Molecular characterization of hepatitis A virus isolates from environmental and clinical samples in Greece.
BACKGROUND
Hepatitis A virus (HAV) strains detected in environmental and clinical samples were analysed to characterize the genotypes of HAV circulating in Greece. Fifty (50) sewage samples were collected from Patras (South-Western Greece) and Alexandroupolis (North-Eastern Greece) from 2007 until 2009, accordingly. The clinical samples derived from an HAV outbreak involved populations from three neighbouring prefectures of North-Eastern Greece (Xanthi, Rodopi, and Evros). HAV particles were detected by nested RT-PCR, using a previously validated set of primers to amplify a 290-bp fragment encompassing the 5'-NTR. Positive HAV samples were confirmed by sequencing of the PCR product. To determine the relatedness between the different isolated sequences, a phylogenetic tree was constructed.


RESULTS
Results showed a 100% prevalence of genotype I, and particularly subgenotype IA. The analyzed HAV strains were closely related between them with the percentage of nucleotide identity ranging between 96% and 100%.


CONCLUSIONS
The study revealed the major prevalence of circulating strains of IA genotype in Greece and underlined the usefulness of molecular methods for the detection and typing of viruses in both environmental and clinical samples. The present study is, to our knowledge, the first in Greece to depict the simultaneous molecular characterization of HAV strains isolated from both clinical and environmental samples.
Kokkinos
Petros
P
Environmental Microbiology Unit, Department of Public Health, School of Medicine, University of Patras, Greece.
Ziros
Panos
P
Filippidou
Sevasti
S
Mpampounakis
Ioannis
I
Vantarakis
Apostolos
A
D020121:5' Untranslated Regions; D016000:Cluster Analysis; D004196:Disease Outbreaks; D004783:Environmental Microbiology; D005838:Genotype; D006115:Greece / Q000453:epidemiology; D006506:Hepatitis A / Q000453:epidemiology* / Q000821:virology*; D030041:Hepatitis A virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D010802:Phylogeny; D012367:RNA, Viral / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid
D020121:5' Untranslated Regions; D012367:RNA, Viral