Rendimiento diagnóstico del Western Blot para detectar simultaneamente anticuerpos en pacientes con cisticercosis, hidatidosis y fascioliasis humana / Western Blot diagnostic yield for simultaneous antibody-detection in patients with human cysticercosis, hydatidosis, and human fascioliasis

RESUMEN Objetivo. Determinar el rendimiento diagnóstico de la técnica de Western Blot para detectar simultáneamente anticuerpos en pacientes con cisticercosis, hidatidosis y fascioliasis humana. Materiales y métodos. Estudio transversal de evaluación de prueba diagnóstica. Se obtuvieron los antígenos de excreción-secreción de las larvas de Taenia solium, quistes de Echinococcus granulosus; y la forma adulta de Fasciola hepática; que luego fueron separados electroforéticamente en geles de poliacrilamida individuales, transferidos y fijados a una membrana de nitrocelulosa para ser enfrentados con sueros de pacientes con las tres parasitosis. La sensibilidad de la técnica se evaluó empleando 300 sueros individuales, 60 pools de dos parasitosis y 20 pools de tres parasitosis y la especificidad con 75 sueros de pacientes con otras parasitosis, 10 de pacientes con otras enfermedades y 15 sueros de personas no parasitadas. Resultados . La técnica reconoció trece glicoproteínas (GP): GP 35, 31, 24, 23, 18, 17, 14 y 13 kDa para cisticercosis, GP 8,16 y 21 kDa para hidatidosis y GP: 17 y 23 kDa para fascioliasis. La prueba detectó la presencia de anticuerpos alcanzando una sensibilidad de 96% (IC95%: 94,62-98,54%) en la detección de una o las trece bandas, una especificidad de 100% (IC95%: 99,50 - 100,00%); individualmente, se tuvo una sensibilidad para cisticercosis de 97% (IC95%: 93,16-100%), para hidatidosis de 94% (IC95%: 88,85-99,15%) y para fascioliasis de 96% (IC95%: 91,66-100%). Conclusiones. La prueba de Western blot es eficaz en la detección, simultanea de anticuerpos en pacientes con cisticercosis, hidatidosis y fascioliasis humana, y puede ser utilizada como prueba de descarte o confirmatoria en zonas endémicas.

ABSTRACT Objectives . To determine the diagnostic yield using western blotting to simultaneously detect antibodies in patients with human cysticercosis, hydatidosis, and human fascioliasis. Materials and methods . Cross-sectional study of diagnostic yield assessment. Excretory/secretory antigens were obtained from Taenia solium larvae, Echinococcus granulosus cysts, and the adult flukes of Fasciola hepática, which were then separated using the polyacrylamide gel electrophoresis technique, transferred, and attached to a nitrocellulose membrane to be probed with sera from the patient infected with the three parasites. The sensitivity of the technique was assessed using 300 individual serum samples, 60 pools of two parasites, and 20 pools of three parasites with 75 sera from patients with other parasites, 10 from patients with other diseases, and 15 from patients without parasites. Results . The technique revealed 13 glycoproteins (GP): GP 35, 31, 24, 23, 18, 17, 14, and 13 kDa for cysticercosis; GP 8, 16, and 21 kDa for hydatidosis; and GP 17 and 23 kDa for fascioliasis. The test detected the presence of antibodies with a sensitivity of 96% (95% confidence interval [CI] = 94.62-98.54%) in the detection of one or the thirteen bands, a specificity of 100% (95% CI = 99.50-100.00%); individually, there was a sensitivity for cysticercosis of 97% (95% CI = 93.16-100.00%), for hydatidosis of 94% (95% CI = 88.85-99.15%) and for fascioliasis of 96% (95% CI = 91.66-100.00%). Conclusions . Western blotting is effective in the simultaneous detection of antibodies in patients with human cysticercosis, hydatidosis, and fascioliasis, and it can be used as a diagnostic test to either rule out or confirm the presence of antibodies in endemic areas.

Effectiveness of the Live Births Information System in the Far-Western Brazilian Amazon / Eficácia do Sistema de Informação de Nascidos Vivos na Amazônia Extremo-Ocidental Brasileira

The Live Birth Information System (SINASC) was implemented in 1990 for the purpose of providing information about the live-birth characteristics for the establishment of specific health indicators. This work evaluates the information quality of SINASC in relation to its data completeness and coverage for five municipalities from the State of Acre from 2005 to 2010. Lack of information (not filled out or stated as "unknown") was estimated for each variable. Coverage was estimated comparing the Civil Register office statistics in accordance with the mother's municipality of residence. An increase in incompleteness of the majority of variables was observed, and also a decrease in coverage between 2005 and 2010 in these municipalities. These findings do not tally with results from the majority of studies that use SINASC as a data source. The results of this work highlight the relevance of continuous capacity building and the incentive for accurate and complete data inclusion, as well as awareness of the importance of SINASC for public health policies.

O Sistema de Informação de Nascidos Vivos (SINASC) foi implantado no ano de 1990 com o objetivo de fornecer dados sobre as características de nascidos vivos para o estabelecimento de indicadores de saúde específicos. Objetivo: O presente trabalho avalia a qualidade da informação do SINASC quanto à incompletude dos seus dados e da cobertura para cinco municípios do estado do Acre nos anos de 2005 e 2010. Métodos: Foi calculada a incompletude (definida como dados em branco/ignorado) de cada variável, assim como a cobertura desse sistema através da comparação com as estatísticas do Registro Civil, segundo município de residência da mãe. Resultados: Observou-se um aumento da incompletude da maioria das variáveis e uma diminuição da cobertura de 2005 para 2010 no conjunto dos municípios avaliados, destoando dos resultados obtidos na maioria dos estudos que utilizam o SINASC como fonte de dados. Conclusões: Os resultados deste trabalho apontam para a importância da contínua capacitação e também para o incentivo ao preenchimento dos dados de forma correta e completa, bem como a conscientização da importância do SINASC para as políticas públicas de saúde.

Changing Patterns of Serum and Bile Antibodies in Re-infected Rats with Clonorchis sinensis

Rats develop strong resistance to re-infection and super-infection by Clonorchis sinensis. The present study investigated the antibodies present in the sera and bile juice of rats that were primary infected and re-infected with C. sinensis. The serum level of specific IgG antibodies, which were elevated 2 wk of the primary infection, peaked at 4 wk and subsequently remained unchanged even during re-infection. The total IgE level in serum increased slowly from 388 ng / ml to 3,426 ng / ml beginning 2 wk after the primary infection, and remained high up to 8 wk but dropped to a normal level (259 ng / ml) after treatment. In resistant re-infected rats, the serum IgE level increased rapidly and peaked within 1 wk, whereas no increase was observed in immunosuppressed rats. The serum level of specific IgA antibodies was elevated beginning 1 wk after infection, and decreased 4 wk after treatment. The total bile IgA level unchanged during the primary infection but increased in treated and re-infected rats. The elevated levels of serum IgE and bile IgA indicate that these immunoglobulins may be correlated with the development of resistance to re-infection by C. sinensis in rats.

Changing Patterns of Serum and Bile Antibodies in Re-infected Rats with Clonorchis sinensis

Rats develop strong resistance to re-infection and super-infection by Clonorchis sinensis. The present study investigated the antibodies present in the sera and bile juice of rats that were primary infected and re-infected with C. sinensis. The serum level of specific IgG antibodies, which were elevated 2 wk of the primary infection, peaked at 4 wk and subsequently remained unchanged even during re-infection. The total IgE level in serum increased slowly from 388 ng / ml to 3,426 ng / ml beginning 2 wk after the primary infection, and remained high up to 8 wk but dropped to a normal level (259 ng / ml) after treatment. In resistant re-infected rats, the serum IgE level increased rapidly and peaked within 1 wk, whereas no increase was observed in immunosuppressed rats. The serum level of specific IgA antibodies was elevated beginning 1 wk after infection, and decreased 4 wk after treatment. The total bile IgA level unchanged during the primary infection but increased in treated and re-infected rats. The elevated levels of serum IgE and bile IgA indicate that these immunoglobulins may be correlated with the development of resistance to re-infection by C. sinensis in rats.

Immunoparasitological diagnosis of Strongyloides stercoralis in garbage collectors in Uberlândia, MQ Brazil

The objective of this study was to determine the presence of Strongyloides stercoralis in urban garbage collectors through the use of immunological and parasitological methods. A total of 92 individuals were evaluated from August, 1997, to June, 1998. For the parasitological diagnosis Baermann and Lutz' methods were applied. The immunological diagnosis involved the indirect fluorescence antibody test (IFAT) and the enzyme-linked immunosorbent assay (ELISA) to detect specific IgG antibodies. Of the 92 workers examined, six (6.5 percent) were infected with larvae of S. stercoralis. The IFAT detected 19 (16.3 percent) and the ELISA 17 (18.5 percent) positive serum samples. The differences between the results of parasitological and immunological methods were statistically significant (p<0.05). These results demonstrate that there is a need to improve the health conditions of this category of city employees.

Ocular toxocariasis in a child: a case report from Kashmir, north India.

Toxocariasis is an important zoonotic disease caused by the second stage larva of Toxocara canis or Toxocara cati . The typical clinical syndromes of toxocariasis in humans are visceral and ocular toxocariasis. Ocular toxocariasis may presents as peripheral inflammatory mass, posterior pole granuloma and endophthalmitis. We report a serologically confirmed case of ocular toxocariasis in 12-year-old female. The diagnosis was confirmed by detection of anti- Toxocara antibodies in aqueous and vitreous sample by enzyme-linked immunosorbent assay. We suggest that ophthalmologist in this region should include ocular toxocariasis in differential diagnosis particularly in children and young adults.

Optical coherence tomography in presumed subretinal Toxocara granuloma: case report

Estudar os achados em tomografia de coerência óptica num caso de granuloma por Toxocara. Paciente com uma lesão macular cicatricial no olho esquerdo foi submetido a retinografia e tomografia de coerência óptica. Nas imagens de tomografia de coerência óptica, o granuloma aparece como uma imagem de uma massa arredondada hiper-reflectiva acima do epitélio pigmentar e abaixo da retina neurossensorial acompanhado de duas lesões satélites menores. A tomografia de coerência óptica pode nos ajudar a melhorar o entendimento da fisiopatologia do granuloma retiniano do Toxocara, seu diagnóstico e conduta.

Abdominal angiostrongyliasis: a case with severe evolution

É relatado um caso de abdome agudo causado por angiostrongilíase abdominal. Um paciente de 42 anos, previamente hígido, apresentou queixa de nove dias de dor abdominal, constipação, disúria, febre e uma massa palpável na fossa ilíaca direita. Uma laparotomia exploradora foi realizada. Após o tratamento cirúrgico o paciente apresentou graves complicações.

Specific IgG antibody subclasses to Angiostrongylus cantonensis in patients with angiostrongyliasis.

Total IgG, IgG1, IgG2, IgG3, IgG4, IgA and IgM specific antibodies against Angiostrongylus cantonensis somatic antigen were determined by enzyme-linked immunosorbent assay (ELISA) in sera from proven human angiostrongyliasis (PA) cases, clinically suspected angiostrongyliasis cases with eosinophilic meningitis (EM) and healthy control (HC). The specific IgA antibody in each of the patient groups was significantly higher than those of the HC group (p < 0.05). The mean ELISA value of the specific IgM in the PA group was not significantly different from that of the HC group (p > 0.05). However, the mean specific IgM ELISA value in the EM group was significantly higher than that of the HC group (p < 0.05). The levels of the specific IgG and IgG subclasses in both patient groups were significantly higher than in the healthy control (HC) group (p < 0.001). Major differences were evident in the distribution of the IgG subclass antibodies between the patient groups. The IgG1 antibody demonstrated the highest sensitivity and specificity while the IgM and IgA responses were generally poor in both patient groups. The levels of the specific IgG antibody subclasses possibly explain immune responses to the parasite.

Immunological Responses of Dogs Experimentally Infected with Dirofilaria immitis

Three dogs were experimentally infected with Dirofilaria immitis. All dogs were euthanised at 30, 36 and 37 weeks after inoculation of D. immitis for the recovery of adult worms. Three cases accounted to 42.91 % recovery of inoculated worms. Serum samples from dogs experimentally inoculated with D. immitis were analyzed by ELISA and immunoblotting methods. Antibody titers of dogs detected by ELISA peaked between 7 and 14 weeks then decreased between weeks 15 to 24 followed by another increase during weeks 25 to 30 and persisted throughout the remainde of the experiment period. Analysis of adult D. immitis protein stained with Coomassie brilliant blue R-250 indicated separately more than 10 bands, and the major bands were 22, 40, 46, 56, 70, 72 and 89 kDa. Antigenic identification of extracts antigens of adults D. immitis by immunoblotting analysis revealed several bands from pooled sera of patent infection (30 weeks after inoculation). The detected bands were 24, 70, 80 and 110 kDa, 22, 72 and 84 kDa, and 58 and 72 kDa in dogs 1, 2 and 3, respectively. Results of antibody titers reached high levels on the 4th molting stage after inoculation of infective larva (L3), and reinforced previous findings that high molecular weight regions are detected in young animals.

Immunoblot evaluation of the specificity of the 29-kDa antigen from young adult female worms Angiostrongylus cantonensis for immunodiagnosis of human angiostrongyliasis.

The antigenic components of Angiostrongylus cantonensis young adult female worm somatic extract (FSE) were revealed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. The sera tested were from patients with proven angiostrongyliasis, other parasitic diseases, and healthy adults. Both the sera and cerebrospinal fluid (CSF) were tested from patients with clinical angiostrongyliasis. The CSF from patients with other neurological diseases were also included. Using SDS-PAGE, we found that the FSE comprised more than 30 polypeptides. Immunoblot analysis revealed at least 12 or 13 antigenic bands in patients with proven or clinical angiostrongyliasis, respectively. The patterns of reactivity recognized by the serum and CSF antibodies against FSE were similar. These antigenic components had molecular masses ranging from less than 14.4 to more than 94 kDa. The prominent antigenic band of 29-kDa might serve as a reliable marker for the diagnosis of angiostrongyliasis. The sensitivity, specificity, positive and negative predictive values of immunoblot analysis in this antigenic band were 55.6%, 99.4%, 83.3% and 97.4%, respectively.

Seroprevalence of Schistosoma mansoni in Puerto Ricans with inflammatory bowel disease

BACKGROUND: The etiology of Inflammatory Bowel Diseases, Crohn's disease (CD) and ulcerative colitis (UC), is unknown. These diseases have a higher incidence in industrialized countries and their pathogenesis involves an over-reaction of the immune system. A genetic factor is believed to predispose to the development of chronic inflammation in response to an unidentified stimulus. Exposure to infections in childhood may modulate future immune responses. Parasitosis, particularly Schistosomiasis, stimulate Th2 immune responses. It has been hypothesized that the absence of these parasitic infections, as seen in economically developed countries, favors a Th1 response that may result in the clinical appearance of Crohn's disease later in life. OBJECTIVE: To determine the prevalence of Schistosoma mansoni antibodies in Puerto Ricans with Inflammatory Bowel Disease and controls. METHODS: Serum from 92 Puerto Ricans with IBD and 106 controls was screened for S. mansoni adult microsomal antigens (MAMA) using the FAST:ELISA assay. Those positive were confirmed with an enzyme-linked immunoelectrotransfer blot test. RESULTS: Seven serum samples (3 UC and 4 controls) were positive for S. mansoni antibodies. There was no significant difference between groups in gender, municipality of origin or seroprevalence of Schistosomiasis. The control group was slightly older than the IBD group. CONCLUSIONS: Our study did not demonstrate an inverse relation between Schistosomiasis and IBD. However, the decreasing prevalence of Schistosomiasis in the general population of Puerto Rico may account for this result.

Toxocariasis con afectación hepática / Toxocariasis with liver involvement

We are reporting 16 cases of toxocariasis found in a two year period. Mean age was 2 years and 9 months. Sex distribution was 1:1. Thirteen (81 per cent) children presented pica, 8 (50 per cent) had pets at home, 10 (62.5 per cent presented anemia and long standing fever, and all eosinophilic leukocytosis. Fundoscopy was normal in all. Toxocara antibodies (Through ELISA) were increased in all of them. High resolution ultrasonography revealed hypoechoic areas in the liver in 50 per cent of the cases. Therapeutic response was good, the clinical signs and symptoms disappearing at the end of treatment. The eosinophilic leukocytosis, ELISA serum positivity for toxocara and ultrasound findings persisted approximately for a year. Toxocariasis is a common parasitosis in our setting. It must be regarded as the first diagnosis when confronted with eosinophilic leukocytosis and abnormal liver findings by ultrasound.

Albendazole therapy for a recurrent orbital hydatid cyst.

Till recently, the treatment of a multiloculated hydatid cyst in the confines of the orbit was every ophthalmologist's nightmare. Over the last decade, two benzimidazole compounds, mebendazole and albendazole, have been tested clinically for use in the chemotherapy of hydatid disease.

Asma y toxocarosis encubierta / Asthma and covert toxocariasis

Se estudió la relación entre asma bronquial y toxocarosis encubierta. Se seleccionaron 38 pacientes con síndrome de asma bronquial. Se estabelecieron criterios de inclusión/exclusión. Como población control se evaluaron 44 pacientes sin asma con los mismos criterios. Los anticuerpos antitoxocara de tipo IgG e IgE se detectaron mediante enzimoinmunoanálisis (ELISA). Las pruebas cutáneas se realizaron mediante inyección de alergenos habituales y de material excretor/secretor de Toxocara canis obtenido por cultivo in vitro de larvas en estadio II (Ag E/S). Dentro de la población con asma resultó serológicamente positiva el 68.42 por ciento, diferenciándose significativamente de la población control (13.63 por ciento). El porcentaje de pacientes asmáticos con ambos marcadores antitoxocara positivos fue 26.31 por ciento. En la población control fue 4.54 por ciento. El 100 por ciento de los pacientes con asma y seropositividad para IgE antitoxocara dieron reactividad cutánea para el Ag E/S de T. canis. Se concluye que el grupo de pacientes con asma bronquial presentó una indudable asociación con los marcadores serológicos antitoxocara IgE e IgG positivos y con la reactvidad cutánea al Ag E/S, por lo que podría inferirse que cursan una toxocarosis encubierta.

Imunodiagnóstico da esquistossomose mansônica com baixa carga parasitária / Immunodiagnosis of schistosomiasis mansonsi with light worm burden

Presently, the schistosomiasis mansoni with low worm burden is frequent, thus immunologic assays of interest for the field diagnosis of Schistosoma mansoni light infections were evaluated here. Assays not assessed before (group I) and those requiring better validation (group II) for the screening of light infections were included in this study. In the group I, the immunofluorescence assays for the detection of IgM antibodies to worm antigens (IgM IFAw) and IgG antibodies to egg antigens (IgG IFAe) gave high levels of sensitivity, specificity, efficiency and predictive value of positive. However, the immunoenzymatic assays for the detection of IgM antibodies to worm antigens (IgM ELISAw) and to egg antigens (IgM ELISAe) had lower levels than the former assays. The assays from the group II designed mostly for the detection of IgG antibodies to same parasite antigens showed good diagnostic performance. The data obtained here contributed to evidenciate at least three category of immunoassays, and we concluded that those from the category I are suitable for seroepidemiologic purposes by keeping their diagnostic features unchanged even varying significantly the intensity of S. mansoni infection.

Antibodies in serum of patients with clonorchiasis before and after treatment.

Sera of 31 patients infected with Clonorchis sinensis were examined using fraction 1 antigen by ELISA during a one-year observation. The results of ELISA with Igs, IgG and IgA demonstrated high sensitivity (100%, 100% and 90%) and specificity (100%, 100% and 87%). Sera specific Igs and IgG were significantly decreased in the 3rd month after treatment with praziquantel (25mg/kg body weight in one dose), and IgA was significantly decreased in the 1st month (paired t-test, p < 0.05). No eggs were found in the stool after treatment. Detection of sera specific Igs, IgG and IgA by ELISA was combined with stool examination to evaluate the effect of praziquantel and the completeness of the cure.

Detection by immunoblot assay of antibodies to Taenia solium cysticerci in sera from residents of rural communities and from epileptic patients in Bali, Indonesia.

Several studies from Bali have indicated the presence of Taenia solium. Relatively little has been reported, however, implicating human exposure to this parasite on Bali based upon the prevalence of anti-T. solium antibodies in asymptomatic and epileptic individuals. This study was conducted to determine by immunoblot assay and ELISA the frequency of anti-cysticercus antibodies in two groups of Balinese. Among 746 residents of four ecologic zones, 94 (13%) were positive by immunoblot. Of 74 epileptic patients from throughout the island, 10 (14%) were positive by immunoblot and 8 (11%) by ELISA; however, only 4 (22%) of the 18 sera positive in either test were positive in both assays. The previously defined high specificity and sensitivity of immunoblot indicates that T. solium cysticercosis is well established in Bali and that a significant amount of epilepsy may be due to neurocysticercosis.