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A Protocol for Analyzing Hepatitis C Virus Replication
Published on: June 26, 2014
Distribution and molecular characterization of hepatitis C virus (HCV) genotypes in patients with chronic infection
Mónica V Alvarado-Mora1, Izolda M Moura, Livia S Botelho-Lima
1Laboratory of Tropical Gastroenterology and Hepatology, Department of Gastroenterology, School of Medicine, University of São Paulo, São Paulo, Brazil. monica.viviana@usp.br
Insights
Hepatitis C virus (HCV) genotype 1b is the most common in Brazil's Northeast, affecting 42-66.7% of patients. This study characterized circulating HCV genotypes in Pernambuco State, Brazil.
Area of Science:
- Virology
- Epidemiology
- Public Health
Background:
- Hepatitis C virus (HCV) poses a global health challenge, infecting 3% of the world's population annually.
- Brazil faces a significant HCV burden, with the Northeast region exhibiting the highest prevalence.
Purpose of the Study:
- To determine the prevalent Hepatitis C virus genotypes in Pernambuco State, Brazil.
- To contribute to understanding HCV epidemiology in Brazil's diverse population.
Main Methods:
- Retrospective analysis of 85 anti-HCV positive patients from 2004-2011.
- HCV RNA NS5B region amplification using nested PCR.
- Phylogenetic analysis via Bayesian Markov chain Monte Carlo simulation.
Main Results:
- Successfully sequenced 63 (74.1%) of the samples.
- HCV genotype 1b was the most prevalent (42-66.7%), followed by 3a (16-25.4%), 1a (4-6.3%), and 2b (1-1.6%).
- Among patients with co-infection, genotype 1b and 3a were observed in 63.1% and 36.9% respectively.
Conclusions:
- Hepatitis C virus genotype 1b is the predominant strain in Pernambuco State, Brazil.
- Phylogenetic analysis indicates the circulation of diverse HCV strains within the studied population.
- Findings highlight regional variations in HCV genotypes across Brazil.
Abstract:
Hepatitis C virus (HCV) is a public health problem throughout the world and 3% of the world population is infected with this virus. It is estimated that 3-4 millions individuals are being infected every year. It has been estimated that around 1.5% of Brazilian population is anti-HCV positive and the Northeast region showed the highest prevalence in Brazil. The aim of this study was to characterize HCV genotypes circulating in Pernambuco State (PE), Brazil, located in the Northeast region of the country. This study included 85 anti-HCV positive patients followed up between 2004 and 2011. For genotyping, a 380bp fragment of HCV RNA in the NS5B region was amplified by nested PCR. Phylogenetic analysis was conducted using Bayesian Markov chain Monte Carlo simulation (MCMC) using BEAST v.1.5.3. From 85 samples, 63 (74.1%) positive to NS5B fragment were successfully sequenced. Subtype 1b was the most prevalent in this population (42-66.7%), followed by 3a (16-25.4%), 1a (4-6.3%) and 2b (1-1.6%). Twelve (63.1%) and seven (36.9%) patients with HCV and schistosomiasis were infected with subtypes 1b and 3a, respectively. Brazil is a large country with many different population backgrounds; a large variation in the frequencies of HCV genotypes is predictable throughout its territory. This study reports HCV genotypes from Pernambuco State where subtype 1b was found to be the most prevalent. Phylogenetic analysis suggests the presence of the different HCV strains circulating within this population.
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