Caracteriza????o de isolados brasileiros de Aspergillus terreus por espectrometria de massa MALDI-TOF

Detalhes bibliográficos
Autor(a) principal: Motta, Dielle de Oliveira
Data de Publicação: 2014
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações da UCB
Texto Completo: https://bdtd.ucb.br:8443/jspui/handle/tede/2257
Resumo: The identification of filamentous fungiis traditionally based on morphological and biochemical criteria which are time demanding and for which results can be inaccurate. This fact motivated the evaluation of matrix-assisted laser desorption ionization ??? time of flight (MALDI-TOF) mass spectrometry for the identification of 36 brazilian isolates of Aspergillus terreus. The species is important worldwide due to its ability to produce lovastatin, a drug extremely successful in decreasing plasmatic cholesterol and consequently decreasing the risk of cardiovascular disease, main cause of death worldwide. Prior to MS analysis, the sequencing of internal transcribed spacer (ITS) allowed the identity confirmation of 94% of samples when compared GenBank database. First of all with the aim to compare the molecular biological technique with identification for MALDI-TOF was evaluated the influence of important factors in the quality of mass spectrums and then selected those more suitable to the identification of the specie. This enabled to characterize the proteic profile 30 of the 36 isolates. Nevertheless only 20% of brazilian isolates were identified as A. terreus when compared at the almost 4000 organisms of Biotyper database which may be attributed to the low representation of the specie. The results show that there are strong evidences that the technique is suitable which corroborates with this is: the similar pattern in the dendograms generated for each methodology, the ability to distinguish of the isolates comparatively higher by MALDI-TOF and the proven effectiveness of the technique evaluated by dark tests from the construction of a personal library. For these reasons the use of MALDI-TOF in the identification of isolates of A. terreus in replacing the conventional techniques is promising nevertheless requires optimizations regarding standardizing the processing of samples and inclusion of more reference spectra in the specie in database.
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spelling Magalh??es, Beatriz Simashttp://lattes.cnpq.br/1165242542295220Parachin, N??dia Skorupahttp://lattes.cnpq.br/1214716137780644http://lattes.cnpq.br/9018158431889922Motta, Dielle de Oliveira2017-09-04T12:53:20Z2014-01-01MOTTA, Dielle de Oliveira. Caracteriza????o de isolados brasileiros de Aspergillus terreus por espectrometria de massa MALDI-TOF. 2014. 125 f. Disserta????o (Programa Stricto Sensu em Ci??ncias Gen??micas e Biotecnologia) - Universidade Cat??lica de Bras??lia, Bras??lia, 2014.https://bdtd.ucb.br:8443/jspui/handle/tede/2257The identification of filamentous fungiis traditionally based on morphological and biochemical criteria which are time demanding and for which results can be inaccurate. This fact motivated the evaluation of matrix-assisted laser desorption ionization ??? time of flight (MALDI-TOF) mass spectrometry for the identification of 36 brazilian isolates of Aspergillus terreus. The species is important worldwide due to its ability to produce lovastatin, a drug extremely successful in decreasing plasmatic cholesterol and consequently decreasing the risk of cardiovascular disease, main cause of death worldwide. Prior to MS analysis, the sequencing of internal transcribed spacer (ITS) allowed the identity confirmation of 94% of samples when compared GenBank database. First of all with the aim to compare the molecular biological technique with identification for MALDI-TOF was evaluated the influence of important factors in the quality of mass spectrums and then selected those more suitable to the identification of the specie. This enabled to characterize the proteic profile 30 of the 36 isolates. Nevertheless only 20% of brazilian isolates were identified as A. terreus when compared at the almost 4000 organisms of Biotyper database which may be attributed to the low representation of the specie. The results show that there are strong evidences that the technique is suitable which corroborates with this is: the similar pattern in the dendograms generated for each methodology, the ability to distinguish of the isolates comparatively higher by MALDI-TOF and the proven effectiveness of the technique evaluated by dark tests from the construction of a personal library. For these reasons the use of MALDI-TOF in the identification of isolates of A. terreus in replacing the conventional techniques is promising nevertheless requires optimizations regarding standardizing the processing of samples and inclusion of more reference spectra in the specie in database.A identifica????o de fungos filamentosos ?? tradicionalmente baseada em crit??rios morfol??gicos e bioqu??micos que demandam tempo e cujos resultados podem ser imprecisos. Isso motivou a avalia????o do uso da t??cnica alternativa de espectrometria de massa por ioniza????o e dessor????o a laser assistida por matriz (MALDI-TOF) para a identifica????o de 36 isolados brasileiros de Aspergillus terreus. A esp??cie ?? importante mundialmente, dentre outros motivos, devido ?? habilidade de produzir lovastatina, uma droga bem sucedida na redu????o do colesterol plasm??tico com consequente redu????o do risco de doen??as cardiovasculares, principal causa mundial de ??bitos. Antes das an??lises por MALDI-TOF, o sequenciamento do espa??ador interno transcrito (ITS) permitiu a confirma????o da identidade de 94% (34/36) dos isolados quando comparado ?? base de dados GenBank. Inicialmente de forma a comparar a t??cnica de biologia molecular com a identifica????o por MALDI-TOF foi avaliado a influ??ncia de importantes fatores na qualidade dos dados e selecionado aqueles que mais se adequavam a identifica????o da esp??cie. Isso permitiu caracterizar quanto ao perfil proteico 30 dos 36 isolados. Contudo, somente 20% dos isolados brasileiros foram identificados como A. terreus quando comparado aos quase 4000 organismos da base de dados Biotyper o que pode ser atribu??do ?? baixa representatividade da esp??cie. Segundo os resultados existem fortes evid??ncias de que a t??cnica ?? adequada o que corrobora com isso ?? o padr??o semelhante observado nos dendrogramas gerados por cada uma das metodologias, a capacidade de distin????o dos isolados comparativamente maior por MALDI e a comprovada efic??cia da t??cnica avaliada pelo teste ??s escuras a partir da constru????o de uma biblioteca pr??pria. Por essas raz??es o emprego de MALDI-TOF na identifica????o de isolados de A. terreus em substitui????o as t??cnicas tradicionais ?? promissora, por??m requer otimiza????es no que diz respeito ?? padroniza????o quanto ao processamento das amostras e o incremento de mais espectros de refer??ncia da esp??cie no banco de dados.Submitted by Sara Ribeiro (sara.ribeiro@ucb.br) on 2017-09-04T12:53:05Z No. of bitstreams: 1 DielledeOliveiraMottaDissertacao2014.pdf: 3579770 bytes, checksum: 082642eeca29a044c3ecc5a70434c6bb (MD5)Approved for entry into archive by Sara Ribeiro (sara.ribeiro@ucb.br) on 2017-09-04T12:53:20Z (GMT) No. of bitstreams: 1 DielledeOliveiraMottaDissertacao2014.pdf: 3579770 bytes, checksum: 082642eeca29a044c3ecc5a70434c6bb (MD5)Made available in DSpace on 2017-09-04T12:53:20Z (GMT). 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dc.title.por.fl_str_mv Caracteriza????o de isolados brasileiros de Aspergillus terreus por espectrometria de massa MALDI-TOF
title Caracteriza????o de isolados brasileiros de Aspergillus terreus por espectrometria de massa MALDI-TOF
spellingShingle Caracteriza????o de isolados brasileiros de Aspergillus terreus por espectrometria de massa MALDI-TOF
Motta, Dielle de Oliveira
Identifica????o
Sequenciamento
Aspergillus terreus
MALDI
CNPQ::CIENCIAS DA SAUDE
title_short Caracteriza????o de isolados brasileiros de Aspergillus terreus por espectrometria de massa MALDI-TOF
title_full Caracteriza????o de isolados brasileiros de Aspergillus terreus por espectrometria de massa MALDI-TOF
title_fullStr Caracteriza????o de isolados brasileiros de Aspergillus terreus por espectrometria de massa MALDI-TOF
title_full_unstemmed Caracteriza????o de isolados brasileiros de Aspergillus terreus por espectrometria de massa MALDI-TOF
title_sort Caracteriza????o de isolados brasileiros de Aspergillus terreus por espectrometria de massa MALDI-TOF
author Motta, Dielle de Oliveira
author_facet Motta, Dielle de Oliveira
author_role author
dc.contributor.advisor1.fl_str_mv Magalh??es, Beatriz Simas
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/1165242542295220
dc.contributor.advisor-co1.fl_str_mv Parachin, N??dia Skorupa
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/1214716137780644
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/9018158431889922
dc.contributor.author.fl_str_mv Motta, Dielle de Oliveira
contributor_str_mv Magalh??es, Beatriz Simas
Parachin, N??dia Skorupa
dc.subject.por.fl_str_mv Identifica????o
Sequenciamento
Aspergillus terreus
MALDI
topic Identifica????o
Sequenciamento
Aspergillus terreus
MALDI
CNPQ::CIENCIAS DA SAUDE
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS DA SAUDE
dc.description.abstract.eng.fl_txt_mv The identification of filamentous fungiis traditionally based on morphological and biochemical criteria which are time demanding and for which results can be inaccurate. This fact motivated the evaluation of matrix-assisted laser desorption ionization ??? time of flight (MALDI-TOF) mass spectrometry for the identification of 36 brazilian isolates of Aspergillus terreus. The species is important worldwide due to its ability to produce lovastatin, a drug extremely successful in decreasing plasmatic cholesterol and consequently decreasing the risk of cardiovascular disease, main cause of death worldwide. Prior to MS analysis, the sequencing of internal transcribed spacer (ITS) allowed the identity confirmation of 94% of samples when compared GenBank database. First of all with the aim to compare the molecular biological technique with identification for MALDI-TOF was evaluated the influence of important factors in the quality of mass spectrums and then selected those more suitable to the identification of the specie. This enabled to characterize the proteic profile 30 of the 36 isolates. Nevertheless only 20% of brazilian isolates were identified as A. terreus when compared at the almost 4000 organisms of Biotyper database which may be attributed to the low representation of the specie. The results show that there are strong evidences that the technique is suitable which corroborates with this is: the similar pattern in the dendograms generated for each methodology, the ability to distinguish of the isolates comparatively higher by MALDI-TOF and the proven effectiveness of the technique evaluated by dark tests from the construction of a personal library. For these reasons the use of MALDI-TOF in the identification of isolates of A. terreus in replacing the conventional techniques is promising nevertheless requires optimizations regarding standardizing the processing of samples and inclusion of more reference spectra in the specie in database.
dc.description.abstract.por.fl_txt_mv A identifica????o de fungos filamentosos ?? tradicionalmente baseada em crit??rios morfol??gicos e bioqu??micos que demandam tempo e cujos resultados podem ser imprecisos. Isso motivou a avalia????o do uso da t??cnica alternativa de espectrometria de massa por ioniza????o e dessor????o a laser assistida por matriz (MALDI-TOF) para a identifica????o de 36 isolados brasileiros de Aspergillus terreus. A esp??cie ?? importante mundialmente, dentre outros motivos, devido ?? habilidade de produzir lovastatina, uma droga bem sucedida na redu????o do colesterol plasm??tico com consequente redu????o do risco de doen??as cardiovasculares, principal causa mundial de ??bitos. Antes das an??lises por MALDI-TOF, o sequenciamento do espa??ador interno transcrito (ITS) permitiu a confirma????o da identidade de 94% (34/36) dos isolados quando comparado ?? base de dados GenBank. Inicialmente de forma a comparar a t??cnica de biologia molecular com a identifica????o por MALDI-TOF foi avaliado a influ??ncia de importantes fatores na qualidade dos dados e selecionado aqueles que mais se adequavam a identifica????o da esp??cie. Isso permitiu caracterizar quanto ao perfil proteico 30 dos 36 isolados. Contudo, somente 20% dos isolados brasileiros foram identificados como A. terreus quando comparado aos quase 4000 organismos da base de dados Biotyper o que pode ser atribu??do ?? baixa representatividade da esp??cie. Segundo os resultados existem fortes evid??ncias de que a t??cnica ?? adequada o que corrobora com isso ?? o padr??o semelhante observado nos dendrogramas gerados por cada uma das metodologias, a capacidade de distin????o dos isolados comparativamente maior por MALDI e a comprovada efic??cia da t??cnica avaliada pelo teste ??s escuras a partir da constru????o de uma biblioteca pr??pria. Por essas raz??es o emprego de MALDI-TOF na identifica????o de isolados de A. terreus em substitui????o as t??cnicas tradicionais ?? promissora, por??m requer otimiza????es no que diz respeito ?? padroniza????o quanto ao processamento das amostras e o incremento de mais espectros de refer??ncia da esp??cie no banco de dados.
description The identification of filamentous fungiis traditionally based on morphological and biochemical criteria which are time demanding and for which results can be inaccurate. This fact motivated the evaluation of matrix-assisted laser desorption ionization ??? time of flight (MALDI-TOF) mass spectrometry for the identification of 36 brazilian isolates of Aspergillus terreus. The species is important worldwide due to its ability to produce lovastatin, a drug extremely successful in decreasing plasmatic cholesterol and consequently decreasing the risk of cardiovascular disease, main cause of death worldwide. Prior to MS analysis, the sequencing of internal transcribed spacer (ITS) allowed the identity confirmation of 94% of samples when compared GenBank database. First of all with the aim to compare the molecular biological technique with identification for MALDI-TOF was evaluated the influence of important factors in the quality of mass spectrums and then selected those more suitable to the identification of the specie. This enabled to characterize the proteic profile 30 of the 36 isolates. Nevertheless only 20% of brazilian isolates were identified as A. terreus when compared at the almost 4000 organisms of Biotyper database which may be attributed to the low representation of the specie. The results show that there are strong evidences that the technique is suitable which corroborates with this is: the similar pattern in the dendograms generated for each methodology, the ability to distinguish of the isolates comparatively higher by MALDI-TOF and the proven effectiveness of the technique evaluated by dark tests from the construction of a personal library. For these reasons the use of MALDI-TOF in the identification of isolates of A. terreus in replacing the conventional techniques is promising nevertheless requires optimizations regarding standardizing the processing of samples and inclusion of more reference spectra in the specie in database.
publishDate 2014
dc.date.issued.fl_str_mv 2014-01-01
dc.date.accessioned.fl_str_mv 2017-09-04T12:53:20Z
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dc.identifier.citation.fl_str_mv MOTTA, Dielle de Oliveira. Caracteriza????o de isolados brasileiros de Aspergillus terreus por espectrometria de massa MALDI-TOF. 2014. 125 f. Disserta????o (Programa Stricto Sensu em Ci??ncias Gen??micas e Biotecnologia) - Universidade Cat??lica de Bras??lia, Bras??lia, 2014.
dc.identifier.uri.fl_str_mv https://bdtd.ucb.br:8443/jspui/handle/tede/2257
identifier_str_mv MOTTA, Dielle de Oliveira. Caracteriza????o de isolados brasileiros de Aspergillus terreus por espectrometria de massa MALDI-TOF. 2014. 125 f. Disserta????o (Programa Stricto Sensu em Ci??ncias Gen??micas e Biotecnologia) - Universidade Cat??lica de Bras??lia, Bras??lia, 2014.
url https://bdtd.ucb.br:8443/jspui/handle/tede/2257
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dc.publisher.initials.fl_str_mv UCB
dc.publisher.country.fl_str_mv Brasil
dc.publisher.department.fl_str_mv Escola de Sa??de e Medicina
publisher.none.fl_str_mv Universidade Cat??lica de Bras??lia
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