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Master's Dissertation
DOI
10.11606/D.9.2007.tde-12042007-152310
Document
Author
Full name
Roberta Morozetti Blanco
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2007
Supervisor
Committee
Takei, Kioko (President)
Romero, Eliete Caló
Vaz, Adelaide José
Title in Portuguese
Avaliação da glicolipoproteína como antígeno para sorodiagnóstico da leptospirose
Keywords in Portuguese
Dot-ELISA
Glicolipoproteína
GLP
Imunodiagnóstico
Leptospirose
Abstract in Portuguese
Este trabalho visa investigar a resposta sorológica para glicolipoproteína (GLP) de leptospiras com a finalidade de padronizar o uso deste antígeno em testes sorológicos para o diagnóstico da leptospirose humana. Dentre as proteínas envolvidas na patogenicidade das leptospiras, a GLP ainda não foi estudada quanto a imunogenicidade e quanto ao seu emprego como antígeno na detecção de anticorpos específicos. Assim, dot-ELISA para detecção de anticorpos IgG e IgM, com o emprego de GLP de leptospiras patogênica e não patogênica, foi padronizado. Foram testadas amostras pareadas de soros de 90 pacientes com leptospirose confirmada sorologicamente por MAT, sendo as primeiras amostras colhidas na fase aguda da doença e as segundas amostras após 10 a 15 dias. Foram testadas também amostras únicas de 30 pacientes de diferentes patologias, que apresentaram resultados negativos no MAT, pertencentes ao grupo controle. A detecção de anticorpos IgG não mostrou resultados satisfatórios, tendo sido, o seu estudo, descontinuado. Os resultados do dot-ELISA IgM mostraram sensibilidade de 100% nas amostras colhidas após soroconversão no MAT e a precocidade de detecção foi demonstrada pela alta positividade nas amostras colhidas na fase aguda da doença (76,6% para dot-ELISA com antígeno de Leptospira interrogans sorovar Copenhageni e 90% com antígeno de Leptospira biflexa sorovar Patoc). A especificidade foi alta (96,6%), porém o dot-ELISA utilizando ambos os antígenos apresentou 3,3% de resultados falso-positivos. Este estudo demonstrou a importância da resposta imune humoral ao antígeno GLP de leptospiras. A utilização da GLP, no teste dot-ELISA para detecção de anticorpos IgM permitiu realizar o diagnóstico sorológico de forma simples, rápida e com alta eficiência diagnóstica.
Title in English
Evaluation of glycolipoprotein antigen for serodiagnosis of leptospirosis
Keywords in English
Dot-ELISA
GLP
Glycolipoprotein
Immunodiagnosis
Leptospirosis
Abstract in English
The aim of this work was the study of serologic response against leptospira glycolipoprotein (GLP) for the purpose of standardizing its use as an antigen on serological tests to diagnose human leptospirosis. Among the proteins involved in the pathogenicity, GLP is yet to be studied regarding its immunogenicity and its use as an antigen in the detection of specific antibodies. That led to our standardization of dot-ELISA for detecting IgG and IgM antibodies, using GLP extracted from either pathogenic Leptospira interrogans serovar Copenhageni or nonpathogenic Leptospira biflexa serovar Patoc. Paired serum samples were taken from 90 patients with serologically confirmed leptospirosis, by MAT, with the first samples collected on the disease's acute phase and the second samples after a period of 10 to 15 days. We also tested single samples taken from 30 patients with other diseases, MAT negative, belonging to the control group. Detection rates for IgG antibodies were unsatisfactory, so the study for that use was discontinued. The dot-ELISA IgM results yielded 100% sensitivity on the samples taken after MAT seroconversion. The early detection pattern was evidenced by the high positivity rate on the samples taken during the disease's acute phase (76,6% dot-ELISA using Leptospira interrogans serovar Copenhageni antigen and 90% using Leptospira biflexa sorovar Patoc antigen). Specificity rates were high (96.6%), although a 3.3% rate of false-positive results was observed for dot-ELISA using both antigens. The present study revealed the importance of the humoral immune response against the GLP antigen. The use of GLP, on the dot-ELISA test for IgM antibodies afforded a simple, quick and effective diagnosis.
 
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RobertaMorozetti.pdf (434.20 Kbytes)
Publishing Date
2007-04-25
 
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