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Master's Dissertation
DOI
10.11606/D.9.2000.tde-27012015-130517
Document
Author
Full name
Alessandra Xavier Pardini
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2000
Supervisor
Committee
Vaz, Adelaide Jose (President)
Livramento, Jose Antonio
Takei, Kioko
Title in Portuguese
Utilização de preparações antigênicas de cisticercos de Taenia crassiceps para pesquisa de anticorpos na neurocisticercose (Taenia solium)
Keywords in Portuguese
Imunologia
Parasitologia
Abstract in Portuguese
O complexo teníase-cisticercose humana representa importante problema sócio-econômico e de Saúde Pública em países em desenvolvimento, incluindo nosso país. A forma mais grave da doença decorre da localização de cisticercos no Sistema Nervoso Central, a neurocisticercose. Devido à dificuldade de obtenção de parasitas a partir de suínos naturalmente infectados estudamos como fonte alternativa, preparações antigênicas de cisticercos de Taenia crassiceps (antígeno heterólogo), para pesquisa de anticorpos anti-cisticercos em líquido cefalorraquiano (LCR) de pacientes com neurocisticercose. Foram estudados os extratos antigênicos de líquido vesicular de cisticercos de Taenia crassiceps (LV-Tcra) e as frações purificadas por Concanavalina A (ConA-Tcra) obtida por coluna de afinidade com lectina e glicoprotéico fracionado (GP-Tcra) obtida a partir do antígeno LV-Tcra em eletroforese preparativa. Os antígenos LV-Tcra, ConA-Tcra e GP-Tcra para a detecção de anticorpos IgG anti-T. solium, foram ensaiados por ELISA em amostras de LCR e por imunoblot em amostras de LCR e soro. Foi utilizado também, kit comercial ELISA com antígeno de T. solium. A sensibilidade e a especificidade obtidas para os antígenos LV-Tcra, ConA-Tcra e GP-Tcra no teste ELISA foram de 100%, com boa reprodutibilidade. Os peptídeos em ordem de freqüência de reatividade para o antígeno LV-Tcra foram: 14-11kD (100%), 62kD (100%), 68kD (100%), 91kD (76%), 25kD (70%), 46KD (64%), 18kD (58%), 43kD (23%), 9-8kD (17%), 56kD (11%) e 32kD (11%). Para o antígeno ConA-Tcra foram identificados os peptídeos, por ordem de freqüência, 14kD (100%), 28kD (66%), 18kD (55%), 46kD (44%), 43kD (22%), 94kD (22%), 103kD (22%). Para o antígeno GP-Tcra, foram identificados somente os peptídeos de 14 - 18kD. Amostras de LCR de pacientes com esquistossomose não apresentaram reatividade com os extratos antigênicos e uma amostra de LCR de paciente com neurossífilis, apresentou forte reatividade eom os peptfdeos de baixo peso molecular≤20kD, para os três antígenos, inclusive no teste ELISA comercial com antígeno de T. solium. Os resultados confirmam que os antígenos de T. crassiceps são importantes fontes alternativas de extratos antigênieos. As frações glicoprotéicas mostraram-se eficientes na detecção de anticorpos anti-T. solium em amostras de LCR e soro de pacientes com neurocisticercose.
Title in English
The use of antigenic preparation of Taenia crassiceps cysticercus detect antibodies in neurocysticercosis (Taenia solium)
Keywords in English
Immunology
Parasitology
Abstract in English
The human taeniasis-cysticercosis complex represents an important socioeconomic and Public Health problem in developing countries, including Brazil. The most severe form of the disease is due to the localization of cysticerci in the Central Nervous System, i.e. neurocysticercosis. Due to the difficulty in obtaining parasites from naturally infected swine, we studied an alternative source consisting of Taenia crassiceps cysticerci (heterologous antigen) for the search of anti-cysticercus antibodies in cerebrospinal fluid (CSF) from patients with neurocysticercosis. We studied the antigenic extracts of vesicular fluid of Taenia crassiceps cysticerci (VF-Tcra) and the purified fractions Concanavalin A (ConA-Tcra) obtained from an affinity column with lectin and fractionated glycoprotein (GP-Tcra) obtained from the VF-Tcra antigen by preparative electrophoresis. The VF-Tcra, ConA-Tcra and GP-Tcra antigens for the detection of IgG antibodies were assayed by ELISA in CSF samples and by immunoblot in CSF and serum samples. A commercial ELISA kit with T. solium antigen was also used. The sensitivity and specificity obtained for the VF-Tcra, ConA-Tcra and GP-Tcra were 100% in the ELISA test, with good reproducibility. The peptides in order of frequency of reactivity with the VF-Tcra antigen were: 14-11kD (100%), 62kD (100%), 68kD (100%), 91kD (76%), 25kD (70%), 46KD (64%), 18kD (58%), 43kD (23%), 9-8kD (17%), 56kD (11%), and 32kD (11%). The following peptides, in order of frequency were identified for the ConA-Tcra antigen: 14kD (100%), 28kD (66%), 18kD (55%), 46kD (44%), 43kD (22%), 94kD (22%), and 103kD (22%). Only peptides of 14-18kD were identified for the GP-Tcra antigen. CSF samples from patients with schistosomiasis did not show reactivity with the antigenic extracts and a CSF sample from a patient with neurosyphilis presented strong reactivity with low molecular weight (≤20kD) peptides for the three antigens also in the commercial ELISA with the T. solium antigen. The results confirm that T. crassicepsantigens are important alternative sources of antigenic extracts. The glycoprotein fractions proved to be efficient in detecting anti-T solium antibodies in CSF and serum samples trom patients with neurocysticercosis.
 
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Publishing Date
2015-01-27
 
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