Diagnóstico molecular da hanseníase com biomarcadores ML0024 e 85B pela PCR em tempo real
Autor(a) principal: | |
---|---|
Data de Publicação: | 2010 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFU |
Texto Completo: | https://repositorio.ufu.br/handle/123456789/12707 |
Resumo: | Leprosy is a spectral disease caused by Mycobacterium leprae, whose clinical and laboratory diagnosis is performed by means of sputum smear techniques of low sensitivity and specificity, they do not detect the bacilli in paucibacillary (PB). In the present study were tested by qPCR molecular markers, in order to assist the diagnosis of disease, mainly in the form PB. The total DNA of skin lesion biopsy was taken from 185 treatment-naïve patients. The qPCR was performed using primer pairs for amplification of the sequence ML0024 and 85B of the genome of the bacillus. Both markers were highly specific. Detection of DNA of the bacillus by qPCR and qPCR ML0024 85B was 59.45% (110/185) and 57.29% (106/085), respectively, while the bacterial index (BI) smear and dermal skin lesion detected, respectively, the presence of the bacillus in 45.94% (85/185) and 44.86% (83/185) of cases. Among all cases negative for the IB dermal smear, the qPCR and qPCR ML0024 85B detected the presence of the bacillus DNA in 26.00% (26/100) and 23.00% (23/100), respectively. Among those negative for the IB skin lesions, the ML0024 qPCR was positive in 27.45% (28/102) and qPCR 85B in 23.52% (24/102). There was significant difference between positive molecular tests and two tests bacilloscopic (p <0.0001) for the clinical forms of T, and DT-DT +. The agreement between the results of qPCR and qPCR ML0024 85B was more than 90.00% and showed excellent repeatability. Among the tests and molecular tests bacilloscopic the concordance observed were higher than 80.00%. As for the quantitative analysis, we observed a positive correlation between the clinical forms and bacillary load. Molecular markers evaluated in this study also have the same number of copies in the genome, are specific, have virtually the same amplicon size and have similar behavior, although with significant differences in sensitivity. Therefore, this study corroborates with the Ridley- Jopling classification (1966) showing the presence of DNA of M. leprae in samples bacilloscopic whose tests were negative, correlating directly with the bacterial load of each clinical spectrum of disease, increasing certainty in the diagnosis of leprosy. |
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2016-06-22T18:33:03Z2011-10-032016-06-22T18:33:03Z2010-11-17FIGUEIRA, Márcia Moura Nunes Rocha. Diagnóstico molecular da hanseníase com biomarcadores ML0024 e 85B pela PCR em tempo real. 2010. 80 f. Dissertação (Mestrado em Ciências da Saúde) - Universidade Federal de Uberlândia, Uberlândia, 2010.https://repositorio.ufu.br/handle/123456789/12707Leprosy is a spectral disease caused by Mycobacterium leprae, whose clinical and laboratory diagnosis is performed by means of sputum smear techniques of low sensitivity and specificity, they do not detect the bacilli in paucibacillary (PB). In the present study were tested by qPCR molecular markers, in order to assist the diagnosis of disease, mainly in the form PB. The total DNA of skin lesion biopsy was taken from 185 treatment-naïve patients. The qPCR was performed using primer pairs for amplification of the sequence ML0024 and 85B of the genome of the bacillus. Both markers were highly specific. Detection of DNA of the bacillus by qPCR and qPCR ML0024 85B was 59.45% (110/185) and 57.29% (106/085), respectively, while the bacterial index (BI) smear and dermal skin lesion detected, respectively, the presence of the bacillus in 45.94% (85/185) and 44.86% (83/185) of cases. Among all cases negative for the IB dermal smear, the qPCR and qPCR ML0024 85B detected the presence of the bacillus DNA in 26.00% (26/100) and 23.00% (23/100), respectively. Among those negative for the IB skin lesions, the ML0024 qPCR was positive in 27.45% (28/102) and qPCR 85B in 23.52% (24/102). There was significant difference between positive molecular tests and two tests bacilloscopic (p <0.0001) for the clinical forms of T, and DT-DT +. The agreement between the results of qPCR and qPCR ML0024 85B was more than 90.00% and showed excellent repeatability. Among the tests and molecular tests bacilloscopic the concordance observed were higher than 80.00%. As for the quantitative analysis, we observed a positive correlation between the clinical forms and bacillary load. Molecular markers evaluated in this study also have the same number of copies in the genome, are specific, have virtually the same amplicon size and have similar behavior, although with significant differences in sensitivity. Therefore, this study corroborates with the Ridley- Jopling classification (1966) showing the presence of DNA of M. leprae in samples bacilloscopic whose tests were negative, correlating directly with the bacterial load of each clinical spectrum of disease, increasing certainty in the diagnosis of leprosy.Hanseníase é uma doença espectral, causada pelo Mycobacterium leprae, cujo diagnóstico é clínico-laboratorial, realizado por meio de técnicas baciloscópicas de baixa sensibilidade e especificidade, que não detectam o bacilo nas formas paucibacilares (PB). No presente estudo foram testados marcadores moleculares por qPCR, com a finalidade de auxiliar o diagnóstico da doença, principalmente na forma PB. O DNA total de biópsia de lesão de pele foi extraído de 185 pacientes virgens de tratamento. A qPCR foi realizada utilizando pares de primers para amplificação da sequência ML0024 e 85B do genoma do bacilo. Ambos marcadores foram altamente específicos. A detecção do DNA do bacilo pela qPCR ML0024 e qPCR 85B foi 59,45% (110/185) e 57,29% (106/085), respectivamente, enquanto o Índice Baciloscópico (IB) de esfregaço dérmico e de lesão de pele detectaram, respectivamente, a presença do bacilo em 45,94% (85/185) e 44,86% (83/185) dos casos. Dentre todos os casos negativos para o IB de esfregaço dérmico, a qPCR ML0024 e qPCR 85B detectaram a presença do DNA do bacilo em 26,00% (26/100) e 23,00% (23/100), respectivamente. Entre aqueles negativos para o IB de lesão de pele, a qPCR ML0024 foi positiva em 27,45% (28/102) e a qPCR 85B em 23,52% (24/102). Observou-se diferença significativa entre a positividade dos testes moleculares e os dois testes baciloscópicos (p<0,0001) para as formas clínicas T, DT- e DT+. A concordância entre os resultados da qPCR ML0024 e qPCR 85B foi superior a 90,00% e apresentou excelente replicabilidade. Entre os testes baciloscópicos e os testes moleculares as concordâncias observadas foram superiores a 80,00%. Quanto à análise quantitativa, observou-se correlação positiva entre as formas clínicas e a carga bacilar. Os marcadores moleculares avaliados neste estudo possuem igualmente o mesmo número de cópias no genoma, são específicos, apresentam praticamente o mesmo tamanho de amplicon e possuem comportamentos semelhantes, embora não apresentando diferenças significativas na sensibilidade. Portanto, esse trabalho vem corroborar com a classificação de Ridley-Jopling (1966) mostrando a presença de DNA do M. leprae em amostras cujos testes baciloscópicos foram negativos; correlacionando-se diretamente com a carga bacilar de cada forma clínica do espectro da doença, ampliando o diagnóstico de certeza na hanseníase.Fundação de Amparo a Pesquisa do Estado de Minas GeraisMestre em Ciências da Saúdeapplication/pdfporUniversidade Federal de UberlândiaPrograma de Pós-graduação em Ciências da SaúdeUFUBRCiências da SaúdeHanseníaseMycobacterium lepraePelePCR em tempo realPacienteImunohistoquímicaImunologiaPele - DoençasLeprosySkinReal time PCRPatientCNPQ::CIENCIAS DA SAUDEDiagnóstico molecular da hanseníase com biomarcadores ML0024 e 85B pela PCR em tempo realinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisGoulart, Isabela Maria Bernardeshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4703621D8Neves, Adriana Freitashttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4766019A2Vieira, Carlos Ueirahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4706664A7Goulart, Vivian Alonsohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4763812J7http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4713894T8Figueira, Márcia Moura Nunes Rochainfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFUinstname:Universidade Federal de Uberlândia (UFU)instacron:UFUTHUMBNAILDiagnosticoMolecularHanseniase.pdf.jpgDiagnosticoMolecularHanseniase.pdf.jpgGenerated Thumbnailimage/jpeg1180https://repositorio.ufu.br/bitstream/123456789/12707/3/DiagnosticoMolecularHanseniase.pdf.jpge260d034035feaa732718f402039b9c4MD53ORIGINALDiagnosticoMolecularHanseniase.pdfapplication/pdf2718158https://repositorio.ufu.br/bitstream/123456789/12707/1/DiagnosticoMolecularHanseniase.pdf149b83488beb696dabdaa0f187cc3eccMD51TEXTDiagnosticoMolecularHanseniase.pdf.txtDiagnosticoMolecularHanseniase.pdf.txtExtracted texttext/plain100122https://repositorio.ufu.br/bitstream/123456789/12707/2/DiagnosticoMolecularHanseniase.pdf.txtdb3be9fbc1c44a476c2fe17acca0f890MD52123456789/127072016-06-23 03:09:50.537oai:repositorio.ufu.br:123456789/12707Repositório InstitucionalONGhttp://repositorio.ufu.br/oai/requestdiinf@dirbi.ufu.bropendoar:2016-06-23T06:09:50Repositório Institucional da UFU - Universidade Federal de Uberlândia (UFU)false |
dc.title.por.fl_str_mv |
Diagnóstico molecular da hanseníase com biomarcadores ML0024 e 85B pela PCR em tempo real |
title |
Diagnóstico molecular da hanseníase com biomarcadores ML0024 e 85B pela PCR em tempo real |
spellingShingle |
Diagnóstico molecular da hanseníase com biomarcadores ML0024 e 85B pela PCR em tempo real Figueira, Márcia Moura Nunes Rocha Hanseníase Mycobacterium leprae Pele PCR em tempo real Paciente Imunohistoquímica Imunologia Pele - Doenças Leprosy Skin Real time PCR Patient CNPQ::CIENCIAS DA SAUDE |
title_short |
Diagnóstico molecular da hanseníase com biomarcadores ML0024 e 85B pela PCR em tempo real |
title_full |
Diagnóstico molecular da hanseníase com biomarcadores ML0024 e 85B pela PCR em tempo real |
title_fullStr |
Diagnóstico molecular da hanseníase com biomarcadores ML0024 e 85B pela PCR em tempo real |
title_full_unstemmed |
Diagnóstico molecular da hanseníase com biomarcadores ML0024 e 85B pela PCR em tempo real |
title_sort |
Diagnóstico molecular da hanseníase com biomarcadores ML0024 e 85B pela PCR em tempo real |
author |
Figueira, Márcia Moura Nunes Rocha |
author_facet |
Figueira, Márcia Moura Nunes Rocha |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Goulart, Isabela Maria Bernardes |
dc.contributor.advisor1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4703621D8 |
dc.contributor.referee1.fl_str_mv |
Neves, Adriana Freitas |
dc.contributor.referee1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4766019A2 |
dc.contributor.referee2.fl_str_mv |
Vieira, Carlos Ueira |
dc.contributor.referee2Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4706664A7 |
dc.contributor.referee3.fl_str_mv |
Goulart, Vivian Alonso |
dc.contributor.referee3Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4763812J7 |
dc.contributor.authorLattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4713894T8 |
dc.contributor.author.fl_str_mv |
Figueira, Márcia Moura Nunes Rocha |
contributor_str_mv |
Goulart, Isabela Maria Bernardes Neves, Adriana Freitas Vieira, Carlos Ueira Goulart, Vivian Alonso |
dc.subject.por.fl_str_mv |
Hanseníase Mycobacterium leprae Pele PCR em tempo real Paciente Imunohistoquímica Imunologia Pele - Doenças |
topic |
Hanseníase Mycobacterium leprae Pele PCR em tempo real Paciente Imunohistoquímica Imunologia Pele - Doenças Leprosy Skin Real time PCR Patient CNPQ::CIENCIAS DA SAUDE |
dc.subject.eng.fl_str_mv |
Leprosy Skin Real time PCR Patient |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS DA SAUDE |
description |
Leprosy is a spectral disease caused by Mycobacterium leprae, whose clinical and laboratory diagnosis is performed by means of sputum smear techniques of low sensitivity and specificity, they do not detect the bacilli in paucibacillary (PB). In the present study were tested by qPCR molecular markers, in order to assist the diagnosis of disease, mainly in the form PB. The total DNA of skin lesion biopsy was taken from 185 treatment-naïve patients. The qPCR was performed using primer pairs for amplification of the sequence ML0024 and 85B of the genome of the bacillus. Both markers were highly specific. Detection of DNA of the bacillus by qPCR and qPCR ML0024 85B was 59.45% (110/185) and 57.29% (106/085), respectively, while the bacterial index (BI) smear and dermal skin lesion detected, respectively, the presence of the bacillus in 45.94% (85/185) and 44.86% (83/185) of cases. Among all cases negative for the IB dermal smear, the qPCR and qPCR ML0024 85B detected the presence of the bacillus DNA in 26.00% (26/100) and 23.00% (23/100), respectively. Among those negative for the IB skin lesions, the ML0024 qPCR was positive in 27.45% (28/102) and qPCR 85B in 23.52% (24/102). There was significant difference between positive molecular tests and two tests bacilloscopic (p <0.0001) for the clinical forms of T, and DT-DT +. The agreement between the results of qPCR and qPCR ML0024 85B was more than 90.00% and showed excellent repeatability. Among the tests and molecular tests bacilloscopic the concordance observed were higher than 80.00%. As for the quantitative analysis, we observed a positive correlation between the clinical forms and bacillary load. Molecular markers evaluated in this study also have the same number of copies in the genome, are specific, have virtually the same amplicon size and have similar behavior, although with significant differences in sensitivity. Therefore, this study corroborates with the Ridley- Jopling classification (1966) showing the presence of DNA of M. leprae in samples bacilloscopic whose tests were negative, correlating directly with the bacterial load of each clinical spectrum of disease, increasing certainty in the diagnosis of leprosy. |
publishDate |
2010 |
dc.date.issued.fl_str_mv |
2010-11-17 |
dc.date.available.fl_str_mv |
2011-10-03 2016-06-22T18:33:03Z |
dc.date.accessioned.fl_str_mv |
2016-06-22T18:33:03Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
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masterThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
FIGUEIRA, Márcia Moura Nunes Rocha. Diagnóstico molecular da hanseníase com biomarcadores ML0024 e 85B pela PCR em tempo real. 2010. 80 f. Dissertação (Mestrado em Ciências da Saúde) - Universidade Federal de Uberlândia, Uberlândia, 2010. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufu.br/handle/123456789/12707 |
identifier_str_mv |
FIGUEIRA, Márcia Moura Nunes Rocha. Diagnóstico molecular da hanseníase com biomarcadores ML0024 e 85B pela PCR em tempo real. 2010. 80 f. Dissertação (Mestrado em Ciências da Saúde) - Universidade Federal de Uberlândia, Uberlândia, 2010. |
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https://repositorio.ufu.br/handle/123456789/12707 |
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por |
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Universidade Federal de Uberlândia |
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Programa de Pós-graduação em Ciências da Saúde |
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UFU |
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BR |
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Ciências da Saúde |
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Universidade Federal de Uberlândia |
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UFU |
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Repositório Institucional da UFU |
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