MELTING CURVE ANALYSIS FOR THE SCREENING OF HEPATITIS B VIRUS GENOTYPES A, D AND F IN PATIENTS FROM A GENERAL HOSPITAL IN SOUTHERN BRAZIL
Autor(a) principal: | |
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Data de Publicação: | 2013 |
Outros Autores: | , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Arquivos de gastroenterologia (Online) |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0004-28032013000300219 |
Resumo: | Context Hepatitis B virus (HBV) can cause fulminant hepatitis, cirrhosis and hepatocellular carcinoma, and is one of the most common causes of acute and chronic liver failure. The genetic variants of HBV can be decisive for the evolution of these diseases as well as for the election of therapy. Objectives The aim of this study was to evaluate and standardize an in house methodology based on the analysis of the melting curve polymerase chain reaction (PCR) of real-time (qPCR) to screen for genotypes A, D and F of HBV in patients from a hospital in Rio Grande do Sul, Brazil. Methods We evaluated 104 patients presumably with HBV chronic infection. Viral DNA was extracted from plasma and viral genotypes and different mutations were determined using PCR-based protocols. Results A PCR-based methodology was standardized for the analysis of genotypes A, D and F of HBV. The technique was based in a nested PCR with the final step consisting of a multiplex real-time PCR, using the melting curve as a tool for the differentiation of fragments. A higher frequency of genotype D (44.4%), followed by genotype A (22.2%) and genotype F (3.7%) was observed. Conclusion The standardized assay, a nested PCR-multiplex qPCR using specific primers, provides a rapid and accurate method for the differentiation of HBV genotypes that are more frequent in Southern Brazil – A, D and F. This method can be applied in the clinical practice. |
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MELTING CURVE ANALYSIS FOR THE SCREENING OF HEPATITIS B VIRUS GENOTYPES A, D AND F IN PATIENTS FROM A GENERAL HOSPITAL IN SOUTHERN BRAZILHepatitis B virusMelting Curve AnalysisGenotypeReal-time polymerase chain reactionMinas Gerais, Brazil Context Hepatitis B virus (HBV) can cause fulminant hepatitis, cirrhosis and hepatocellular carcinoma, and is one of the most common causes of acute and chronic liver failure. The genetic variants of HBV can be decisive for the evolution of these diseases as well as for the election of therapy. Objectives The aim of this study was to evaluate and standardize an in house methodology based on the analysis of the melting curve polymerase chain reaction (PCR) of real-time (qPCR) to screen for genotypes A, D and F of HBV in patients from a hospital in Rio Grande do Sul, Brazil. Methods We evaluated 104 patients presumably with HBV chronic infection. Viral DNA was extracted from plasma and viral genotypes and different mutations were determined using PCR-based protocols. Results A PCR-based methodology was standardized for the analysis of genotypes A, D and F of HBV. The technique was based in a nested PCR with the final step consisting of a multiplex real-time PCR, using the melting curve as a tool for the differentiation of fragments. A higher frequency of genotype D (44.4%), followed by genotype A (22.2%) and genotype F (3.7%) was observed. Conclusion The standardized assay, a nested PCR-multiplex qPCR using specific primers, provides a rapid and accurate method for the differentiation of HBV genotypes that are more frequent in Southern Brazil – A, D and F. This method can be applied in the clinical practice. Instituto Brasileiro de Estudos e Pesquisas de Gastroenterologia e Outras Especialidades - IBEPEGE. 2013-09-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0004-28032013000300219Arquivos de Gastroenterologia v.50 n.3 2013reponame:Arquivos de gastroenterologia (Online)instname:Instituto Brasileiro de Estudos e Pesquisas de Gastroenterologiainstacron:IBEPEGE10.1590/S0004-28032013000200039info:eu-repo/semantics/openAccessBECKER,Carlos EduardoKRETZMANN,Nelson AlexandreMATTOS,Angelo Alves deVEIGA,Ana Beatriz Gorini daeng2013-10-08T00:00:00Zoai:scielo:S0004-28032013000300219Revistahttp://www.scielo.br/aghttps://old.scielo.br/oai/scielo-oai.php||secretariaarqgastr@hospitaligesp.com.br1678-42190004-2803opendoar:2013-10-08T00:00Arquivos de gastroenterologia (Online) - Instituto Brasileiro de Estudos e Pesquisas de Gastroenterologiafalse |
dc.title.none.fl_str_mv |
MELTING CURVE ANALYSIS FOR THE SCREENING OF HEPATITIS B VIRUS GENOTYPES A, D AND F IN PATIENTS FROM A GENERAL HOSPITAL IN SOUTHERN BRAZIL |
title |
MELTING CURVE ANALYSIS FOR THE SCREENING OF HEPATITIS B VIRUS GENOTYPES A, D AND F IN PATIENTS FROM A GENERAL HOSPITAL IN SOUTHERN BRAZIL |
spellingShingle |
MELTING CURVE ANALYSIS FOR THE SCREENING OF HEPATITIS B VIRUS GENOTYPES A, D AND F IN PATIENTS FROM A GENERAL HOSPITAL IN SOUTHERN BRAZIL BECKER,Carlos Eduardo Hepatitis B virus Melting Curve Analysis Genotype Real-time polymerase chain reaction Minas Gerais, Brazil |
title_short |
MELTING CURVE ANALYSIS FOR THE SCREENING OF HEPATITIS B VIRUS GENOTYPES A, D AND F IN PATIENTS FROM A GENERAL HOSPITAL IN SOUTHERN BRAZIL |
title_full |
MELTING CURVE ANALYSIS FOR THE SCREENING OF HEPATITIS B VIRUS GENOTYPES A, D AND F IN PATIENTS FROM A GENERAL HOSPITAL IN SOUTHERN BRAZIL |
title_fullStr |
MELTING CURVE ANALYSIS FOR THE SCREENING OF HEPATITIS B VIRUS GENOTYPES A, D AND F IN PATIENTS FROM A GENERAL HOSPITAL IN SOUTHERN BRAZIL |
title_full_unstemmed |
MELTING CURVE ANALYSIS FOR THE SCREENING OF HEPATITIS B VIRUS GENOTYPES A, D AND F IN PATIENTS FROM A GENERAL HOSPITAL IN SOUTHERN BRAZIL |
title_sort |
MELTING CURVE ANALYSIS FOR THE SCREENING OF HEPATITIS B VIRUS GENOTYPES A, D AND F IN PATIENTS FROM A GENERAL HOSPITAL IN SOUTHERN BRAZIL |
author |
BECKER,Carlos Eduardo |
author_facet |
BECKER,Carlos Eduardo KRETZMANN,Nelson Alexandre MATTOS,Angelo Alves de VEIGA,Ana Beatriz Gorini da |
author_role |
author |
author2 |
KRETZMANN,Nelson Alexandre MATTOS,Angelo Alves de VEIGA,Ana Beatriz Gorini da |
author2_role |
author author author |
dc.contributor.author.fl_str_mv |
BECKER,Carlos Eduardo KRETZMANN,Nelson Alexandre MATTOS,Angelo Alves de VEIGA,Ana Beatriz Gorini da |
dc.subject.por.fl_str_mv |
Hepatitis B virus Melting Curve Analysis Genotype Real-time polymerase chain reaction Minas Gerais, Brazil |
topic |
Hepatitis B virus Melting Curve Analysis Genotype Real-time polymerase chain reaction Minas Gerais, Brazil |
description |
Context Hepatitis B virus (HBV) can cause fulminant hepatitis, cirrhosis and hepatocellular carcinoma, and is one of the most common causes of acute and chronic liver failure. The genetic variants of HBV can be decisive for the evolution of these diseases as well as for the election of therapy. Objectives The aim of this study was to evaluate and standardize an in house methodology based on the analysis of the melting curve polymerase chain reaction (PCR) of real-time (qPCR) to screen for genotypes A, D and F of HBV in patients from a hospital in Rio Grande do Sul, Brazil. Methods We evaluated 104 patients presumably with HBV chronic infection. Viral DNA was extracted from plasma and viral genotypes and different mutations were determined using PCR-based protocols. Results A PCR-based methodology was standardized for the analysis of genotypes A, D and F of HBV. The technique was based in a nested PCR with the final step consisting of a multiplex real-time PCR, using the melting curve as a tool for the differentiation of fragments. A higher frequency of genotype D (44.4%), followed by genotype A (22.2%) and genotype F (3.7%) was observed. Conclusion The standardized assay, a nested PCR-multiplex qPCR using specific primers, provides a rapid and accurate method for the differentiation of HBV genotypes that are more frequent in Southern Brazil – A, D and F. This method can be applied in the clinical practice. |
publishDate |
2013 |
dc.date.none.fl_str_mv |
2013-09-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0004-28032013000300219 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0004-28032013000300219 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/S0004-28032013000200039 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Instituto Brasileiro de Estudos e Pesquisas de Gastroenterologia e Outras Especialidades - IBEPEGE. |
publisher.none.fl_str_mv |
Instituto Brasileiro de Estudos e Pesquisas de Gastroenterologia e Outras Especialidades - IBEPEGE. |
dc.source.none.fl_str_mv |
Arquivos de Gastroenterologia v.50 n.3 2013 reponame:Arquivos de gastroenterologia (Online) instname:Instituto Brasileiro de Estudos e Pesquisas de Gastroenterologia instacron:IBEPEGE |
instname_str |
Instituto Brasileiro de Estudos e Pesquisas de Gastroenterologia |
instacron_str |
IBEPEGE |
institution |
IBEPEGE |
reponame_str |
Arquivos de gastroenterologia (Online) |
collection |
Arquivos de gastroenterologia (Online) |
repository.name.fl_str_mv |
Arquivos de gastroenterologia (Online) - Instituto Brasileiro de Estudos e Pesquisas de Gastroenterologia |
repository.mail.fl_str_mv |
||secretariaarqgastr@hospitaligesp.com.br |
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1754193346917564416 |