RESUMEN
The aim of the study was to identify the prevalence of hepatitis B and C seropositivity in pregnant women attended in a public maternity hospital located in Catalao-GO from 2005 to 2009. Descriptive, exploratory study conducted through patients` hospital records. For data analysis, we used SPSS version 18.0. The confidence interval (CI) was calculated using the Person χ² test, considering a significance level of 5% (p <0.05). The prevalence of HBV was 5.64% and HCV 0.098%, predominantly in young pregnant women aged between 20 and 30 years old, single and in their first pregnancy.
Asunto(s)
Hepatitis B/epidemiología , Hepatitis C/epidemiología , Complicaciones Infecciosas del Embarazo/epidemiología , Adolescente , Adulto , Femenino , Hepatitis B/sangre , Hepatitis C/sangre , Humanos , Embarazo , Complicaciones Infecciosas del Embarazo/sangre , Prevalencia , Estudios Seroepidemiológicos , Adulto JovenRESUMEN
The aim of the present study was to analyse Taenia solium metacestode antigens that were derived from the unbound fraction of jacalin affinity chromatography and subsequent tert-octylphenoxy poly (oxyethylene) ethanol Triton X-114 (TX-114) partitioning in the diagnosis of human neurocysticercosis (NCC). Immunoassays were designed to detect T. solium-specific IgG antibodies by ELISA and immunoblot. Serum samples were collected from 132 individuals who were categorised as follows: 40 had NCC, 62 presented Taenia spp or other parasitic diseases and 30 were healthy individuals. The jacalin-unbound (J unbound ) fraction presented higher sensitivity and specificity rates than the jacalin-bound fraction and only this fraction was subjected to subsequent TX-114 partitioning, resulting in detergent (DJ unbound ) and aqueous (AJ unbound ) fractions. The ELISA sensitivity and specificity were 85% and 84.8% for J unbound , 92.5% and 93.5% for DJ unbound and 82.5% and 82.6% for AJ unbound . By immunoblot, the DJ unbound fraction showed 100% sensitivity and specificity and only serum samples from patients with NCC recognised the 50-70 kDa T. solium-specific components. We conclude that the DJ unbound fraction can serve as a useful tool for the differential immunodiagnosis of NCC by immunoblot.
Asunto(s)
Anticuerpos Antihelmínticos/sangre , Antígenos Helmínticos , Taenia solium/inmunología , Animales , Antígenos Helmínticos/inmunología , Estudios de Casos y Controles , Cromatografía de Afinidad , Ensayo de Inmunoadsorción Enzimática , Humanos , Immunoblotting , Pruebas Inmunológicas/métodos , Neurocisticercosis/diagnóstico , Neurocisticercosis/inmunología , Sensibilidad y EspecificidadRESUMEN
The aim of the present study is to investigate genetic polymorphisms in Taenia solium metacestodes from different Brazilian geographical areas and to relate them to antibody recognition in serum samples of neurocysticercosis (NC) patients. Metacestodes were obtained from the Distrito Federal (DF), Bahia, Minas Gerais (MG) and São Paulo (SP) regions of Brazil. Samples of human sera from 49 individuals with NC, 68 individuals with other helminthiasis and 40 healthy volunteers were analysed (157 individuals in total). Antigens were prepared and used in enzyme-linked immunosorbent assay and western blotting assays to detect specific immunoglobulin G antibodies. Genetic distances between metacestode populations were analysed using random amplified polymorphic DNA (RAPD) analysis. Our results show that there was a higher frequency of reactivity in the DF region in the sera from NC patients (p < 0.05), while discrimination between active and inactive NC was seen only in extracts from the MG and SP regions (p < 0.05). Using RAPD, the sample from the DF region presented a greater increase compared to the other regions. A relationship between genetic polymorphisms among T. solium metacestodes from different areas in Brazil and the differences in antibody detection in patients with NC were established.
Asunto(s)
Anticuerpos Antihelmínticos/sangre , Antígenos Helmínticos/inmunología , ADN de Helmintos/genética , Inmunoglobulina G/sangre , Polimorfismo Genético/genética , Taenia solium/genética , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Animales , Western Blotting , Brasil , Estudios de Casos y Controles , Niño , Preescolar , Ensayo de Inmunoadsorción Enzimática , Femenino , Geografía , Humanos , Masculino , Persona de Mediana Edad , Técnica del ADN Polimorfo Amplificado Aleatorio , Taenia solium/inmunología , Taenia solium/aislamiento & purificación , Adulto JovenRESUMEN
OBJECTIVE: To evaluate the detergent and aqueous phases of crude saline extract (crude) of Taenia solium metacestodes extracted with Triton X-114, for the detection of immunoglobulin G (IgG) antibodies by enzyme-linked immunosorbent assay (ELISA) and immunoblot for the laboratory diagnosis of human neurocysticercosis. METHOD: Serum samples were collected from 138 individuals and tested; 40 were diagnosed with neurocysticercosis (NC; group 1), 58 presented other parasitic diseases (group 2) and 40 were apparently healthy (group 3). RESULTS: The detergent phase showed only one dominant component (70-50 kDa), while the aqueous phase showed four (116, 110, 97 and 77 kDa) by silver staining. ELISA sensitivity and specificity were 92.5% and 84.5% for crude phase, 92.5% and 93.3% for detergent phase and 72.5% and 72.6% for aqueous phase. CONCLUSIONS: The immunoblot of the antigens confirmed the results obtained by ELISA. Detergent phase purified from T. solium metacestodes was an important source of specific peptides and was efficient in detecting T. solium metacestodes antibodies in serum samples. This phase can be used for detecting IgG antibodies to T. solium metacestodes in seroepidemiological investigations and in the diagnostic screening of NC patients.
Asunto(s)
Anticuerpos Antihelmínticos/sangre , Antígenos Helmínticos , Inmunoglobulina G/sangre , Neurocisticercosis/diagnóstico , Taenia/inmunología , Animales , Ensayo de Inmunoadsorción Enzimática/normas , Humanos , Immunoblotting/normas , Sensibilidad y EspecificidadRESUMEN
INTRODUCTION: Considering that alternative antigens for diagnosing neurocysticercosis continue to be a challenge because of the increasing difficulty in obtaining parasites from naturally infected pigs for preparation of Taenia solium homologous antigen, the aim of the present study was to evaluate the detergent (D) and aqueous (A) fractions from saline extract of Taenia saginata metacestodes for diagnosing neurocysticercosis. METHODS: Taenia saginata was obtained from naturally infected bovines in the Triângulo Mineiro region, State of Minas Gerais, Brazil. The carcasses came from cold storage units and had been slaughtered in accordance with the inspection technique recommended by the Federal Inspection Service. The D and A fractions were obtained by using Triton X-114 (TX-114). Serum samples were obtained from 40 patients with a diagnosis of neurocysticercosis, 45 with other parasitic diseases and 30 from apparently normal individuals. IgG antibody levels were evaluated using the ELISA and immunoblotting assays. RESULTS: The ELISA sensitivity and specificity were 95% and 73.3%, when using saline extract; 95% and 82.6% for the D fraction; and 65% and 61.3% for the A fraction, respectively. The immunoblotting assay confirmed the ELISA results, such that the D fraction was more efficient than the other extracts, and the 70-68 kDa component was immunodominant among neurocysticercosis patients. CONCLUSIONS: These results demonstrated that the D fraction from Taenia saginata metacestodes obtained using TX-114 can be used as a heterologous antigenic fraction in the immunoblotting assay for serologically diagnosing human neurocysticercosis, given its ability to select immunodominant antigens.
Asunto(s)
Antígenos Helmínticos , Neurocisticercosis/diagnóstico , Taenia saginata/inmunología , Taenia solium/inmunología , Animales , Antígenos Helmínticos/aislamiento & purificación , Biomarcadores , Estudios de Casos y Controles , Bovinos , Electroforesis en Gel de Poliacrilamida , Ensayo de Inmunoadsorción Enzimática , Humanos , Immunoblotting , Inmunoglobulina G/sangre , Sensibilidad y Especificidad , Taenia saginata/químicaRESUMEN
The aim of the study was to identify the prevalence of hepatitis B and C seropositivity in pregnant women attended in a public maternity hospital located in Catalao-GO from 2005 to 2009. Descriptive, exploratory study conducted through patients` hospital records. For data analysis, we used SPSS version 18.0. The confidence interval (CI) was calculated using the Person χ² test, considering a significance level of 5% (p <0.05). The prevalence of HBV was 5.64% and HCV 0.098%, predominantly in young pregnant women aged between 20 and 30 years old, single and in their first pregnancy.
El objetivo de este estudio fue identificar la prevalencia de seropositividad para los virus de la hepatitis B y C en mujeres embarazadas atendidas en una maternidad pública de Catalão-GO entre los años 2005 y 2009. Se trata de un estudio descriptivo-exploratorio realizado a partir de la revisión de las fichas clínicas de las pacientes. Para el análisis de los datos se utilizó el programa SPSS versión 18.0. El intervalo de confianza (IC) fue calculado por medio del test de Person χ², considerando un nivel de significancia de 5% (p <0,05). Las tasas de prevalencia de los virus de la hepatitis B y C fueron de 5,64% y 0,098% respectivamente. La mayor frecuencia de seropositividad de estos virus se observó en embarazadas entre 20 y 30 años, solteras y primigestas.
O objetivo do estudo foi identificar a prevalência de soropositividade para as hepatite B e C em gestantes atendidas em uma maternidade pública do município de Catalão-GO no período de 2005 a 2009. Estudo descritivo, exploratório, realizado por meio de consulta aos prontuários. Para a análise dos dados utilizou-se SPSS versão 18.0. O intervalo de confiança (IC) foi calculado por meio do teste de Person χ², considerando um nível de significância de 5% (p< 0,05). A taxa de prevalência de VHB foi de 5,64% e a de VHC, de 0,098%, predominantemente em gestantes jovens, com idade entre 20 e 30 anos, solteiras e primigestas.
Asunto(s)
Adolescente , Adulto , Femenino , Humanos , Embarazo , Adulto Joven , Hepatitis B/epidemiología , Hepatitis C/epidemiología , Complicaciones Infecciosas del Embarazo/epidemiología , Hepatitis B/sangre , Hepatitis C/sangre , Complicaciones Infecciosas del Embarazo/sangre , Prevalencia , Estudios SeroepidemiológicosRESUMEN
The aim of the present study was to analyse Taenia solium metacestode antigens that were derived from the unbound fraction of jacalin affinity chromatography and subsequent tert-octylphenoxy poly (oxyethylene) ethanol Triton X-114 (TX-114) partitioning in the diagnosis of human neurocysticercosis (NCC). Immunoassays were designed to detect T. solium-specific IgG antibodies by ELISA and immunoblot. Serum samples were collected from 132 individuals who were categorised as follows: 40 had NCC, 62 presented Taenia spp or other parasitic diseases and 30 were healthy individuals. The jacalin-unbound (J unbound ) fraction presented higher sensitivity and specificity rates than the jacalin-bound fraction and only this fraction was subjected to subsequent TX-114 partitioning, resulting in detergent (DJ unbound ) and aqueous (AJ unbound ) fractions. The ELISA sensitivity and specificity were 85% and 84.8% for J unbound , 92.5% and 93.5% for DJ unbound and 82.5% and 82.6% for AJ unbound . By immunoblot, the DJ unbound fraction showed 100% sensitivity and specificity and only serum samples from patients with NCC recognised the 50-70 kDa T. solium-specific components. We conclude that the DJ unbound fraction can serve as a useful tool for the differential immunodiagnosis of NCC by immunoblot.
Asunto(s)
Animales , Humanos , Anticuerpos Antihelmínticos/sangre , Antígenos Helmínticos , Taenia solium/inmunología , Antígenos Helmínticos/inmunología , Estudios de Casos y Controles , Cromatografía de Afinidad , Ensayo de Inmunoadsorción Enzimática , Immunoblotting , Pruebas Inmunológicas/métodos , Neurocisticercosis/diagnóstico , Neurocisticercosis/inmunología , Sensibilidad y EspecificidadRESUMEN
The aim of the present study is to investigate genetic polymorphisms in Taenia solium metacestodes from different Brazilian geographical areas and to relate them to antibody recognition in serum samples of neurocysticercosis (NC) patients. Metacestodes were obtained from the Distrito Federal (DF), Bahia, Minas Gerais (MG) and São Paulo (SP) regions of Brazil. Samples of human sera from 49 individuals with NC, 68 individuals with other helminthiasis and 40 healthy volunteers were analysed (157 individuals in total). Antigens were prepared and used in enzyme-linked immunosorbent assay and western blotting assays to detect specific immunoglobulin G antibodies. Genetic distances between metacestode populations were analysed using random amplified polymorphic DNA (RAPD) analysis. Our results show that there was a higher frequency of reactivity in the DF region in the sera from NC patients (p < 0.05), while discrimination between active and inactive NC was seen only in extracts from the MG and SP regions (p < 0.05). Using RAPD, the sample from the DF region presented a greater increase compared to the other regions. A relationship between genetic polymorphisms among T. solium metacestodes from different areas in Brazil and the differences in antibody detection in patients with NC were established.
Asunto(s)
Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Animales , Niño , Preescolar , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Anticuerpos Antihelmínticos/sangre , Antígenos Helmínticos/inmunología , ADN de Helmintos/genética , Inmunoglobulina G/sangre , Polimorfismo Genético/genética , Taenia solium/genética , Western Blotting , Brasil , Estudios de Casos y Controles , Ensayo de Inmunoadsorción Enzimática , Geografía , Técnica del ADN Polimorfo Amplificado Aleatorio , Taenia solium/inmunología , Taenia solium/aislamiento & purificaciónRESUMEN
INTRODUCTION: Considering that alternative antigens for diagnosing neurocysticercosis continue to be a challenge because of the increasing difficulty in obtaining parasites from naturally infected pigs for preparation of Taenia solium homologous antigen, the aim of the present study was to evaluate the detergent (D) and aqueous (A) fractions from saline extract of Taenia saginata metacestodes for diagnosing neurocysticercosis. METHODS: Taenia saginata was obtained from naturally infected bovines in the Triângulo Mineiro region, State of Minas Gerais, Brazil. The carcasses came from cold storage units and had been slaughtered in accordance with the inspection technique recommended by the Federal Inspection Service. The D and A fractions were obtained by using Triton X-114 (TX-114). Serum samples were obtained from 40 patients with a diagnosis of neurocysticercosis, 45 with other parasitic diseases and 30 from apparently normal individuals. IgG antibody levels were evaluated using the ELISA and immunoblotting assays. RESULTS: The ELISA sensitivity and specificity were 95 percent and 73.3 percent, when using saline extract; 95 percent and 82.6 percent for the D fraction; and 65 percent and 61.3 percent for the A fraction, respectively. The immunoblotting assay confirmed the ELISA results, such that the D fraction was more efficient than the other extracts, and the 70-68kDa component was immunodominant among neurocysticercosis patients. CONCLUSIONS: These results demonstrated that the D fraction from Taenia saginata metacestodes obtained using TX-114 can be used as a heterologous antigenic fraction in the immunoblotting assay for serologically diagnosing human neurocysticercosis, given its ability to select immunodominant antigens.
INTRODUÇÃO: Considerando que antígenos alternativos para o diagnóstico da neurocisticercose (NC) continua sendo um desafio devido ao aumento da dificuldade em se obter parasitas de suínos naturalmente infectados, para a preparação do antígeno homólogo de Taenia solium, o objetivo do presente estudo foi avaliar frações detergente (D) e aquosa (A), do extrato salino de metacestódeo de Taenia saginata para diagnóstico da NC. MÉTODOS: Bovinos, naturalmente infectados com Taenia saginata, procedentes da região do Triângulo Mineiro, Estado de Minas Gerais, Brasil, foram obtidos de frigoríficos e abatidos de acordo com a técnica de inspeção recomendada pelo Serviço de Inspeção Federal. As frações D e A foram obtidas utilizando Triton X-114 (TX-114). Amostras de soro foram obtidas de 40 pacientes com diagnóstico de NC, 45 com diagnóstico de outras doenças parasitárias e 30 de indivíduos aparentemente normais. Níveis de IgG foram avaliados pelos testes ELISA e Imunoblotting. RESULTADOS: A sensibilidade e especificidade do teste ELISA foram 95 por cento e 73,3 por cento, quando utilizado o extrato salino, 95 por cento e 82,6 por cento para fração D, e 65 por cento e 61,3 por cento para a fração A, respectivamente. O ensaio Imunoblotting confirmou os resultados do teste ELISA, sendo a fração D mais eficiente que os outros extratos, observando-se que o componente 70-68kDa se comportou como imunodominante para os pacientes com NC. CONCLUSÕES: Estes resultados demonstraram que a fração D de metacestódeo de Taenia saginata obtida com TX-114 pode ser utilizada como fração antigênica heteróloga pelo Imunoblotting para o diagnóstico sorológico da NC humana, considerando sua habilidade para selecionar antígenos imunodominantes.