Epidemiology and genotype analysis of emerging sapovirus-associated infections across Europe.

Sapoviruses (SaVs) belong to the Caliciviridae family and can cause gastroenteritis in humans and swine. Despite extensive testing, human sapoviruses have been found only in sporadic cases and in one mixed outbreak in children between 1994 and 2007 in the Netherlands. Here we describe a change in sapovirus epidemiology in the Netherlands resulting in sapovirus outbreaks and infections in adults. From November 2007 to January 2009, 478 outbreaks of acute gastroenteritis were reported to the National Institute for Public Health and the Environment in the Netherlands as a part of ongoing surveillance. Sapoviruses were found to be the most likely cause of 19 outbreaks (4%). During the same 2-year period, sapovirus infections were reported in Sweden, Slovenia, and Hungary. In the Netherlands, further characterization of outbreak strains showed that 12 (63%) sapovirus outbreaks were caused by genotype I.2 viruses. Most patients were adults older than 60 years (range, 1 to 100 years). Phylogenetic analysis using all presently available SaV sequences showed high homology between genotype I.2 strains detected in different geographical regions (Sweden, Slovenia, Taiwan, Japan, and Russia) since 2007. These first reported outbreaks of sapovirus infections in adults in the Netherlands were remarkable. Detection of identical genotypes in many samples might suggest that these viruses have the same origin, and since the infection is spreading fast, the prevalence of sapovirus infection may be increasing. The incidence of sapovirus infections in these countries suggests that a substantial part of Europe is affected by this virus.

Genetic diversity among food-borne and waterborne norovirus strains causing outbreaks in Sweden.

A total of 101 food-borne and waterborne outbreaks that were caused by norovirus and that resulted in more than 4,100 cases of illness were reported to the Swedish Institute for Infectious Disease Control from January 2002 to December 2006. Sequence and epidemiological data for isolates from 73 outbreaks were analyzed. In contrast to health care-related outbreaks, no clear seasonality could be observed. Sequence analysis showed a high degree of genetic variation among the noroviruses detected. Genogroup II (GII) viruses were detected in 70% of the outbreaks, and of those strains, strains of GII.4 were the most prevalent and were detected in 25% of all outbreaks. The GII.4 variants detected in global outbreaks in health care settings during 2002, 2004, and 2006 were also found in the food-borne outbreaks. GI strains totally dominated as the cause of water-related (drinking and recreational water) outbreaks and were found in 12 of 13 outbreaks. In 14 outbreaks, there were discrepancies among the polymerase and capsid genotype results. In four outbreaks, the polymerase of the recombinant GII.b virus occurred together with the GII.1 or GII.3 capsids, while the GII.7 polymerase occurred together with the GII.6 and GII.7 capsids. Mixed infections were observed in six outbreaks; four of these were due to contaminated water, and two were due to imported frozen berries. Contaminated food and water serve as important reservoirs for noroviruses. The high degree of genetic diversity found among norovirus strains causing food-borne and waterborne infections stresses the importance of the use of broad reaction detection methods when such outbreaks are investigated.

Norovirus strains belonging to the GII.4 genotype dominate as a cause of nosocomial outbreaks of viral gastroenteritis in Sweden 1997--2005. Arrival of new variants is associated with large nation-wide epidemics.

BACKGROUND: In recent years an increase of the incidence of nosocomial outbreaks caused by noroviruses has been observed throughout Sweden, with high peaks noted in the winter seasons 2002/2003 and 2004/2005, respectively. OBJECTIVES: To phylogenetically characterize norovirus strains causing nosocomial outbreaks from 1997 to 2005 and estimate the impact of norovirus-like disease on the Swedish health care system during the peak season 2002/2003 when a new variant of norovirus occurred. STUDY DESIGN: Stool samples from 115 randomly selected nosocomial outbreaks occurring during 1997--2005 throughout Sweden were studied by RT-PCR and sequencing. In addition, to investigate the impact on the health-care system, a questionnaire was distributed to infection control units (n=90) serving all Swedish hospitals, nursing homes and other health-care institutions during the largest epidemic of nosocomial outbreaks. RESULTS: Sequencing of 279 nucleotides of the norovirus RNA polymerase gene in stools containing norovirus RNA showed that strains belonging to the GII.4 genotype dominated. Each of the two large epidemics was due to a new variant within this cluster. The questionnaire revealed that 30,000-35,000 episodes of nosocomial norovirus-like infections occurred in 80 of 82 major Swedish hospitals affected in 2002/2003. CONCLUSION: New norovirus variants within the cluster GGII.4 may have a major impact on the health-care system.

A nosocomial sapovirus-associated outbreak of gastroenteritis in adults.

The human caliciviruses norovirus and sapovirus are leading causes of acute, non-bacterial gastroenteritis. In contrast to norovirus, sapovirus is known to give infections mainly in infants and young children. We describe a nosocomial outbreak of gastroenteritis associated with sapovirus involving 23 adult patients and medical staff. The mean age of the patients and medical staff was 52 y and the major signs and symptoms were nausea, diarrhoea, vomiting, abdominal cramp, headache, myalgia and fever. More patients had diarrhoea (72%) than vomiting (56%) and the mean duration of symptoms was 6 d. A secondary attack rate of 45% was seen affecting in all 10 persons with a mean age of 29 y. Sequences of the capsid gene revealed a 97% nucleotide homology to the sapovirus genogroup IV reference strain Chiba/000671T/1999. This is one of the first reported nosocomial outbreaks of sapovirus infection among adults and shows that a diagnostic test for sapovirus should be included in investigation of gastroenteritis among adults.

Evolution of human calicivirus RNA in vivo: accumulation of mutations in the protruding P2 domain of the capsid leads to structural changes and possibly a new phenotype.

In the present study we report on evolution of calicivirus RNA from a patient with chronic diarrhea (i.e., lasting >2 years) and viral shedding. Partial sequencing of open reading frame 1 (ORF1) from 12 consecutive isolates revealed shedding of a genogroup II virus with relatively few nucleotide changes during a 1-year period. The entire capsid gene (ORF2) was also sequenced from the same isolates and found to contain 1,647 nucleotides encoding a protein of 548 amino acids with similarities to the Arg320 and Mx strains. Comparative sequence analysis of ORF2 revealed 32 amino acid changes during the year. It was notable that the vast majority of the cumulative amino acid changes (8 of 11) appeared within residues 279 to 405 located within the hypervariable domain (P2) of the capsid protein and hence were subject to immune pressure. An interesting and novel observation was that the accumulated amino acid changes in the P2 domain resulted in predicted structural changes, including disappearance of a helix structure, and thus a possible emergence of a new phenotype. FUT2 gene polymorphism characterization revealed that the patient is heterozygous at nucleotide 428 and thus Secretor(+), a finding in accordance with the hypothesis of FUT2 gene polymorphism and calicivirus susceptibility. To our knowledge, this is the first report of RNA evolution of calicivirus in a single individual, and our data suggest an immunity-driven mechanism for viral evolution. We also report on chronic virus excretion, immunoglobulin treatment, and modification of clinical symptoms; our observations from these studies, together with the FUT2 gene characterization, may lead to a better understanding of calicivirus pathogenesis.