György Szücs

Increase in viral gastroenteritis outbreaks in Europe and epidemic spread of new norovirus variant

BACKGROUND Highly publicised outbreaks of norovirus gastroenteritis in hospitals in the UK and Ireland and cruise ships in the USA sparked speculation about whether this reported activity was unusual. METHODS We analysed data collected through a collaborative research and surveillance network of viral gastroenteritis in ten European countries (England and Wales were analysed as one region). We compiled data on total number of outbreaks by month, and compared genetic sequences from the isolated viruses. Data were compared with historic data from a systematic retrospective review of surveillance systems and with a central database of viral sequences. FINDINGS Three regions (England and Wales, Germany, and the Netherlands) had sustained epidemiological and viral characterisation data from 1995 to 2002. In all three, we noted a striking increase in norovirus outbreaks in 2002 that coincided with the detection and emergence of a new predominant norovirus variant of genogroup II4, which had a consistent mutation in the polymerase gene. Eight of nine regions had an annual peak in 2002 and the new genogroup II4 variant was detected in nine countries. Also, the detection of the new variant preceded an atypical spring and summer peak of outbreaks in three countries. INTERPRETATION Our data from ten European countries show a striking increase and unusual seasonal pattern of norovirus gastroenteritis in 2002 that occurred concurrently with the emergence of a novel genetic variant. In addition to showing the added value of an international network for viral gastroenteritis outbreaks, these observations raise questions about the biological properties of the variant and the mechanisms for its rapid dissemination.

Analysis of Integrated Virological and Epidemiological Reports of Norovirus Outbreaks Collected within the Foodborne Viruses in Europe Network from 1 July 2001 to 30 June 2006

ABSTRACT The Foodborne Viruses in Europe network has developed integrated epidemiological and virological outbreak reporting with aggregation and sharing of data through a joint database. We analyzed data from reported outbreaks of norovirus (NoV)-caused gastroenteritis from 13 European countries (July 2001 to July 2006) for trends in time and indications of different epidemiology of genotypes and variants. Of the 13 countries participating in this surveillance network, 11 were capable of collecting integrated epidemiological and virological surveillance data and 10 countries reported outbreaks throughout the entire period. Large differences in the numbers and rates of reported outbreaks per country were observed, reflecting the differences in the focus and coverage of national surveillance systems. GII.4 strains predominated throughout the 5-year surveillance period, but the proportion of outbreaks associated with GII.4 rose remarkably during years in which NoV activity was particularly high. Spring and summer peaks indicated the emergence of genetically distinct variants within GII.4 across Europe and were followed by increased NoV activity during the 2002-2003 and 2004-2005 winter seasons. GII.4 viruses predominated in health care settings and in person-to-person transmission. The consecutive emergence of new GII.4 variants is highly indicative of immune-driven selection. Their predominance in health care settings suggests properties that facilitate transmission in settings with a high concentration of people such as higher virus loads in excreta or a higher incidence of vomiting. Understanding the mechanisms driving the changes in epidemiology and clinical impact of these rapidly evolving RNA viruses is essential to design effective intervention and prevention measures.

Sequence heterogeneity among human picobirnaviruses detected in a gastroenteritis outbreak

Summary. Human picobirnaviruses characterised in this study were serendipitously detected in a non-bacterial gastroenteritis outbreak when specimens were examined for the presence of human rotaviruses using polyacrylamide gel electrophoresis. Of ten stool samples sent for virological examination, two, three, and one specimens were positive for human caliciviruses, picobirnaviruses, and both viruses, respectively. Partial sequences of the RNA-dependent RNA polymerase gene were determined for three picobirnavirus-positive samples. The sequence identity among these three strains was 60% to 65% for the nucleic acid and 64% to 70% for the deduced amino acid sequences. Phylogenetic analysis revealed that each of the three strains clustered with strains identified in geographically separate areas. In contrast, human calicivirus strains co-incidentally identified, showed complete nucleotide sequence identity. These findings demonstrate a lack of common exposure to or point of source for picobirnavirus infection, suggesting that the outbreak was caused by human caliciviruses. Further studies are needed to determine the etiologic role and to establish the taxonomic basis of picobirnaviruses.

Eight-Year Survey of Human Rotavirus Strains Demonstrates Circulation of Unusual G and P Types in Hungary

ABSTRACT Between 1992 and 2000, a total of 4,173 rotavirus-positive samples were collected from two areas of Hungary. Of these, 2,020 specimens (48.4%) were analyzed for G serotype, using monoclonal antibody-based immunoassay and reverse transcription-PCR. By the two methods, 1,789 samples were specified as G1 (62%), G2 (12.2%), G3 (1.4%), G4 (6.4%), G6 (1.0%), G9 (2.9%), or mixed infection (2.6%), and the remaining 231 (11.4%) could not be G typed. The linkage between G and P type, subgroup specificity, and RNA profile was investigated with a sample subset. Among these specimens, we identified both the four globally common strains (P[8],G1 subgroup II (sgII); P[4],G2 sgI; P[8],G3 sgII; and P[8],G4 sgII) and six uncommon strains (P[6],G4 sgII; P[9],G3 sgI; P[9],G6 sgI; P[14],G6 sgI; P[8],G9 sgII; and P[8],G9 sgI). All strains with P[8], P[6], P[9], and P[14] specificities had a long electropherotype, whereas most of those carrying a P[4] specificity were associated with a short electropherotype. Although once considered to be rare, P[9],G6 and P[8],G9 rotavirus strains represent potentially important new serotypes in Hungary.

Genogroup I picobirnaviruses in pigs: Evidence for genetic diversity and relatedness to human strains

Picobirnaviruses (PBVs) are small, non-enveloped viruses with a bisegmented double-stranded RNA genome. Their pathogenic potential, ecology, and evolutionary features are largely unexplored. Here, we describe the molecular analysis of porcine PBVs identified in the intestinal content of dead pigs. Six of 13 positive samples were cloned and then subjected to single-strand conformation polymorphism analysis and nucleotide sequencing. All clones belonged to genogroup I PBVs and almost all clones clustered on separate branches from human strains. A single strain shared a notably close genetic relationship with a Hungarian human PBV strain (89.9 nt and 96.4% aa identity). Genetic diversity was also observed among strains identified in mixed infections. Single point mutations and deleterious mutations within highly related strains suggested that PBVs exist as quasispecies in the swine alimentary tract. Clones with complete sequence identities originating from different animals suggested effective animal-to-animal transmission of the virus. Our findings indicate that infection with genogroup I PBVs is common in pigs.

Emergence of serotype G12 rotaviruses, Hungary.

We describe the emergence of serotype G12 rotaviruses (67 [6.9%] of 971 specimens tested) among children hospitalized with rotavirus gastroenteritis in Hungary during 2005. These findings are consistent with recent reports of the possible global spread and increasing epidemiologic importance of these strains, which may have implications for current rotavirus vaccination strategies.

Sequencing and Phylogenetic Analysis of Human Genotype P[6] Rotavirus Strains Detected in Hungary Provides Evidence for Genetic Heterogeneity within the P[6] VP4 Gene

ABSTRACT Although rotavirus genotype P[6] is one of the three most common VP4 specificities associated with human infection, the relatively few sequence data available in public databases suggest that the genetic variability within P[6] might be presently unexplored. Thus far, two human P[6] lineages (M37-like and AU19-like) and a single porcine P[6] lineage (Gottfried-like) have been identified by phylogenetic analysis. Serologic studies demonstrated that these three lineages are antigenically distinct from each other, a finding based on which they were classified into three subtypes, P2A[6] (M37-like), P2B[6] (Gottfried-like), and P2C[6] (AU19-like). To study heterogeneity within this genotype, we selected for molecular characterization a total of six P[6] strains detected during an ongoing surveillance in Hungary. The variable region of the VP4 gene was subjected to sequencing and phylogenetic analysis. Our data indicated that these six strains fell into two phylogenetic lineages distinguishable from the human lineages M37-like and AU19-like and from the porcine lineage Gottfried-like. Further studies are needed to understand whether these two novel lineages are genuine human strains or might have originated from animal strains and to evaluate the antigenic relationship of the novel Hungarian P[6] strains to the three established subtypes.

Enteric caliciviruses in domestic pigs in Hungary

Summary.Caliciviruses closely related to human norovirus and sapovirus were recently detected in domestic pigs, causing discussions about the animal reservoir and the potential for zoonotic transmission to humans. To detect porcine caliciviruses, 17 fecal samples collected on two swine farms in southwestern Hungary were tested by reverse transcription-polymerase chain reaction. Three (17.6%) samples were positive for caliciviruses. This study confirms the presence of caliciviruses, both porcine sapovirus (genus Sapovirus) and porcine norovirus (genus Norovirus), in domestic pigs in Hungary and provides additional information on the viral genetic diversity and relationship to viruses referred to as human caliciviruses.

Reovirus identified as cause of disease in young geese.

The pathology, epizootiology and aetiology of a specific disease of young geese, which has been seen in Hungary for more than three decades, were investigated. The disease was characterised by splenitis and hepatitis with miliary necrotic foci during the acute phase, and epicarditis, arthritis and tenosynovitis during the subacute/chronic phase. Clinical signs usually appeared at 2 to 3 weeks of age and persisted for 3 to 6 weeks. From different organs of the affected birds, a reovirus was isolated in embryonated eggs and tissue cultures ofMuscovy duck or goose origin, as well as in Vero cells. In experimental infections, the dominant features of thedisease were reproduced in day-old and young goslings. The biological and partial molecular characterisation ofone of the isolated strains (D15/99) showed that it was related to the reovirus described as the cause of a similardisease of Muscovy ducks. An RT-PCR method suitable for the detection of reoviruses was also elaborated andtested. This is the first report on the involvement of reovirus in arthritis of geese.

Detection of human rotavirus serotype G6 in Hungary

During an ongoing survey of human rotavirus serotypes, we demonstrated for the first time the circulation of serotype G6 in two regions of Hungary. Of five rotavirus seasons surveyed to date (1994-9), serotype G6 was found in all seasons except 1994-5 at an overall prevalence of 1.4% (17 of 1252) and ranging from 0.6 to 4.5%. Children infected with G6 strains were older (mean age, 3.3 years) than children infected with the four (G1-G4) globally common serotypes (mean age, 2.1 years; unpaired Students t test, P<0.001). Our data indicate that rotavirus serotype G6 may be an epidemiologically important G serotype in Hungary.