Molecular epidemiology of hepatitis C virus in Romania
Abstract number: P1905
Oprisan G., Szmal C., Dinu S., Oprisoreanu A., Thiers V.
Objectives: To evaluate a home-made genotyping method for investigation of Hepatitis C virus (HCV) variability and tracing infections in Romania.
Methods: sera from Romanian patients were analysed by RT-PCR and sequencing in the core region. PCR amplicons with degenerated primers were sequenced using an automated sequencer ABI Prism 3100 Avant (Applied Biosystems). Multiple sequence alignments and translation into amino acid sequences were performed with BioEdit programme. NCBI BLAST database was screened for similar sequences. Phylogenetic and molecular evolutionary analyses were conducted using MEGA version 4.0 with the neighbour-joining method. The distance matrix was calculated by the Kimura two-parameter method using DNADIST.
Results: Two hundred sera from Romanian patients were analysed in the core region using a home-made semi-nested PCR and sequencing. Genotype distribution was: 1b 95%, 1a 3%, 4a 1.5% and 3a 0.5%. Within subtype 1b, the Romanian strains were intermixed with other 1b strains circulating in European countries. Genetic distances between six 1a strains isolated from injecting drug users revealed a single source of infection. The 4a strains presented 97% homology with the strain ED43 from Egypt. All the core sequences obtained presented an alternative core reading frame. A nosocomial outbreak of HCV infection in a urology clinic from Cluj was investigated using the core sequence analysis. The analysis of 300 nucleotides in the core region allowed identification of a transmission event among the medical staff, involving one source and two infected persons. The core sequences from three patients were monophyletic in the core phylogenetic tree and presented genetic distances of 0.0040.007 (Kimura two-parameter method). The other unrelated 1b strains presented genetic distances of 0.040 from the outbreak strains.
Conclusion: The new RT-PCR in the core region has a good resolving power for HCV subtyping and evaluating transmission routes. The predominant HCV subtype in Romania is 1b, while subtypes 1a and 3a might have been recently introduced in Romania as a result of drug abuse in the younger population.
|Session name:||18th European Congress of Clinical Microbiology and Infectious Diseases|
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