Comparison of Four Extraction Methods to Detect Hepatitis A Virus RNA in Serum and Stool Samples

Detalhes bibliográficos
Autor(a) principal: Paula, Vanessa Salete de
Data de Publicação: 2003
Outros Autores: Villar, Livia Melo, Gaspar, Ana Maria Coimbra
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/33506
Resumo: Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Virologia. Rio de Janeiro, RJ. Brasil.
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spelling Paula, Vanessa Salete deVillar, Livia MeloGaspar, Ana Maria Coimbra2019-06-13T17:58:05Z2019-06-13T17:58:05Z2003PAULA, Vanessa Salete de; VILLAR, Livia Melo; GASPAR, Ana Maria Coimbra. Comparison of Four Extraction Methods to Detect Hepatitis A Virus RNA in Serum and Stool Samples. The Brazilian Journal of Infectious Diseases, v. 7, n. 2, p. 135-141, 2003.1413-8670https://www.arca.fiocruz.br/handle/icict/3350610.1590/S1413-867020030002000071678-4391engElsevierReação em Cadeia da Polimerase em Tempo RealAnticorpos Anti-Hepatite ASoroHAVExtraction methodsSerum and stoolRT-PCRMatéria fecalComparison of Four Extraction Methods to Detect Hepatitis A Virus RNA in Serum and Stool Samplesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Virologia. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Virologia. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Virologia. Rio de Janeiro, RJ. Brasil.The efficiency of extraction methods for hepatitis A virus (HAV) RNA in clinical samples is of great importance for molecular diagnosis, especially in regions endemic for HAV, such as Brazil. We compared the efficiency of four different extraction techniques in serum and stool samples for the detection of hepatitis A virus by reverse transcription PCR (RT-PCR). We used PCR to analyse serum and stool samples of 12 patients who were referred to the Brazilian Reference Center for Viral Hepatitis (BRCVH) in Rio de Janeiro. The methods tested were Proteinase K, Silica, TRIzol and Guanidine isothiocyanate. Proteinase K extraction was the best method for serum samples; it detected the HAV-RNA in 11 of the 12 samples. The guanidine isothiocyanate method was the most effective for stool samples, detecting HAV-RNAs in 9 of the samples. The TRIzolâ method worked best with serum samples, and the silica method was unsatisfactory with both serum and stool samples. The RNA extraction method affected the outcome. The use of appropriate RNA extraction methods is a critical step for successful and valid PCR studies on clinical samples. We recommend that RNA extraction techniques be carefully selected for their efficiency with each type of specimen.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/33506/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALAnaGaspar_VanessaPaula_IOC_2003.pdfAnaGaspar_VanessaPaula_IOC_2003.pdfapplication/pdf44667https://www.arca.fiocruz.br/bitstream/icict/33506/2/AnaGaspar_VanessaPaula_IOC_2003.pdf8dffe2332199668aa3a3d6911e7eaa01MD52TEXTAnaGaspar_VanessaPaula_IOC_2003.pdf.txtAnaGaspar_VanessaPaula_IOC_2003.pdf.txtExtracted texttext/plain24359https://www.arca.fiocruz.br/bitstream/icict/33506/3/AnaGaspar_VanessaPaula_IOC_2003.pdf.txt7c1bdcf2493d989598641c61b60366d0MD53icict/335062019-06-15 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dc.title.pt_BR.fl_str_mv Comparison of Four Extraction Methods to Detect Hepatitis A Virus RNA in Serum and Stool Samples
title Comparison of Four Extraction Methods to Detect Hepatitis A Virus RNA in Serum and Stool Samples
spellingShingle Comparison of Four Extraction Methods to Detect Hepatitis A Virus RNA in Serum and Stool Samples
Paula, Vanessa Salete de
Reação em Cadeia da Polimerase em Tempo Real
Anticorpos Anti-Hepatite A
Soro
HAV
Extraction methods
Serum and stool
RT-PCR
Matéria fecal
title_short Comparison of Four Extraction Methods to Detect Hepatitis A Virus RNA in Serum and Stool Samples
title_full Comparison of Four Extraction Methods to Detect Hepatitis A Virus RNA in Serum and Stool Samples
title_fullStr Comparison of Four Extraction Methods to Detect Hepatitis A Virus RNA in Serum and Stool Samples
title_full_unstemmed Comparison of Four Extraction Methods to Detect Hepatitis A Virus RNA in Serum and Stool Samples
title_sort Comparison of Four Extraction Methods to Detect Hepatitis A Virus RNA in Serum and Stool Samples
author Paula, Vanessa Salete de
author_facet Paula, Vanessa Salete de
Villar, Livia Melo
Gaspar, Ana Maria Coimbra
author_role author
author2 Villar, Livia Melo
Gaspar, Ana Maria Coimbra
author2_role author
author
dc.contributor.author.fl_str_mv Paula, Vanessa Salete de
Villar, Livia Melo
Gaspar, Ana Maria Coimbra
dc.subject.other.pt_BR.fl_str_mv Reação em Cadeia da Polimerase em Tempo Real
Anticorpos Anti-Hepatite A
Soro
topic Reação em Cadeia da Polimerase em Tempo Real
Anticorpos Anti-Hepatite A
Soro
HAV
Extraction methods
Serum and stool
RT-PCR
Matéria fecal
dc.subject.en.pt_BR.fl_str_mv HAV
Extraction methods
Serum and stool
RT-PCR
dc.subject.decs.pt_BR.fl_str_mv Matéria fecal
description Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Virologia. Rio de Janeiro, RJ. Brasil.
publishDate 2003
dc.date.issued.fl_str_mv 2003
dc.date.accessioned.fl_str_mv 2019-06-13T17:58:05Z
dc.date.available.fl_str_mv 2019-06-13T17:58:05Z
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 PAULA, Vanessa Salete de; VILLAR, Livia Melo; GASPAR, Ana Maria Coimbra. Comparison of Four Extraction Methods to Detect Hepatitis A Virus RNA in Serum and Stool Samples. The Brazilian Journal of Infectious Diseases, v. 7, n. 2, p. 135-141, 2003.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/33506
dc.identifier.issn.pt_BR.fl_str_mv 1413-8670
dc.identifier.doi.pt_BR.fl_str_mv 10.1590/S1413-86702003000200007
dc.identifier.eissn.none.fl_str_mv 1678-4391
identifier_str_mv PAULA, Vanessa Salete de; VILLAR, Livia Melo; GASPAR, Ana Maria Coimbra. Comparison of Four Extraction Methods to Detect Hepatitis A Virus RNA in Serum and Stool Samples. The Brazilian Journal of Infectious Diseases, v. 7, n. 2, p. 135-141, 2003.
1413-8670
10.1590/S1413-86702003000200007
1678-4391
url https://www.arca.fiocruz.br/handle/icict/33506
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 Elsevier
publisher.none.fl_str_mv Elsevier
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)
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