RESUMO
Human neurocysticercosis (NCC) is a worldwide neglected disease caused by Taenia solium metacestode and responsible for various complications and neurological disorders. This study aimed to evaluate the use of specific immunoglobulin Y (IgY) produced by laying hens immunized with a hydrophobic fraction of Taenia crassiceps metacestodes (hFTc) in NCC diagnosis. Egg yolk IgY antibodies were fractionated, purified and characterized. Enzyme-linked immunosorbent assay (ELISA) was carried out to evaluate the production kinetics and avidity maturation of anti-hFTc IgY antibodies throughout the IgY obtention process. Antigen recognition tests were carried out by Western blotting and immunofluorescence antibody test using purified and specific anti-hFTc IgY antibodies for detection of parasitic antigens of T. crassiceps and T. solium metacestodes. Sandwich ELISA was performed to detect circulating immune complexes formed by IgG and parasitic antigens in human sera. The results showed high diagnostic values (93.2% sensitivity and 94.3% specificity) for immune complexes detection in human sera with confirmed NCC. In conclusion, specific IgY antibodies produced from immunized hens with hFTc antigens were efficient to detect T. solium immune complexes in human sera, being an innovative and potential tool for NCC immunodiagnosis.
Assuntos
Antígenos de Helmintos/imunologia , Imunoglobulinas/sangue , Testes Imunológicos/métodos , Neurocisticercose/parasitologia , Taenia/isolamento & purificação , Animais , Afinidade de Anticorpos , Galinhas , Feminino , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Neurocisticercose/imunologia , Óvulo , Taenia/imunologiaRESUMO
INTRODUCTION: Detection of Strongyloides stercoralis larvae is particularly challenging because only a small number of larvae are released into the feces, regardless of infection stage. OBJECTIVE: Our objective was to apply conventional polymerase chain reaction (PCR) to the detection of S. stercoralis DNA in feces samples to evaluate its performance in samples of patients with strongyloidiasis and compare results with those of immunodiagnosis. METHODS: Stool, serum, and saliva samples were collected from each individual (n = 48) at the clinic hospital of the State University of Londrina, Brazil, for parasitological, immunological, and molecular tests. Stool samples were processed via parasitological methods. Serum samples were used for immunoglobulin G (IgG) detection and saliva samples for IgA detection by ELISA. RESULTS: For amplification by conventional PCR, two different primers were used: species specific (101 bp) and genus specific (392 bp). The results showed that 34 (97.1%) of the 35 copro-positive individuals for S. stercoralis were positive for serum IgG and 19 (54.3%) were positive for salivary IgA. Regarding molecular analysis, both primers (species and genus specific) demonstrated positivity in 100% of the samples, which was confirmed by sequencing the positive samples. CONCLUSION: Complementary examinations of the parasitological method demonstrated excellent results in the context of the diagnosis of strongyloidiasis, especially in asymptomatic patients with irregular larval release in the feces.
Assuntos
Imunoensaio , Técnicas de Diagnóstico Molecular , Strongyloides/genética , Strongyloides/imunologia , Estrongiloidíase/diagnóstico , Estrongiloidíase/parasitologia , Animais , Ensaio de Imunoadsorção Enzimática , Humanos , Imunoglobulina A Secretora , Imunoglobulina G/imunologia , Reação em Cadeia da Polimerase , RNA Ribossômico 18S/genética , Ratos , Saliva/imunologiaRESUMO
Human strongyloidiasis is an infection caused by the helminth Strongyloides stercoralis that can be fatal, especially in immunosuppressed patients. The aim of this study is to evaluate parasite-specific IgG and IgA levels using S. venezuelensis third-stage (L3) infective larvae alkaline extract as a heterologous antigen by ELISA in paired serum and saliva samples with improved sensitivity and specificity. Individuals from northern Paraná state, Brazil were divided into three groups: 30 patients copropositive for S. stercoralis (Group I); 30 clinically healthy individuals (Group II); and 30 patients copropositive for other parasites (Group III). The area under ROC curve (AUC), an overall index of diagnostic accuracy, and Kappa index were calculated. Data were analyzed using analysis of variance (ANOVA) followed by a Kruskal-Wallis test. Probability (p) values of <0.05 were regarded as significant. In Group I, IgG was detected in 96.7% serum and in 6.7% saliva samples. IgG was not detected in Group II. In Group III, cross-reactivity was observed for serum IgG in 26.7% and in 6.7% for saliva samples. In Group I, IgA was detected in 76.7% serum and 56.7% saliva samples. In Group II, 3.3% were positive for IgA in serum, whereas IgA was not detected in any saliva samples. Group III showed 6.7% serum and 26.7% saliva-positive samples. The sensitivity values for detection of IgG and IgA in serum samples were 96.7% and 76.7%, respectively. In saliva samples, the sensitivity values for detection of IgG and IgA were 6.7% and 56.7%, respectively. The specificity value was 100% for the detection of IgG in serum and for detection of IgG and IgA in saliva, and 96.7% for detection of IgA in serum samples. The proper choice of immunological diagnosis to supplement parasitological methods is essential to estimate the true prevalence of the parasite, and will permit analysis of population immune response profiles, particularly in northern Paraná state, where there are no previous reports.
Assuntos
Anticorpos Anti-Helmínticos/análise , Strongyloides stercoralis/imunologia , Estrongiloidíase/diagnóstico , Adolescente , Adulto , Idoso , Animais , Anticorpos Anti-Helmínticos/sangue , Área Sob a Curva , Brasil/epidemiologia , Criança , Pré-Escolar , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Hospedeiro Imunocomprometido , Imunoglobulina A/análise , Imunoglobulina A/sangue , Imunoglobulina G/análise , Imunoglobulina G/sangue , Larva/imunologia , Masculino , Pessoa de Meia-Idade , Saliva/imunologia , Sensibilidade e Especificidade , Soro/imunologia , Estrongiloidíase/epidemiologia , Estrongiloidíase/parasitologia , Adulto JovemRESUMO
This study investigated the water quality in an urban eutrophic reservoir in Northeastern Brazil, considering the influence of seasonality. Monthly, samples were collected in the sub-surface reservoir. The following abiotic variables were analyzed: temperature, pH, dissolved oxygen, apparent color, turbidity, conductivity, fluoride, total nitrogen, chlorides, total dissolved solids, total hardness, iron, copper, manganese, aluminum, chlorophyll-a and phaeophytin. Total and thermotolerant coliforms were analyzed according to APHA (2012). Cyanobacteria density was quantified through its biomass. The data were analyzed using one- way ANOVA and Pearson correlation test. Higher values mean phytoplankton biomass (26.3mm3.L-1) occurred in the dry season, especially Planktothrix agardhii and Geiterinema amphibium, which occurred in 100% of samples. High trophic state index was detected throughout the year. Seasonality exerted some influence on both biotic and abiotc variables, leading to changes in water quality of the reservoir.
Assuntos
Cianobactérias , Monitoramento Ambiental , Água Doce/química , Água Doce/microbiologia , Fitoplâncton , Qualidade da Água , Biomassa , Brasil , Estações do AnoRESUMO
Objetivo. El objetivo de este trabajo fue caracterizar en el biomodelo Mesocricetus auratus la sintomatología y lesiones anatomopatológicas que provocan 5 aislados clínicos de Leptospira spp., provenientes de Nicaragua. Materiales y métodos. Con este fin se inocularon 50 hámster por vía i.p con 1mL del cultivo de cada una de las cepas en fase exponencial teniendo una concentración celular de 7.5 x 106 leptospira/mL (10 animales por cepa), evaluándose signos de la enfermedad, mortalidad durante 14 días, lesiones anatomopatológicas macroscópicas y microscópicas mediante tinción con hematoxilina-eosina y tinción de Warthyn Starryn. Resultados. Todas las cepas presentaron alta mortalidad, mostrando un cuadro tanto clínico, como lesional característico de la infección experimental. Además, causaron la muerte al 100% de los animales entre el tercer y décimo día postinfección. En el estudio anatomopatológico la cepa del serogrupo Ballum y la del serogrupo Pomona produjeron focos de hemorragias específicamente en el riñón y pulmones. De forma similar ocurrió una congestión hepática y renal, mientras que la hemorragia renal fue observada con mayor frecuencia en la cepa del serogrupo Pomona, diferenciándose del resto de las cepas que mostraron esta lesión con menos frecuencia. Conclusiones. Este trabajo permitió una mayor caracterización de estas cepas siendo utilizadas como futuras candidatas vacunales frente a una nueva epidemia de Leptospirosis en Nicaragua.
Objective. The aim of this study was to characterize the symptomatology and anatomopathological lesions caused by 5 clinical isolated Leptospira spp. from Nicaragua in a Mesocricetus auratus biomodel. Materials and methods. 50 hamsters were inoculated via i.p with 1mL of the culture of each strain in exponential phase having a cellular concentration of 7.5 x 106 leptospira/mL, (10 animals per strain). Signs of the disease, mortality during 14 days, and macroscopic and microscopic anatomopathological lesions by haematoxylin-eosin and Warthyn Starryn stain technique were evaluated. Results. All the strains presented high mortality, showing clinical lesions of the experimental infection. Death to 100% of the animals was caused between the third and tenth day post-infection. In the anatomopathologic study, the strains of the Ballum and Pomona serogroup produced haemorrhaging specifically in the kidney and lungs. The animals manifested hepatic and renal congestion, while the renal haemorrhage was observed with more frequency in the strain of the Pomona serogroup, differing from the other strains, which presented this lesion less frequently. Conclusions. This work allowed a better characterization of these strains in order to use them as future vaccine candidates for future Leptospirosis epidemics in Nicaragua.
Assuntos
Humanos , Animais , Leptospira , Mesocricetus , Sinais e SintomasRESUMO
Objetivo. El objetivo de este trabajo fue comparar los índices espontáneos e inducidos en ratones de ambos sexos de las líneas Balb/c, NMRI, OF-1 y C57BL/6/cenp, mediante el ensayo de aberraciones cromosómicas en células de la médula ósea. Materiales y métodos. Con este fin se determinó la línea más eficiente, sobre la base de la aparición significativa de índices espontáneos más bajos e índices inducidos altos a sustancias mutagénicas como la ciclofosfamida (CF). Resultados. Se obtuvo como resultado que la línea Balb/c en ambos sexos difiere significativamente de las demás líneas evaluadas. En esta línea se encontró el menor número de células totales con aberraciones estructurales y un mayor número de inducción de aberraciones cromosómicas cuando es utilizada la CF como mutágeno. Conclusiones. Este trabajo permitirá utilizar la mejor línea de ratones como biomodelo en este ensayo de genotoxicidad, además que le confiere a esta técnica citogenética mayor veracidad y robustez.
Objective. The aim of this study was to compare spontaneous and induced rates in mice of both sexes of lines BALB / c, NMRI, OF-1 and C57BL/6/ cenp, by testing chromosomal aberrations in bone marrow cells. Materials and methods. For this purpose, the most efficient line was determined, based on the significant appearance of lower spontaneous rates and higher induced rates to mutagen substances such as cyclophosphamide (CP). Results. The result obtained was that line Balb / c, in both sexes, differs significantly from the other lines tested. The lesser total number of cells with structural aberrations was found in this line and a greater number of induction of chromosomal aberrations when CP was used as a mutagen. Conclusions. This work will give way to the use of the best line of mice as bio model in this genotoxicity test, and will also give this cytogenetic technique greater veracity and robustness.
Assuntos
Medula Óssea , Aberrações Cromossômicas , CamundongosRESUMO
The aim of the present research was to test the application of Taenia saginata metacestodes as an alternative antigen for use in enzyme-linked immunosorbent assay (ELISA) and Western Blotting (WB) tests compared with the use of metacestodes antigen of Taenia solium in cerebrospinal fluid (CSF) samples. The samples were obtained from 35 patients with definitive neurocysticercosis (NCC)-group 1-and 44 patients with other neurological disorders (control)-group 2. The sensitivity and specificity of ELISA, using antigen obtained from T. solium, applied to the patients of group 1 yielded results of 100%. When the tests were conducted using T. saginata metacestodes, results were 100% and 93.2%, respectively. The 47-52-, 64-68-, and 70-kDa antigens showed high frequencies in CSF samples from patients with NCC when WB was conducted with both antigens. The results indicate that T. saginata metacestodes can be used as an alternative antigen for the diagnosis of human NCC in CSF samples.
Assuntos
Anticorpos Anti-Helmínticos/líquido cefalorraquidiano , Antígenos de Helmintos/isolamento & purificação , Neurocisticercose/diagnóstico , Taenia saginata/química , Taenia solium/química , Adolescente , Adulto , Animais , Western Blotting/métodos , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Humanos , Testes Imunológicos , Masculino , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Taenia saginata/imunologia , Taenia solium/imunologia , Adulto JovemRESUMO
Strongyloidiasis, caused by the nematode Strongyloides stercoralis, is one of the major worldwide parasitic infections in humans. Breastfeeding may offer a potential protection against this infection. Feces, serum and milk samples were obtained from 90 lactating women from Clinical Hospital of Universidade Federal de Uberlândia, Brazil. The fecal samples were collected for parasitological diagnosis and the serum and milk samples were examined for specific S. stercoralis IgA and IgG antibodies using the indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA). Fecal examination showed that the rate of prevalence of S. stercoralis infection in the lactating women was 4.4%. IFAT manifested a 16.7% positivity rate for specific IgA antibody in serum and a 28.9% rate in milk samples; specific IgG was 41.1% in serum and 25.5% in milk samples. According to ELISA the positivity rate for specific IgA antibody was 21.1% in serum and 42.2% in milk samples; specific IgG was 40% in serum and 18.9% in milk samples. In serum samples, these immunological tests showed a concurrence of 91.1% and 94.4%, respectively, in detecting specific IgA and IgG antibodies. In milk samples, they showed a concurrence of 70% and 78.9%, respectively, in detecting specific IgA and IgG antibodies. There was a statistically significant difference between concordant and discordant results of immunological tests (P<0.0001). IFAT and ELISA highly concurred in their detection of specific S. stercoralis IgA and IgG antibodies in serum and in milk samples reconfirming prior studies that the serological method is a complement to the direct diagnosis of the parasite, and suggesting that immunological methods using milk samples can also be helpful. Furthermore, in endemic areas, infants may acquire antibodies to S. stercoralis from breast milk, possibly, contributing to the enhancement of specific mucosal immunity against this parasite.
Assuntos
Anticorpos Anti-Helmínticos/análise , Anticorpos Anti-Helmínticos/sangue , Leite Humano/imunologia , Soro/imunologia , Estrongiloidíase/imunologia , Adolescente , Adulto , Animais , Brasil , Ensaio de Imunoadsorção Enzimática , Fezes/parasitologia , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Imunoglobulina A/análise , Imunoglobulina A/sangue , Imunoglobulina G/análise , Imunoglobulina G/sangue , Strongyloides stercoralis/imunologiaRESUMO
This study examined the frequency of Strongyloides stercoralis infection in patients with gastrointestinal cancer through parasitological and immunological tests. A total of 77 patients were evaluated, 33 with gastrointestinal cancer and 44 controls with other types of cancers. All the patients were undergoing chemotherapy and 14 (18.2%) were receiving concomitant radiotherapy. For a parasitological diagnosis, we applied the Baermann and Lutz methods. The immunological diagnosis involved the indirect fluorescence antibody test (IFAT) and enzyme linked immunosorbent assay (ELISA) to detect IgG antibodies using Strongyloides ratti antigens. The frequency of positive S. stercoralis in gastrointestinal cancer diagnosed by parasitological methods was 3 cases (9.1%), by serology it was 8 cases (24.2%). In the control group 1 case (2.3%) of S. stercoralis was diagnosed by parasitological methods and 2 cases (4.5%) by immunological tests (p<0.05). Patients with gastrointestinal cancer had a 6.7-fold greater chance of testing positive for S. stercoralis infection. Our data highlight the importance of parasitological and immunological diagnosis for S. stercoralis in patients with gastrointestinal cancer living in endemic areas of strongyloidiasis, since they have a higher risk of becoming infected with S. stercoralis than patients with other types of cancer.
Assuntos
Neoplasias Gastrointestinais/imunologia , Hospedeiro Imunocomprometido , Strongyloides stercoralis , Estrongiloidíase/diagnóstico , Adolescente , Adulto , Idoso , Animais , Antineoplásicos/efeitos adversos , Antineoplásicos/imunologia , Brasil , Estudos de Casos e Controles , Fezes/parasitologia , Feminino , Neoplasias Gastrointestinais/tratamento farmacológico , Humanos , Testes Imunológicos , Masculino , Pessoa de Meia-Idade , Parasitologia/métodos , Prevalência , Strongyloides stercoralis/imunologia , Estrongiloidíase/complicações , Estrongiloidíase/imunologiaRESUMO
Leucemias agudas necessitam de suporte hemoterápico para serem tratadas adequadamente. A presença do anticorpo anti-Lutheran B e a alta freqüência do antígeno em nossa população dificulta o tratamento de doenças hematológicas que necessitam de suporte transfusional, tais como a leucemia linfóide aguda (LLA). Alternativas de tratamento hemoterápico são abordadas neste artigo. Descreve-se o caso de uma paciente com LLA e anticorpo anti-Lutheran B. A dificuldade em encontrar sangue compatível levou os autores a adotarem estratégia conservadora no tratamento hemoterápico. O uso de eritropoetina e a adoção de protocolo quimioterápico individualizado viabilizaram os resultados obtidos.
Acute leukemia needs hemotherapeutic support to be successfully treated. The Lutheran B antibody, specifically the high frequency of this antigen in our population, has made it difficult to treat hematological malignancies that need transfusional support, including acute lymphoblastic leukemia. Finding compatible blood is complex and so the authors adopted a conservative treatment strategy. The prescription of epoetin and an individualized chemotherapy protocol may improve results. We describe a case report of a patient with anti-Lutheran B antibody and how difficulties to find compatible blood were solved.
Assuntos
Leucemia-Linfoma Linfoblástico de Células Precursoras B , Incompatibilidade de Grupos Sanguíneos , Transfusão de Sangue , Eritropoetina , Neoplasias Hematológicas , Guias como Assunto , Tratamento Farmacológico , Sistema do Grupo Sanguíneo Lutheran , AntígenosRESUMO
O objetivo deste trabalho é relatar um caso de macroglobulinemia de Waldenstrõm com envolvimento retiniano e da circulação coroidiana, com boa resposta ao tratamento. O desenho é um caso controle intervencional. Um homem de 47 anos foi encaminhado para o serviço de hematologia para avaliação de anemia e linfadenopatia inguinal. O diagnóstico de macroglobulinemia de Waldenstrõm foi feito com base nos achados clínicos e laboratoriais. O paciente desenvolveu hemorragia intra-ocular sem associação com hipertensão e com fatores de coagulação normais. Exame fundoscópico revelou retardamento na circulação coriorretiniana, microaneurismas, e telangiectasias no pólo posterior. Angiografia fluorescente demonstrou o prolongamento do tempo da circulação coroidiana, tempo de trânsito arteriovenoso também prolongado, e áreas de não perfusão capilar. Estes resultados foram acompanhados por uma elevação acentuada da IgM. A maioria dos achados oculares foi revertida com a plasmaférese. A macroglobulinemia de Waldenstrõm pode envolver a retina e a circulação coroidiana, porém pode ser reversível com o tratamento apropriado.
The aim of this work is to report a case of Waldenstrõm's Macroglobulinaemia involving both retinal and choroidal circulation that resolved after treatment. The design is an interventional case report. A 47-year old male was admitted for evaluation of anaemia, and groin lymphadenopathy. A diagnosis of Waldenstrõm's macroglobulinaemia was made based on clinical and laboratory findings. The patient developed intraocular hemorrhage without associated hypertension and with normal coagulation profile. A funduscopic examination revealed marked dilation and beading of the venous system, microaneurysms, and telangiectatic capillary beds in the posterior pole. A fluorescein angiography showed delayed choroidal filling, prolonged arteriovenous transit time and areas of capillary non-perfusion. These findings were accompanied by a severe IgM increase in serum viscosity. The most important ocular findings were reversible after plasmapheresis. Waldenstrõm's macroglobulinaemia can involve both retinal and choroidal circulation. The prominent microvasculopathy is reversible after appropriate treatment.
Assuntos
Humanos , Masculino , Pessoa de Meia-Idade , Viscosidade Sanguínea , Imunoglobulinas , Mieloma Múltiplo , Macroglobulinemia de WaldenstromRESUMO
The purpose of this study was to determine the frequency of association between positive Strongyloides stercoralis serology and diabetes mellitus. A total of 78 diabetic patients and 42 controls were evaluated. For a parasitological diagnosis, Baermann and Hoffman et al.'s methods were applied. The immunological diagnosis involved the indirect fluorescence antibody test, ELISA and Western blotting to detect IgG antibodies. The frequency of positive S. stercoralis serology in diabetics was 23% versus 7.1% in the control group (P<0.05). The odds ratio for diabetics was 3.9 (CI, 1.6-15.9, P<0.05). Diabetic patients with HbA(1c)< or =7 had a greater chance of testing negatively for S. stercoralis infection (OR: 1.5, P>0.05). Provided there are related cases of disseminated strongyloidiasis in diabetics and there is a higher frequency of asymptomaticity of the infection in this group, the immunological screening of these patients at risk could prevent severe and fatal outcomes of the disease.
Assuntos
Diabetes Mellitus/parasitologia , Strongyloides stercoralis/crescimento & desenvolvimento , Estrongiloidíase/complicações , Adulto , Idoso , Animais , Anticorpos Anti-Helmínticos/sangue , Western Blotting , Diabetes Mellitus/sangue , Ensaio de Imunoadsorção Enzimática , Eosinófilos/citologia , Fezes/parasitologia , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Hemoglobinas Glicadas/metabolismo , Humanos , Contagem de Leucócitos , Masculino , Pessoa de Meia-Idade , Estrongiloidíase/sangue , Estrongiloidíase/parasitologiaRESUMO
Comparou-se o emprego de cortes em congelacao de larvas de S. stercoralis e de S. ratti como fonte de antigenos na reacao de imunofluorescencia indireta (RIF) para o diagnostico da estrongiloidiase humana. Os antigenos foram obtidos de coproculturas de fezes humanas e de ratos, respectivamente. Soros de 123 individuos foram analisados sendo 54 de pacientes com estrongiloidiase e 69 controles. Empregou-se conjugado anti IgG humano marcado com fluorescencia nos titulos ideais de 10 para S. stercoralise 100 para S. ratti. A sensibilidade da RIF foi de 94,4 por cento e 92,5 por cento e a especificidade de 94,2 por cento e 97,1 por cento respectivamente para os antigenos de S. stercoralis e S. ratti...