Comparison of four extraction methods for the detection of hepatitis B virus DNA in dried blood spot samples

Detalhes bibliográficos
Autor(a) principal: Bezerra, Cristiane Sousa
Data de Publicação: 2021
Outros Autores: Portilho, Moyra Machado, Frota, Cristiane Cunha, Villar, Lívia Melo
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
DOI: 10.1002/mbo3.1161
Texto Completo: https://www.arca.fiocruz.br/handle/icict/47877
Resumo: Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil / Instituto Federal de Educação, Ciência e Tecnologia do Ceará. Departamento de Educação. Fortaleza, CE, Brasil.
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spelling Bezerra, Cristiane SousaPortilho, Moyra MachadoFrota, Cristiane CunhaVillar, Lívia Melo2021-06-25T20:02:41Z2021-06-25T20:02:41Z2021BEZERRA, Cristiane Sousa et al. Comparison of four extraction methods for the detection of hepatitis B virus DNA in dried blood spot samples. Microbiology Open, v. 10, e1161, 8 p, 2021.2045-8827https://www.arca.fiocruz.br/handle/icict/4787710.1002/mbo3.1161engWileyDBSextração de DNAHepatite BPCR em tempo realDBSDNA extractionHepatite Breal-time PCRComparison of four extraction methods for the detection of hepatitis B virus DNA in dried blood spot samplesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil / Instituto Federal de Educação, Ciência e Tecnologia do Ceará. Departamento de Educação. Fortaleza, CE, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil / Fundação Oswaldo Cruz. Instituto de Pesquisas Gonçalo Muniz. Salvador, BA, Brasil.Universidade Federal Do Ceará. Faculdade de Medicina. Departamento de Patologia e Medicina Legal. Fortaleza, CE, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil.The dried blood spot (DBS) samples are a useful resource for viral DNA isolation and important in increasing access to HBV diagnosis. However, the choice of the DNA extraction method is crucial for reliable results. We compared the reliability of four DNA extraction methods using DBS samples for the qualitative and quantitative de tection of HBV. A panel of serially diluted HBV DNA in whole blood was spotted onto filter paper (Whatman 903 paper and Whatman FTA cards). Four methods were used to extract DNA: QIAamp® DNA Blood Mini Kit (Qiagen); High Pure Viral Nucleic Acid Kit (Roche); Invisorb Spin Blood Midi Kit (Invitek), and DBS Genomic DNA Isolation Kit (Norgen Biotek). Two qualitative PCRs for the core and surface gene regions of HBV were used, and in-house real-time PCR was also evaluated. It was possible to detect HBV DNA using all extraction and PCR protocols. The lowest limit of detection was found using Whatman 903 paper, Roche extraction, and qualitative PCR (20 copies of HBV DNA per ml) for the surface/polymerase HBV region. In the case of in-house real-time PCR, the lowest limit of detection was found using both Roche and Qiagen assays (estimated 2 × 103 copies per ml). These results suggest the importance of both the extraction method and PCR protocol in detecting HBV DNA in DBS. This study provides insights into the utility of DBS samples in HBV molecular diagnosis and their feasibility in low resource areas where cold storage and transportation may be difficult.info: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/47877/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALCristianeBezerra_LiviaVilar_etal_IOC_2021.pdfCristianeBezerra_LiviaVilar_etal_IOC_2021.pdfapplication/pdf680918https://www.arca.fiocruz.br/bitstream/icict/47877/2/CristianeBezerra_LiviaVilar_etal_IOC_2021.pdffac477dab86638a0c2eb5dc1294c7d8bMD52TEXTCristianeBezerra_LiviaVilar_etal_IOC_2021.pdf.txtCristianeBezerra_LiviaVilar_etal_IOC_2021.pdf.txtExtracted 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dc.title.pt_BR.fl_str_mv Comparison of four extraction methods for the detection of hepatitis B virus DNA in dried blood spot samples
title Comparison of four extraction methods for the detection of hepatitis B virus DNA in dried blood spot samples
spellingShingle Comparison of four extraction methods for the detection of hepatitis B virus DNA in dried blood spot samples
Bezerra, Cristiane Sousa
DBS
extração de DNA
Hepatite B
PCR em tempo real
DBS
DNA extraction
Hepatite B
real-time PCR
title_short Comparison of four extraction methods for the detection of hepatitis B virus DNA in dried blood spot samples
title_full Comparison of four extraction methods for the detection of hepatitis B virus DNA in dried blood spot samples
title_fullStr Comparison of four extraction methods for the detection of hepatitis B virus DNA in dried blood spot samples
title_full_unstemmed Comparison of four extraction methods for the detection of hepatitis B virus DNA in dried blood spot samples
title_sort Comparison of four extraction methods for the detection of hepatitis B virus DNA in dried blood spot samples
author Bezerra, Cristiane Sousa
author_facet Bezerra, Cristiane Sousa
Portilho, Moyra Machado
Frota, Cristiane Cunha
Villar, Lívia Melo
author_role author
author2 Portilho, Moyra Machado
Frota, Cristiane Cunha
Villar, Lívia Melo
author2_role author
author
author
dc.contributor.author.fl_str_mv Bezerra, Cristiane Sousa
Portilho, Moyra Machado
Frota, Cristiane Cunha
Villar, Lívia Melo
dc.subject.other.pt_BR.fl_str_mv DBS
extração de DNA
Hepatite B
PCR em tempo real
topic DBS
extração de DNA
Hepatite B
PCR em tempo real
DBS
DNA extraction
Hepatite B
real-time PCR
dc.subject.en.pt_BR.fl_str_mv DBS
DNA extraction
Hepatite B
real-time PCR
description Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil / Instituto Federal de Educação, Ciência e Tecnologia do Ceará. Departamento de Educação. Fortaleza, CE, Brasil.
publishDate 2021
dc.date.accessioned.fl_str_mv 2021-06-25T20:02:41Z
dc.date.available.fl_str_mv 2021-06-25T20:02:41Z
dc.date.issued.fl_str_mv 2021
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 BEZERRA, Cristiane Sousa et al. Comparison of four extraction methods for the detection of hepatitis B virus DNA in dried blood spot samples. Microbiology Open, v. 10, e1161, 8 p, 2021.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/47877
dc.identifier.issn.pt_BR.fl_str_mv 2045-8827
dc.identifier.doi.none.fl_str_mv 10.1002/mbo3.1161
identifier_str_mv BEZERRA, Cristiane Sousa et al. Comparison of four extraction methods for the detection of hepatitis B virus DNA in dried blood spot samples. Microbiology Open, v. 10, e1161, 8 p, 2021.
2045-8827
10.1002/mbo3.1161
url https://www.arca.fiocruz.br/handle/icict/47877
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
publisher.none.fl_str_mv Wiley
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/47877/1/license.txt
https://www.arca.fiocruz.br/bitstream/icict/47877/2/CristianeBezerra_LiviaVilar_etal_IOC_2021.pdf
https://www.arca.fiocruz.br/bitstream/icict/47877/3/CristianeBezerra_LiviaVilar_etal_IOC_2021.pdf.txt
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repository.name.fl_str_mv Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)
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