgG-ELISA, and IgM-IFT methods suggest the poor sensitivity of the parasitological method for detection of Schistosoma mansoni eggs in individuals with low worm burden, situation commonly observed in low endemic areas. These results can partially explain the poor degree of agreement between the IgG-ELISA and the Kato-Katz, as suggested by the Kappa index of 0.170. Otherwise, the Kappa index of 0.675 showed substantial agreement between the two serological tests. Some discrepancy of results between the two serological techniques must be better investigated.


Subject(s)
Humans , Child , Brazil , Enzyme-Linked Immunosorbent Assay , Immunoglobulin G , Schistosomiasis , Schistosoma mansoni/parasitology
10.
Rev. bras. anal. clin ; 30(2): 58-60, 1998. graf
Article in Portuguese | LILACS | ID: lil-525182

ABSTRACT

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


Subject(s)
Cerebrospinal Fluid , Clinical Laboratory Techniques , Hemagglutination , Neurocysticercosis/diagnosis
11.
Hansen. int ; 23(1/2): 35-48, 1998. tab, graf
Article in Portuguese | LILACS | ID: lil-426898

ABSTRACT

Testes sorologicos de hanseniase utilizando o antigeno glicolipideo fenolico-1 (PGL-1) abriram varias possibilidades para o estudo do comportamento epidemiologico dessa doença. O objetivo do trabalho foi avaliar os resultados da aplicaçao do teste de Elisa anti-PGL-1 em uma comunidade urbana com alta endemicidade de hanseniase no estado de Sao Paulo. Nela se conseguiu, na epoca do estudo, coeficientes de detecçao e prevalencia de 27,1 e 167,2 casos por 10.000 habitantes, respectivamente. Foram recenseados 8.491 residentes na area urbana e destes 6.666 foram avaliados com o teste Elisa IgM anti-PGL-1. A sorologia se revelou positiva em 9,0 por cento da populaçao geral, sendo que, para as mulheres, a taxa de positividade encontrada foi de 10,1 por cento e para os homens de 7,6 por cento...


Subject(s)
Leprosy/diagnosis , Leprosy/epidemiology , Serology
12.
Hansen. int ; 22(2): 35-43, jul.-dez. 1997. tab, graf
Article in Portuguese | LILACS, SES-SP | ID: lil-222031

ABSTRACT

Os testes sorológicos para diagnóstico de hanseníase, usando o glicolipídeo-fenólico, considerado antígeno específico do M. leprae têm aberto algumas possibilidades de estudo do comportamento epidemiológico desta doença. Foi realisado um estudo para medir a sensibilidade e especificidade de um teste de ELISA anti PGL-1, usando material e técnica proveniente de Cuba (UMELISA). Foram testados 84 doentes e 112 controles sadios. A sensibilidade do teste foi maior para o grupo de doentes multibacilares, sendo mais alto entre os casos classificados como virchovianos (V), seguido dos dimorfos (D). No grupo de multibacilares, considerando o limiar de reatividade de 0,200, observou-se que apenas 75,0(por cento) de pacientes V e 50,0(por cento) de D foram positivos ao teste e, no limiar de 0,300, apenas 64,3(por cento) dos V e 40,0(por cento) dos D ainda mostravam positividade. Os doentes indeterminados apresentaram maior proporçäo de soropositivos do que os tuberculóides e isto talvez traduza a polarizaçäo de alguns casos para formas multibacilares. A especificidade do teste foi de 87,5(por cento) no limiar de 0,200 e de 99,1(por cento) no de 0,300. Este teste, à semelhança de outros, apresentou alta especificidade e baixa sensibilidade, resultando em grande percentual de falso-negativos. Para uma doença em que os testes sorológicos säo escassos, apesar de näo se recomendar o uso indiscriminado do teste para triagem de casos na populaçäo geral, a comunidade de seu aprimoramento tecnológico deve ser estimulada. Conseguir-se-ia, assim, avançar na pesquisa de instrumentos cada vez mais sensíveis e específicos para melhorar o diagnóstico precoce e, desse modo intervir mais oportunamente na cadeia de transmissäo


Subject(s)
Enzyme-Linked Immunosorbent Assay , Glycolipids/adverse effects , Leprosy/diagnosis , Microbial Sensitivity Tests , Epidemiologic Studies , Serologic Tests
13.
Arq. neuropsiquiatr ; 55(1): 8-11, mar. 1997. ilus
Article in English | LILACS | ID: lil-194696

ABSTRACT

The detection of IgE is technically difficult because of its reduced concentrations in serum, and even lower concentrations in cerebrospinal fluid (CSF). In the present investigation we studied 86 CSF samples using animmunoenzymatic method with an anti-IgE-alkaline phosphatase conjugate and a fluorigenic substrate. The samples were from three groups: A) 29 patients with neurocysticercosis (NC), B) 36 patients with different neurologic disorders (neurosyphilis, neurotuberculosis, meningitis, tumors, hemorrhage) and C) 21 discharged individuals who had been hospitalized for bacterial meningitis. The results obtained were: A) 0.05 to 3.00 IU/ml (0.76 + 0.79), B) 0.00 to 1.50 IU/ml (0.23 + 0.34) and C) 0.05 to 1.25 IU/ml (0.34 + 0.34). The present results suggest that IgE appears to play a role in the pathogeny of NC and that efforts should be made to standardize a test for the detection of specific IgE antibodies.


Subject(s)
Humans , Central Nervous System Diseases/cerebrospinal fluid , Central Nervous System Diseases/immunology , Cysticercosis/cerebrospinal fluid , Cysticercosis/immunology , Immunoglobulin E/cerebrospinal fluid , Immunoenzyme Techniques
14.
Rev. Inst. Med. Trop. Säo Paulo ; 39(1): 29-33, jan.-fev. 1997. tab, ilus
Article in English | LILACS | ID: lil-195546

ABSTRACT

Foi padronizado o teste de hemaglutinacao (HA) utilizando as hemacias formolizadas e taninizadas de ganso sensibilizadas com extrato salino total de C. cellulosae (HA-Cc) e liquido vesicular de Cysticercus longicollis (HA-Cl). Foram ensaiadas 61 amostras de liquido cefalorraquiano (LCR), 41 de pacientes com neurocisticercose e 20 de um grupo de controle, respectivamente, regentes e nao-regentes no teste ELISA utilizando antigenos de C. cellulosae...


Subject(s)
Animals , Female , Mice , Antigens, Heterophile , Cysticercosis/diagnosis , Neurologic Manifestations , Antigens, Heterophile/immunology , Cysticercosis/cerebrospinal fluid , Cysticercosis/immunology , Enzyme-Linked Immunosorbent Assay , Indicators and Reagents , Hemagglutination Tests/methods
15.
Rev. Inst. Med. Trop. Säo Paulo ; 38(6): 391-6, nov.-dez. 1996. ilus, tab
Article in English | LILACS | ID: lil-186860

ABSTRACT

Foi desenvolvido o teste dot-Elisa para deteccao de anticorpos em liquido cefalorraquiano (LCR) no diagnostico imunologico da neurocisticercose humana, utilizando antigenos de membrana e escolex de Cysticercus cellulose (M+E-Cc) e, alternativamente, membrana (M) e liquido vesicular (LV) de Cysticercus longicollis (Cl) covalentemente ligados a um novo suporte constituido de tecido de poliester-resina de N-metilolacrilamida (dot-TR). O teste foi realizado a temperatura ambiente, com tempos de incubacao reduzidos e sem necessidade de cuidados na manipulacao do suporte...


Subject(s)
Humans , Antibodies, Helminth/immunology , Cysticercus/immunology , Enzyme-Linked Immunosorbent Assay , Cysticercosis/cerebrospinal fluid , Neurologic Manifestations , Resins/classification
16.
Rev. Inst. Med. Trop. Säo Paulo ; 38(4): 259-63, jul.-ago. 1996. ilus, tab
Article in English | LILACS | ID: lil-182827

ABSTRACT

A cepa ORF de Cysticercus longicollis (Cl) representa importante modelo para estudo de antigenos heterologos no imunodiagnostico da neurocisticercose (NC). Foi padronizada a tecnica de imunoperoxidase (IP) empregando suspensao antigenica particulada. Amostras de liquido cefalorraquiano (LCR) foram incubadas sobre o antigeno fixado em laminas de microscopia, o conjugado empregado foi anti-IgG-Peroxidase, a reacao enzimatica iniciou-se ao cobrirem-se as laminas com solucao cromogena (Diaminobenzidina/H2O2). Apos lavagens em agua destilada, a lamina foi corada com verde malaquita a 2 por cento em agua. De 21 LCR de pacientes com NC, 19 (90,5 por cento) foram reativos e 8 (100 por cento) LCR do grupo controle foram nao reativos. Os resultados do teste IP-Cl ensaiando 127 LCR de pacientes com suspeita de NC mostrou 89,7 por cento de concordancia com o teste ELISA empregando extrato salino de Cysticercus cellulosae (Cc) e 94,2 por cento de concordancia com o teste IP-Cc


Subject(s)
Animals , Female , Rats , Cysticercosis/cerebrospinal fluid , Immunoenzyme Techniques , Cysticercosis/immunology , Cysticercosis/parasitology , Cysticercus/classification , Cysticercus/ultrastructure , Microscopy , Neurologic Manifestations
17.
Rev. Soc. Bras. Med. Trop ; 29(2): 137-44, Mar.-Apr. 1996. ilus, graf, tab
Article in Portuguese | LILACS, SES-SP | ID: lil-187140

ABSTRACT

A new reagent was designed to the indirect hemagglutination test (IHATIAL), utilizing goose red blood cells as inert matrix and standardized for the field diagnosis of American trypanosomiasis. The objective was to substitute the lyophilized or frozen reagent of IHAT produced routinely using human erythrocytes in the Adolfo Lutz Institute (Säo Paulo/Brazil). The standardized reagent presented a long stability in liquid suspension, and was evaluated in 137 serum samples from patient with and without Chagas' disease, by IHATIAL. The diagnostic performance of this test was similar to the IHAT utilizing human erythrocytes and to that of a commercial IHAT kit. The sensitivity was 1.00, specificity 0.98, predictive value of positive 0.96 and of negative 1.00. Different batches of reagent successively produced proved to be reproducible in a quality control method. The new reagent is more economic than the former reagent, it can be produced easily and may be applicable to the seroepidemiologic studies.


Subject(s)
Humans , Animals , Geese/immunology , Chagas Disease/diagnosis , Hemagglutination Tests/methods , Sensitivity and Specificity , Geese/blood , Indicators and Reagents
18.
Rev. Inst. Med. Trop. Säo Paulo ; 36(5): 475-8, set.-out. 1994. ilus, tab
Article in English | LILACS | ID: lil-154324

ABSTRACT

Com a finalidade de melhorar o diagnostico da leptospirose humana, padronizou-se o teste dot-ELISA para a pesquisa de anticorpos especificos da classe IgM na saliva. Empregaram-se amostras de saliva e soro coletadas simultaneamente de 20 pacientes com a forma icterohemorragica da doenca, de 10 pacientes com outras patologias e 5 controles negativos. Culturas de Leptospira em meio EMJH, dos sorovares: icterohaemorrhagiae, canicola, hebdomadis, brasiliensis e cynopteri, foram utilizadas como antigeno, na concentracao proteica individual de 0,2 ug/ul, misturadas em volumes iguais. Na fase solida do teste empregou-se tecido de poliester impregnado com resina de N-metilol-acrilamida....


Subject(s)
Immunoglobulin M/immunology , Leptospirosis/diagnosis , Saliva/microbiology , Immunoenzyme Techniques , Leptospirosis/blood , Resins, Synthetic/classification
19.
CCS ; 13(1): 39-46, jan.-mar. 1994. tab, graf
Article in Portuguese | LILACS | ID: lil-190841

ABSTRACT

O Peso ao Nascer(PN) segundo vários autores é um importante indicador para avaliar o estado de saúde do recém-nascido (RN). Objetivou-se fazer um estudo prospectivo do RN. Foram estudados 2.270 RN's nas maternidades do município de joäo Pessoa/PB a partir de dados de arquivos, com distribuiçäo de peso: peso adequado (PA) 68,2 por cento (15.744), peso insuficiente (PI) 23,2 por cento (536), baixo peso ao nascer (BPN) 8,6 por cento (198). Em relaçäo ao sexo, os meninos apresentaram 72,7 por cento (892) de PA e 7,4 por cento (90) de BPN, e nas meninas 63,1 por cento (692) de PA 9,8 por cento (108) de BPN. A estrutura em comparaçäo ao padräo (Alcântara/Marcondes), 50,6 por cento dos meninos e 51,6 por cento das meninas revelaram déficits estaturais . Houve um incrementos de 2,3 por cento no BPN em relaçäo a 1989 (6,3 por cento). Quanto ao sexo, estatura, e número de conceptos näo houve alteraçöes significativas . Seriam os fatores sócio-econômicos responsáveis pelo incremento ou uma melhoria dos serviços na aferiçäo e registro da antropometria dos recém-nascidos?


Subject(s)
Humans , Male , Female , Infant, Newborn , Birth Weight , Infant, Low Birth Weight , Prospective Studies , Weight by Height/physiology
20.
Rev. Inst. Med. Trop. Säo Paulo ; 34(1): 55-60, Jan.-Feb. 1992.
Article in English | LILACS | ID: lil-320629

ABSTRACT

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 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.


Subject(s)
Humans , Male , Female , Infant , Child, Preschool , Child , Adolescent , Toxocariasis/diagnosis , Antibody Specificity , Antigens, Helminth/blood , Enzyme-Linked Immunosorbent Assay , Sensitivity and Specificity , Toxocariasis/immunology
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