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Doctoral Thesis
DOI
10.11606/T.5.2012.tde-22012013-175521
Document
Author
Full name
Janaina Miranda Bezerra
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2012
Supervisor
Committee
Ferreira, Antonio Walter (President)
Kanunfre, Kelly Aparecida
Granato, Celso Francisco Hernandes
Okay, Thelma Suely
Sato, Maria Notomi
Title in Portuguese
Desempenho de antígenos nativo, recombinante e sintético, em testes imunoenzimáticos, para diagnóstico e prognóstico de pacientes com diferentes formas clínicas de hanseníase
Keywords in Portuguese
Diagnóstico
ELISA
Hanseníase
Mycobacterium leprae
Prognóstico
Western blotting
Abstract in Portuguese
Apesar da Organização Mundial de Saúde (OMS) ter adotado medidas de controle, a hanseníase permanece como um problema social e de saúde pública em muitos países. O diagnóstico clínico e a baciloscopia permanecem como as principais ferramentas utilizadas para a classificação dos pacientes e definição da terapêutica a ser instituída. A identificação de novos antígenos de M. leprae, o desenvolvimento de novos métodos laboratoriais e melhorar o desempenho de testes sorológicos já existentes são prioridades para tentar atingir as metas da OMS programadas para o período de 2011-2015. Neste estudo avaliamos o desempenho diagnóstico dos antígenos glicolipídeo sintético ND-O-BSA para pesquisa de anticorpos totais e recombinante LID-1, para pesquisa anticorpos IgG por ELISA; e o perfil de reatividade de anticorpos IgG contra proteínas nativas de M. leprae por Western Blotting, em amostras de soros de pacientes de área endêmica com diferentes formas clínicas da doença. Nossos resultados mostraram que os testes ELISA ND-O-BSA e LID-1 auxiliaram na detecção de 70% dos pacientes multibacilares com baciloscopia negativa. O valor preditivo negativo foi de 94% para ambos os testes. A análise do Western Blotting mostrou que pacientes multibacilares possuem anticorpos IgG para a fração de 38kDa e região de 3,5kDa, esta reatividade não foi observada no grupo controle. Nosso estudo sugere a utilização do ELISA, com antígenos recombinante e sintético, na rotina diagnóstica; e que a fração de 38kDa e região de 3,5kDa, de antígeno nativo de M. leprae, são bons marcadores no diagnóstico de hanseníase e no prognóstico de pacientes com as formas borderline e indeterminada da doença.
Title in English
Performance of native, recombinant and synthetic antigens by enzyme immunoassays for the diagnosis and prognosis of leprosy patients classified in the wide spectrum of the disease
Keywords in English
Diagnosis
ELISA
Leprosy
Mycobacterium leprae
Prognosis
Western blotting
Abstract in English
Although several control measures have been adopted by the World Health Organization, (WHO) leprosy continues to be a social and public health problem in many countries. Clinical diagnosis and acid-fast bacilli skin smear are the main tools used to classify patients and define therapy. According to the WHO program for 2011-2015, the identification of new Mycobacterium leprae antigens, the development of new laboratory methods and improved serological tests are the priorities. In the present study, we evaluated the diagnostic performance of the synthetic glycolipid antigen, ND-O-BSA, to detect total antibodies and the recombinant antigen LID-1 to detect IgG antibodies by ELISA and the IgG reactivity profile against native M. leprae proteins by Western blot in serum samples from leprosy patients classified in the wide spectrum of the disease. Our results showed that ND-O-BSA and LID-1 ELISA are able to detect the disease in 70% of multibacillary patients with negative skin smears. The negative predictive values were 94% for both tests. The Western blot analysis revealed that most multibacillary patients had IgG antibodies against the 38 kDa and 3.5 kDa regions; this reactivity was not observed in the control group. Our study suggests the use of ELISA, with recombinant or synthetic antigens, in routine diagnosis and that the 38 kDa fraction and the 3.5 kDa region, from native M leprae antigen, are good markers for the diagnosis of leprosy and its prognosis as shown for the borderline and indeterminate forms of the disease.
 
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Publishing Date
2013-01-23
 
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