Hepatitis C: genotyping

Cavalheiro, Norma de Paula

doi:10.1590/s1413-86702007000700009

Cited by 4 publications

(3 citation statements)

References 18 publications

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“…HCV genotyping methods include PCR amplification of the target gene and sequencing, PCR amplification and hybridization with genotype-specific probes, and real-time RT-PCR. 23 Currently, there are no FDA-approved assays to determine HCV genotypes. The reference method for HCV genotyping is PCR amplification and direct sequencing of HCV genomic NS5B or 5'UTR regions, alignments, and phylogenetic analysis.…”

Section: Genotyping Techniquesmentioning

confidence: 99%

“…However, the advantage of direct sequencing is that it reveals genomic variability and the presence of quasi-species during the natural progression of disease and its response to antiviral therapy. 23 In clinical practice, HCV genotyping is performed using commercially available kits that employ PCR amplification and hybridization with genotype-specific probes. Currently used methods include reverse-hybridization line probe INNO-LiPA HCV II assay (Innogenetics, Ghent, Belgium), simplified direct sequencing Trugene 5'NC HCV Genotyping assays (Siemens AG), and the Abbott Real-Time HCV Genotype II Assay (Abbott Laboratories) ( Table 3).…”

Section: Genotyping Techniquesmentioning

confidence: 99%

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Laboratory Methods for Diagnosis and Management of Hepatitis C Virus Infection

Amjad¹,

Moudgal²,

Muhammad³

2013

Lab Med

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Hepatitis C virus (HCV) infection is associated with chronic hepatitis, cirrhosis, and hepatocellular carcinoma. This review summarizes the pathogenesis and significance of serological and molecular-based assays in the diagnosis and management of HCV infection.After reading this article, readers should be able to describe laboratory tests used in the diagnosis and management of HCV infection. They should also be able to describe the general principles for selecting the most appropriate laboratory test for diagnosis, therapeutic decision making, and assessment of virologic response to therapy.

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Section: Genotyping Techniquesmentioning

confidence: 99%

Section: Genotyping Techniquesmentioning

confidence: 99%

Laboratory Methods for Diagnosis and Management of Hepatitis C Virus Infection

Amjad¹,

Moudgal²,

Muhammad³

2013

Lab Med

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“…A genotipagem por hibridização reversa, distribuída sob o nome comercial de Line Probe Assay (INNO-LiPA ® ) (Siemens Medical Solutions Diagnostics), é muito utilizada em vários países. Este teste utiliza sondas específicas para diferentes partes do genoma viral imobilizadas em uma membrana de nitrocelulose 2,3 . Existem atualmente duas versões do LiPA (1.0 e 2.0).…”

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Hibridização reversa e sequenciamento na genotipagem do vírus da hepatite C

Levada

Moraes

Corvino

et al. 2010

Rev. Soc. Bras. Med. Trop.

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RESUMOIntrodução: Os métodos de genotipagem do vírus da hepatite C têm sido muito discutidos. O objetivo deste trabalho foi comparar as metodologias de hibridização reversa e sequenciamento direto para a genotipagem do vírus da hepatite C. Métodos: Noventa e uma amostras de plasma de pacientes assistidos na Faculdade de Medicina de Botucatu da Universidade Estadual Paulista foram utilizadas. A genotipagem por hibridização reversa foi realizada utilizando o kit comercial INNO-LiPA  v.1.0. O sequenciamento direto foi efetuado em sequenciador automático utilizando protocolos in house. Resultados: A genotipagem por sequenciamento direto mostrou-se eficiente na resolução dos resultados inconclusivos pelo kit comercial. O kit mostrou resultados errôneos em relação à subtipagem viral. Além disso, a genotipagem por sequenciamento direto revelou um erro do kit com relação à determinação genotípica questionando a eficiência do método também para a identificação do genótipo viral. Conclusões: A genotipagem realizada por meio de sequenciamento direto permite uma maior acurácia na classificação viral quando comparada à hibridização reversa Palavras-chaves: Vírus da hepatite C. Genotipagem. Hibridização reversa. Sequenciamento direto. ABSTRACT Introduction:The methods for genotyping the hepatitis C virus have been much discussed. The aim of this study was to compare the methodologies of reverse hybridization and direct sequencing for genotyping the hepatitis C virus. Methods: Ninety-one plasma samples from patients attended at the Botucatu Medical School, São Paulo State University, were used. Genotyping by reverse hybridization was performed using the INNO-LiPA  v.1.0 commercial kit. Direct sequencing was performed in an automated sequencer using in-house protocols. Results: Genotyping by direct sequencing was shown to be efficient for resolving cases that had remained inconclusive after using the commercial kit. The kit showed erroneous results in relation to virus subtyping. Moreover, direct sequencing revealed an error of the kit regarding the genotypic determination, thereby raising doubts about the efficiency of reverse hybridization for identifying the virus genotype. Conclusions: Genotyping by direct sequencing allowed greater accuracy of virus classification than did reverse hybridization.

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Molecular Epidemiological Study of Hepatitis Viruses in Ismailia, Egypt

Youssef¹,

Yano

Utsumi³

et al. 2009

Intervirology

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The hepatitis virus is hyperendemic in Egypt, western Asia and Africa. In Egypt, most studies have been carried out in the regions of the upper and lower Nile Delta, and so little is known about other parts of the country. Our project aimed to clarify the carrier rate of various hepatitis viruses in the northeastern province of Ismailia. A total of 214 patients with elevated liver enzymes were enrolled in this study. Sera were collected in Ismailia hospital. We conducted a serological and molecular-based survey of hepatitis viruses, including their genetic variability and genotype-related differences. There were 10 (4.7%) cases positive for hepatitis B surface antigen (HBsAg), and 156 (72.9%) positive for anti-hepatitis C virus antibody (HCV-Ab). Phylogenetic analysis showed that genotype C of HBV and genotype 4a of HCV were prevalent. Hepatitis D virus RNA was not detected in HBsAg carriers. Although anti-hepatitis E virus IgM antibody was positive in 5 cases (2.3%), no case was positive for its RNA. Among 54 cases negative for HBsAg and HCV-Ab, HBV-DNA was detected in 35 cases (65%). Our results revealed that HBV and HCV, including occult HBV infection, are widespread and related to liver diseases in Ismailia province, Egypt.

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Hepatitis C: genotyping

Cited by 4 publications

References 18 publications

Laboratory Methods for Diagnosis and Management of Hepatitis C Virus Infection

Laboratory Methods for Diagnosis and Management of Hepatitis C Virus Infection

Hibridização reversa e sequenciamento na genotipagem do vírus da hepatite C

Molecular Epidemiological Study of Hepatitis Viruses in Ismailia, Egypt

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