A rapid and accurate methylation-sensitive high-resolution melting analysis assay for the diagnosis of Prader Willi and Angelman patients

Detalhes bibliográficos
Autor(a) principal: Ferreira, Igor Ribeiro
Data de Publicação: 2019
Outros Autores: Cunha, Wilton Darleans dos Santos, Gomes, Leonardo Henrique Ferreira, Cintra, Hiago Azevedo, Fonseca, Letícia Lopes Cabral Guimarães, Ferreira Bastos, Elenice Ferreira, Llerena Junior, Juan Clinton, Vasconcelos, Zilton Farias Meira de, Guida, Letícia da Cunha
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/42141
Resumo: This work was supported by “Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPQ)” and was financed in part by the “Coordenação de Aperfeiçoamento de Pessoal de Nível Superior ‐ Brasil (CAPES) ‐ Finance Code 001”. All the experiments were developed in the technological platforms “RPT01A ‐ Sequencing of DNA”, “RPT09G ‐ and RPT09I ‐ PCR in Real Time – IFF”, belonging to FIOCRUZ Network of Technical. Platforms (PDTIS/FIOCRUZ).
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spelling Ferreira, Igor RibeiroCunha, Wilton Darleans dos SantosGomes, Leonardo Henrique FerreiraCintra, Hiago AzevedoFonseca, Letícia Lopes Cabral GuimarãesFerreira Bastos, Elenice FerreiraLlerena Junior, Juan ClintonVasconcelos, Zilton Farias Meira deGuida, Letícia da Cunha2020-07-07T17:37:13Z2020-07-07T17:37:13Z2019FERREIRA, Igor Ribeiro et al.A rapid and accurate methylation‐sensitive high‐resolution melting analysis assay for the diagnosis of Prader Willi and Angelman patients. Molecular Genetics & Genomic Medicine, v. 7, p. 1-10, e637, 2019.2324-9269https://www.arca.fiocruz.br/handle/icict/4214110.1002/mgg3.637This work was supported by “Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPQ)” and was financed in part by the “Coordenação de Aperfeiçoamento de Pessoal de Nível Superior ‐ Brasil (CAPES) ‐ Finance Code 001”. All the experiments were developed in the technological platforms “RPT01A ‐ Sequencing of DNA”, “RPT09G ‐ and RPT09I ‐ PCR in Real Time – IFF”, belonging to FIOCRUZ Network of Technical. Platforms (PDTIS/FIOCRUZ).Fundo: As síndromes de Prader Willi (PWS) e Angelman (AS) são raros distúrbios genéticos caracterizados por deleções, disomias uniparentais e defeitos de impressão no cromossomo 15. A perda de função de genes específicos causados ​​por alterações genéticas no alelo paterno causa SPW enquanto a ausência na mãe resultados do alelo AS. O diagnóstico laboratorial de SPW e SA é complexo e exige técnicas de biologia molecular e citogenética para identificar o mecanismo genético relacionado ao desenvolvimento da doença. A análise de metilação do DNA no cromossomo 15 no locus SNURF ‐ SNRPN através do MS ‐ PCR confirma o diagnóstico e distingue entre PWS e AS. Nosso estudo teve como objetivo estabelecer a técnica MS ‐ PCR associada ao derretimento de alta resolução (MS ‐ HRM) no diagnóstico de SPW e SA com um único par de primers. Métodos: Coletamos amostras de sangue de 43 pacientes suspeitos para uma análise citogenética e de metilação. O DNA extraído foi tratado com bissulfito para realizar análises comparativas de metilação. Resultados: MS ‐ HRM e MS ‐ PCR concordaram em 100% dos casos, identificando 19 (44%) SPW, 3 (7%) AS e 21 (49%) Normal. A análise FISH detectou quatro casos de SPW causados ​​por deleções no cromossomo 15. Conclusão: O MS ‐ HRM apresentou bom desempenho com um par exclusivo de primers, dispensando a análise de gel de eletroforese, oferecendo um diagnóstico rápido e reprodutível.Background: Prader Willi (PWS) and Angelman (AS) syndromes are rare genetic disorders characterized by deletions, uniparental disomy, and imprinting defects at chromosome 15. The loss of function of specific genes caused by genetic alterations in paternal allele causes PWS while the absence in maternal allele results AS. The laboratory diagnosis of PWS and AS is complex and demands molecular biology and cytogenetics techniques to identify the genetic mechanism related to the development of the disease. The DNA methylation analysis in chromosome 15 at the SNURF‐SNRPN locus through MS‐PCR confirms the diagnosis and distinguishes between PWS and AS. Our study aimed to establish the MS‐PCR technique associated with High‐Resolution Melting (MS‐HRM) in PWS and AS diagnostic with a single pair of primers. Methods: We collected blood samples from 43 suspected patients to a cytogenetic and methylation analysis. The extracted DNA was treated with bisulfite to perform comparative methylation analysis. Results: MS‐HRM and MS‐PCR agreed in 100% of cases, identifying 19(44%) PWS, 3(7%) AS, and 21(49%) Normal. FISH analysis detected four cases of PWS caused by deletions in chromosome 15. Conclusion: The MS‐HRM showed good performance with a unique pair of primers, dispensing electrophoresis gel analysis, offering a quick and reproducible diagnostic.Conselho Nacional de Desenvolvimento Científico e Tecnológico, CNPQ (Pesquisa sobre Doenças Raras – 35/2014); Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, CAPES.Fundação Oswaldo Cruz. Instituto Nacional da Saúde da Mulher da Criança e do Adolescente Fernandes Figueira. Laboratório de Alta Complexidade. Rio de Janeiro, RJ. Brasil,Fundação Oswaldo Cruz. Instituto Nacional da Saúde da Mulher da Criança e do Adolescente Fernandes Figueira. Laboratório de Alta Complexidade. Rio de Janeiro, RJ. Brasil,Fundação Oswaldo Cruz. Instituto Nacional da Saúde da Mulher da Criança e do Adolescente Fernandes Figueira. Laboratório de Alta Complexidade. Rio de Janeiro, RJ. Brasil,Fundação Oswaldo Cruz. Instituto Nacional da Saúde da Mulher da Criança e do Adolescente Fernandes Figueira. Laboratório de Alta Complexidade. Rio de Janeiro, RJ. Brasil,Fundação Oswaldo Cruz. Instituto Nacional da Saúde da Mulher da Criança e do Adolescente Fernandes Figueira. Departamento de Genética. Rio de Janeiro, RJ. Brasil,Fundação Oswaldo Cruz. Instituto Nacional da Saúde da Mulher da Criança e do Adolescente Fernandes Figueira. Departamento de Genética. Rio de Janeiro, RJ. Brasil,Fundação Oswaldo Cruz. Instituto Nacional da Saúde da Mulher da Criança e do Adolescente Fernandes Figueira. Laboratório de Alta Complexidade. Rio de Janeiro, RJ. Brasil,Fundação Oswaldo Cruz. Instituto Nacional da Saúde da Mulher da Criança e do Adolescente Fernandes Figueira. Laboratório de Alta Complexidade. Rio de Janeiro, RJ. Brasil,engWiley Open AccessSíndrome de Prader WilliMS ‐ PCRMS ‐ HRMFusão de alta resoluçãoSíndrome de AngelmanPrader Willi syndromeMS‐PCRMS‐HRMHigh‐resolution meltingAngelman syndromeA rapid and accurate methylation-sensitive high-resolution melting analysis assay for the diagnosis of Prader Willi and Angelman patientsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-82991https://www.arca.fiocruz.br/bitstream/icict/42141/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALMGG3-7-e637.pdfMGG3-7-e637.pdfapplication/pdf668281https://www.arca.fiocruz.br/bitstream/icict/42141/2/MGG3-7-e637.pdf5bcadab7e0063882f9b0f93eee625bf7MD52TEXTMGG3-7-e637.pdf.txtMGG3-7-e637.pdf.txtExtracted 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dc.title.pt_BR.fl_str_mv A rapid and accurate methylation-sensitive high-resolution melting analysis assay for the diagnosis of Prader Willi and Angelman patients
title A rapid and accurate methylation-sensitive high-resolution melting analysis assay for the diagnosis of Prader Willi and Angelman patients
spellingShingle A rapid and accurate methylation-sensitive high-resolution melting analysis assay for the diagnosis of Prader Willi and Angelman patients
Ferreira, Igor Ribeiro
Síndrome de Prader Willi
MS ‐ PCR
MS ‐ HRM
Fusão de alta resolução
Síndrome de Angelman
Prader Willi syndrome
MS‐PCR
MS‐HRM
High‐resolution melting
Angelman syndrome
title_short A rapid and accurate methylation-sensitive high-resolution melting analysis assay for the diagnosis of Prader Willi and Angelman patients
title_full A rapid and accurate methylation-sensitive high-resolution melting analysis assay for the diagnosis of Prader Willi and Angelman patients
title_fullStr A rapid and accurate methylation-sensitive high-resolution melting analysis assay for the diagnosis of Prader Willi and Angelman patients
title_full_unstemmed A rapid and accurate methylation-sensitive high-resolution melting analysis assay for the diagnosis of Prader Willi and Angelman patients
title_sort A rapid and accurate methylation-sensitive high-resolution melting analysis assay for the diagnosis of Prader Willi and Angelman patients
author Ferreira, Igor Ribeiro
author_facet Ferreira, Igor Ribeiro
Cunha, Wilton Darleans dos Santos
Gomes, Leonardo Henrique Ferreira
Cintra, Hiago Azevedo
Fonseca, Letícia Lopes Cabral Guimarães
Ferreira Bastos, Elenice Ferreira
Llerena Junior, Juan Clinton
Vasconcelos, Zilton Farias Meira de
Guida, Letícia da Cunha
author_role author
author2 Cunha, Wilton Darleans dos Santos
Gomes, Leonardo Henrique Ferreira
Cintra, Hiago Azevedo
Fonseca, Letícia Lopes Cabral Guimarães
Ferreira Bastos, Elenice Ferreira
Llerena Junior, Juan Clinton
Vasconcelos, Zilton Farias Meira de
Guida, Letícia da Cunha
author2_role author
author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Ferreira, Igor Ribeiro
Cunha, Wilton Darleans dos Santos
Gomes, Leonardo Henrique Ferreira
Cintra, Hiago Azevedo
Fonseca, Letícia Lopes Cabral Guimarães
Ferreira Bastos, Elenice Ferreira
Llerena Junior, Juan Clinton
Vasconcelos, Zilton Farias Meira de
Guida, Letícia da Cunha
dc.subject.other.pt_BR.fl_str_mv Síndrome de Prader Willi
MS ‐ PCR
MS ‐ HRM
Fusão de alta resolução
Síndrome de Angelman
topic Síndrome de Prader Willi
MS ‐ PCR
MS ‐ HRM
Fusão de alta resolução
Síndrome de Angelman
Prader Willi syndrome
MS‐PCR
MS‐HRM
High‐resolution melting
Angelman syndrome
dc.subject.en.pt_BR.fl_str_mv Prader Willi syndrome
MS‐PCR
MS‐HRM
High‐resolution melting
Angelman syndrome
description This work was supported by “Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPQ)” and was financed in part by the “Coordenação de Aperfeiçoamento de Pessoal de Nível Superior ‐ Brasil (CAPES) ‐ Finance Code 001”. All the experiments were developed in the technological platforms “RPT01A ‐ Sequencing of DNA”, “RPT09G ‐ and RPT09I ‐ PCR in Real Time – IFF”, belonging to FIOCRUZ Network of Technical. Platforms (PDTIS/FIOCRUZ).
publishDate 2019
dc.date.issued.fl_str_mv 2019
dc.date.accessioned.fl_str_mv 2020-07-07T17:37:13Z
dc.date.available.fl_str_mv 2020-07-07T17:37:13Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.citation.fl_str_mv FERREIRA, Igor Ribeiro et al.A rapid and accurate methylation‐sensitive high‐resolution melting analysis assay for the diagnosis of Prader Willi and Angelman patients. Molecular Genetics & Genomic Medicine, v. 7, p. 1-10, e637, 2019.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/42141
dc.identifier.issn.pt_BR.fl_str_mv 2324-9269
dc.identifier.doi.none.fl_str_mv 10.1002/mgg3.637
identifier_str_mv FERREIRA, Igor Ribeiro et al.A rapid and accurate methylation‐sensitive high‐resolution melting analysis assay for the diagnosis of Prader Willi and Angelman patients. Molecular Genetics & Genomic Medicine, v. 7, p. 1-10, e637, 2019.
2324-9269
10.1002/mgg3.637
url https://www.arca.fiocruz.br/handle/icict/42141
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Wiley Open Access
publisher.none.fl_str_mv Wiley Open Access
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
collection Repositório Institucional da FIOCRUZ (ARCA)
bitstream.url.fl_str_mv https://www.arca.fiocruz.br/bitstream/icict/42141/1/license.txt
https://www.arca.fiocruz.br/bitstream/icict/42141/2/MGG3-7-e637.pdf
https://www.arca.fiocruz.br/bitstream/icict/42141/3/MGG3-7-e637.pdf.txt
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