UBC Faculty Research and Publications

Molecular characterization of rotavirus isolates from select Canadian pediatric hospitals McDermid, Andrew; Le Saux, Nicole; Grudeski, Elsie; Bettinger, Julie A.; Manguiat, Kathy; Halperin, Scott A.; MacDonald, Lily; Déry, Pierre; Embree, Joanne; Vaudry, Wendy; Booth, Timothy F. Nov 15, 2012

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RESEARCH ARTICLEMolecular characterizationlieVaPrenteritis in children under five, causing an estimated453,000 deaths worldwide annually [1,2]. In Canada, asas lost work days that have a substantial economiccost [3-6].McDermid et al. BMC Infectious Diseases 2012, 12:306http://www.biomedcentral.com/1471-2334/12/306genotypes and 12 P genotypes have been recoveredfrom humans [10-14]. G and P genotypes are also 2CanadaFull list of author information is available at the end of the articlein other developed countries, mortality is rare, althoughrotavirus infections are costly from both societal andhealth care system perspectives. Recently publishedresearch on children with gastroenteritis admittedto Canadian paediatric hospitals found that rotavirusinfections require 1300–1800 tertiary care hospital daysThe outer capsid of rotavirus is composed of twomajor antigenic proteins, VP4 and VP7. These proteinsare the main determinants of viral serotype, and thegenes that code for these proteins represent the P andG genotype of rotaviruses respectively. The VP4 andVP7 proteins are also the main targets for protectiveneutralizing antibodies, and are thus key antigens invaccine development [7-9]. To date, 27 G genotypes and35 P genotypes have been reported, although only 11 G* Correspondence: tim.booth@phac-aspc.gc.ca1National Microbiology Laboratory, Winnipeg, Canada2Department of Medical Microbiology, University of Manitoba, Winnipeg,Background: We report the first multi-site rotavirus genotype analysis in Canada. Prior to this study, there was adearth of rotavirus G and P genotyping data in Canada. Publically funded universal rotavirus vaccination in Canadastarted in 2011 and has been introduced by four provinces to date. Uptake of rotavirus vaccines in Canada prior to2012 has been very limited. The aim of this study was to describe the genotypes of rotavirus strains circulating inCanada prior to widespread implementation of rotavirus vaccine by genotyping samples collected from selectedpaediatric hospitals. Secondly we identified rotavirus strains that differed genetically from those included in thevaccines and which could affect vaccine effectiveness.Methods: Stool specimens were collected by opportunity sampling of children with gastroenteritis who presentedto emergency departments. Samples were genotyped for G (VP7) genotypes and P (VP4) genotypes byhemi-nested multiplex PCR methods. Phylogenetic analysis was carried out on Canadian G9 strains to investigatetheir relationship to G9 strains that have circulated in other regions of the world.Results: 348 samples were collected, of which 259 samples were rotavirus positive and genotyped. There were 34rotavirus antigen immunoassay negative samples genotyped using PCR-based methods. Over the four rotavirusseasons, 174 samples were G1P[8], 45 were G3P[8], 22 were G2P[4], 13 were G9P[8], 3 were G4P[8] and 2 were G9P[4]. Sequence analysis showed that all Canadian G9 isolates are within lineage III.Conclusions: Although a limited number of samples were obtained from a median of 4 centres during the 4 yearsof the study, it appears that currently approved rotavirus vaccines are well matched to the rotavirus genotypesidentified at these hospitals. Further surveillance to monitor the emergence of rotavirus genotypes in Canada iswarranted.BackgroundGroup A rotaviruses are a major cause of acute gastro-annually [3]. Furthermore, rotavirus gastroenteritisresults in outpatient visits and nonmedical costs, suchfrom select Canadian pedAndrew McDermid1,2, Nicole Le Saux3, Elsie Grudeski1, JuLily MacDonald1, Pierre Déry6, Joanne Embree2,7, Wendyfor Members of the Canadian Immunization MonitoringAbstract© 2012 McDermid et al.; licensee BioMed CenCreative Commons Attribution License (http:/distribution, and reproduction in any mediumOpen Accessof rotavirus isolatesiatric hospitalsA Bettinger4, Kathy Manguiat1, Scott A Halperin5,udry8 and Timothy F Booth1,2*ogram, Active (IMPACT)tral Ltd. This is an Open Access article distributed under the terms of the/creativecommons.org/licenses/by/2.0), which permits unrestricted use,, provided the original work is properly cited.McDermid et al. BMC Infectious Diseases 2012, 12:306 Page 2 of 9http://www.biomedcentral.com/1471-2334/12/306components of the 11-component genotyping systemproposed to classify rotaviruses [14]. Five combinationsof these genotypes, G1P[8], G2P[4], G3P[8], G4P[8] andG9P[8] are responsible for the majority of rotavirusinfections worldwide. These are also the most commongenotypes in North America, where they represent morethan 85% of rotaviruses detected in human gastrointes-tinal disease [12,15-18]. Studies that have analysed onlythe G genotypes, also found that G1, G2, G3, G4 and G9predominated in North America [19-21].Two live oral rotavirus vaccines were licensed inCanada. RotaTeqW, licensed in Canada in August 2006,is a pentavalent bovine reassortant vaccine based on thebovine rotavirus WC3 strain as a backbone, and each ofthe 5 vaccine strains contains one serotype of the humanouter capsid proteins (G1, G2, G3, G4, or P[8] sero-types). RotarixW, licensed in Canada in July 2007, is amonovalent live-attenuated G1P[8]vaccine derived fromhuman rotavirus 89–12, containing the two most com-mon outer capsid serotypes. Extensive phase III trials forthese vaccines showed high efficacy in protecting chil-dren against rotavirus disease of any severity, for strainswith the same serotypes as contained in the respectivevaccine, and there was a significant degree of cross-reactivity against many genotypes not contained in thevaccines [22]. Vaccines have been shown to be very ef-fective in the United States, although changes in anti-genic properties of circulating strains, due to antigenicdrift or recombination, may challenge the effectivenessof the vaccines in the future [23-28]. In 2008, the Na-tional Advisory Committee for Immunization (NACI)and the Canadian Paediatric Society recommended theuse of rotavirus vaccine in Canada [29,30]. Despite this,the uptake of rotavirus vaccines in Canada has been verylimited. Prior to this study, there was very little data onthe incidence of rotavirus G and P genotypes in Canada.During the period of this study, vaccination use inCanada was very low, well below 5% of the eligiblepopulation were getting rotavirus vaccines (personalobservations). Starting in 2012, only four of 13 provincesand territories offered rotavirus vaccine as part of theirpublicly funded immunization programs [31].To survey the baseline prevalence of rotavirus geno-types prior to the introduction of rotavirus vaccine wecollected specimens from inpatients at select paediatrichospitals. We utilised reverse transcriptase polymerasechain reaction (RT-PCR) followed by hemi-nested multi-plex polymerase chain reaction methods for P and Ggenotyping [13,28,32-34]. Selected rotavirus strains werefurther characterised by phylogenetic analysis: in par-ticular we focussed on the Canadian G9 strains, sincethis genotype has emerged and spread worldwide in re-cent years, and is not a specific component of either ofthe current vaccines.MethodsA total of 348 stool samples were collected from childrenthat presented with diarrhea with or without vomiting toeight Canadian pediatric hospital emergency departmentsthat were part of the Canadian Immunization MonitoringProgram, Active (IMPACT) network [35] from 2007 to2010. Five pediatric hospitals were included in the studyduring the first year (2007: Edmonton, Winnipeg, Ottawa,Quebec City, Halifax), three hospitals for the years 2008and 2009 (Ottawa, Quebec City, Halifax) and six in2010 (Vancouver, Saskatoon, Winnipeg, Toronto, Ottawa,Halifax). The total number of specimens collected at thesites were as follows: Ottawa, 128; Halifax, 90; QuebecCity, 63; Edmonton, 24; Vancouver, 20; Saskatoon, 11;Winnipeg, 9; Toronto, 3.The study was approved by the research ethics boardsof each hospital, in accordance with the Helsinki Dec-laration on ethical principles for medical research in-volving human subjects. Specimens were collected fromchildren under 5 years of age that presented with diar-rhea with or without vomiting at emergency depart-ments during 2007, 2008 and 2009 [6]. Research nursesat the emergency department of each centre approachedparents or guardians of children presenting with acutediarrhea and asked them to consent to rotavirus testingon the child’s stool sample. In 2010 the samples werefrom children under 16 years of age who were hospita-lized for laboratory confirmed rotavirus infection at sixhospitals (Vancouver, Saskatoon, Winnipeg, Toronto,Ottawa, Halifax).At each hospital stool samples underwent testing forrotavirus by enzyme immunoassay (Premier rotacloneEIA kit, Meridian Bioscience), chromatographic im-munoassay (bioMerieux Vikia Rota-Adeno, or Coris Bio-concept Combi strip) or by electron microscopy (EM).All specimens were then sent to the National Microbiol-ogy Laboratory for further testing and genotyping.Genomic RNA was extracted from the stool samplesusing a Nuclisens Easymag magnetic silica extractionmethod (Biomerieux, France). Hemi-nested multiplex PCRassays were used to P and G genotype the extracted RNA[32,36-40]. PCR products were initially designated togenotype based on size comparison after direct vi-sualization after electropheresis on agarose gels. Duringthe course of genotyping work we replaced a previouslydescribed primer specific for P[8] genotypes, 1T-1 [36],with the degenerate primer 1T-1DCDN since it was foundto be complementary to a wider range of P[8] strains. The1T-1DCDN primer was designed with the sequence 5’TCT ACT GGR TYR ACR TG 3’ using the primer 1T-1D[38] as a model, and an alignment of the VP4 variable re-gion using the Clustal W algorithm in MEGA 5.0 [41].The alignment included sequences from 66 previouslyuntypable Canadian P[8] samples (determined by geneticdistance after sequencing), one Canadian P[4] sample, and15 P[8] reference strains. The 1T-1DCDN primer binds atnt 340–356, a similar position, but one nucleotide shorterthan that of 1T-1 (nt 339–356).The new 1T-1DCDN primer was compared with 1T-1primer for genotyping using a panel of 65 Canadianrotavirus samples that included 47 samples that weresequence-positive for P[8], 4 specimens that were P[4]genotype and 14 that were rotavirus negative. The panelwas representative in that it was composed of specimensthat came from 5 different sites in 2007, 3 sites in 2008and 2 sites in 2009.All strains that were positively genotyped by hemi-nested multiplex PCR were confirmed by sequencing ofthe VP4 and VP7 regions, using the Con3/Con2 andBeg9/End9 primer sets, respectively [36,37], or a suitabledesignation was based on similarity to previouslydefined lineage reference strains [42,43].ResultsStool samples were collected from children with gastro-enteritis at eight different Canadian hospital sites be-tween 2007 and 2010 (Figure 1), although only two ofthe sites were sampled in all four years, and three siteswere sampled in only one year. Of 348 specimens, 271were genotyped. Amongst these, 12 samples exhibitedmultiple genotypes, suggestive of mixed infections.Therefore, 259 specimens (74%) were assigned to a sin-gle G and P genotype (Table 1) and 77 were negative.Of the 348 samples that were screened for rotavirus inhospital, 246 (71%) were positive by antigen immuno-assay, and 102 (29%) screened negative. Of the speci-oveMcDermid et al. BMC Infectious Diseases 2012, 12:306 Page 3 of 9http://www.biomedcentral.com/1471-2334/12/306alternative. PCR products were purified using MontagePCR Centrifugal Filter Devices (Millipore, USA) andthen cloned for sequencing in Top10 chemically compe-tent E. coli cells (Invitrogen, USA) using Invitrogen5’TopoTA cloning kits or sequenced directly. Sequen-cing of plasmids was carried out by the Genomics Coresection of the National Microbiology Laboratory usingT3 and T7 plasmid-specific primers.Phylogenetic analysis was used for confirmation ofgenotypes and for further analysis of the ORFs ofCanadian G9 strains. The Canadian G9 ORF sequencesof the strains RVA/Human-wt/CAN/RT034-07/2007/G9P[8] through to RVA/Human-wt/CAN/RT088-09/2009/G9P[8] (GenBank Accession numbers JF964998-JF965010) were aligned, along with reference strains.Phylogenetic trees were constructed using the ClustalW algorithm in MEGA 5.0 software package [41], usingthe Maximum Likelihood method for phylogeneticanalysis, with 1000 bootstrap replicates. LineageFigure 1 Map showing the 8 study sites in Canada (black dots). Abgenotypes detected between the years 2007–2010. The limited Toronto datop right of the map indicates the cumulative prevalence of all genotypesmens that had screened positive in hospital 237 (96%)were positively P and G for genotyped by PCR. Of the102 specimens that had screened negative by antigen im-munoassay, 34 (33%) were also successfully P and Ggenotyped for rotavirus. Twelve (4.4%) of the rotavirus-positive samples were indeterminate since they were ap-parently mixed infections with samples containing twoor more genotypes of rotavirus (Table 1). The mixedsamples included three instances of G1P[4] + P[8] , sixinstances of G1P[8] + G2P[4] and one instance each ofG1 + G3P[8], G3P[4] + P[8] and G1 + G9P[8]. Of the 57stool samples from 2010 that were positive locally forrotavirus, 50 were successfully genotyped and confirmedby sequencing.Overall, the most common genotype encountered inthis study was G1P[8], being present in 67% of the speci-mens, and it was also the most prevalent genotype at allsites (Figure 1) and during all 4 years (Figure 2). Theother common genotypes were G3P[8] (18%) and G2each site is a pie chart showing the relative prevalence of rotavirusta (available for 2010 only) is included with Ottawa. A pie chart to thein the study for all the years of study.Table 1 Rotavirus identifications shown by genotype, year and surveillance siteCity Year Negative G1P[8] G2P[4] G3P[8] G4P[8] G9P[8] G9P[4] MixedVancouver 2010 3 9 3 5 0 0 0 0Edmonton 2007 1 13 4 0 0 0 0 6Saskatoon 2010 2 5 1 2 0 0 1 0Winnipeg 2007 1 3 0 0 0 0 0 02010 1 2 1 1 0 0 0 0Toronto 2010 0 3 0 0 0 0 0 0Ottawa 2007 2 25 1 1 1 0 0 02008 27 16 3 4 0 0 0 12009 4 20 3 0 0 12 0 12010 1 3 1 2 0 0 0 0Quebec City 2007 4 15 1 7 0 0 0 02008 12 7 1 0 0 0 0 02009 1 6 2 5 0 0 0 2Halifax 2007 1 5 1 1 0 1 0 12008 5 29 0 0 0 0 0 02009 12 3 0 17 2 0 0 12010 0 10 0 0 0 0 1 0Total 77 174 22 45 3 13 2 12McDermid et al. BMC Infectious Diseases 2012, 12:306 Page 4 of 9http://www.biomedcentral.com/1471-2334/12/306P[4] (8.5%). At one site (Ottawa) during 2009 there wasa significantly higher incidence of G9P[8] and a lowerproportion of G1P[8] genotypes (Table 1; 95% CI, modi-fied Wald method for proportions).With the P[8] genotyping assay, we found that all 31of the false negatives from the 1T-1 primer were pickedFigure 2 Genotype prevalence by year for all Canadian sites. The fourup as P[8] samples by the 1T-1DCDN primer. In thispanel (which was deliberately weighted with samplesthat were not picked up by the 1T-1 primer), the 1T-1DCDN primer had a sensitivity of 98% and a specificityof 90%, whereas the 1T-1 primer had a sensitivity of only60% and a specificity of 82%. Thus 1T-1DCDN can beseasons of surveillance are represented from left to right.usefully added to the panel for genotype screening tests,and could potentially replace the 1T-1 primer.A phylogenetic tree constructed with full VP7 genesequences from all of the Canadian G9P[8] isolates in-cluding several prototype G9 sequences from differentregions of the world is shown in Figure 3. The tree showsthat the Canadian G9 isolates fall into the lineage IIImajor subcluster, being more similar to G9 sequencesfrom isolates obtained globally than to previouslysequenced minor subcluster lineage III isolates from Asia.DiscussionHere we report the results of the first systematic, multi-site and multi-season rotavirus genotyping study inCanada. The most prevalent genotype found in childrenwith gastroenteritis at all the sites throughout the fourianMcDermid et al. BMC Infectious Diseases 2012, 12:306 Page 5 of 9http://www.biomedcentral.com/1471-2334/12/306Figure 3 Phylogenetic tree of the VP7 genome segments of Canadsequences. The maximum-likelihood tree was bootstrapped 1000 times. Care designated with “RT” and their numbers end with the year of specimenisolates of rotavirus along with selected G9 prototypeanadian G9P[8] strains are represented in the tree. All Canadian strainscollection.McDermid et al. BMC Infectious Diseases 2012, 12:306 Page 6 of 9http://www.biomedcentral.com/1471-2334/12/306years of the study was G1P[8]. Other common genotypeswere G3P[8] and G2P[4]. Until the present study, onlytwo previous Canadian genotyping studies had been car-ried out: these were restricted to G-genotyping only atsingle sites during single seasons [19,21]. Our currentgenotyping data indicate that the available vaccinesshould be serologically well-matched to the prevalentstrains of rotavirus, based on the genotype prevalence atthese sites. Previous single-site studies in Canada during1999 and 2002 also found G1 as the most prevalentrotavirus genotype [19,21]. Although G1, G2, G3 and G4are the main serotypes present in North America andEurope, other serotypes are important causes of rota-virus gastroenteritis worldwide, such as G5 in SouthAmerica and G8 in Africa [44-49]. G9 strains emergedin the 1990s as a cause of a significant proportion ofrotavirus cases which varied regionally from 5 to 90%[44,50-53]. Although we did not sample every site inevery year in the current study, our results provide re-cent data on the occurrence of rotavirus genotypes inCanada, during the period prior to vaccine roll-out. Oursampling rate is equivalent to about 1 per 100,000 popu-lation over the 4 years of the study in Canada, coveringa median of 4 centres. This sampling rate comparesfavourably with other surveillance studies including onecarried out previously in the United States [48]. We cal-culate that our study would have a 95% probability ofdetecting a rotavirus genotype with an incidence of 1.5%per year. Therefore, this level of sampling may miss anyrare genotypes that occur incidentally and which do notspread in the population beyond one to two percent.G9P[8] strains were identified in the current study butwere mostly restricted to single isolates in a few sitesduring 2 seasons (2007 and 2010) rather than fully emer-ging as a prevalent genotype. Recently, G9 genotypeshave also been identified in the United States and com-prised 39% of the rotavirus genotypes indentified inDetroit between 2007 and 2009 [41]. Since G9 was onlysubstantially present in one site and for one season inCanada (Ottawa in 2009), and could therefore be a geno-type that occurs with irregular frequency in Canada, wedecided to investigate the possible origins of thesestrains using phylogenetic analysis. This shows that theCanadian G9 rotavirus strains collected in the presentstudy are part of G9 lineage III (Figure 3). Although upto six lineages of G9 have been described, most virusesfall into three main lineages, I, II and III: a small numberof G9 strains that do not fit into these lineages have alsobeen reported [43]. The G9 genotype emerged world-wide starting in about 1995, and appeared to spreadworldwide during the 2000s [43]. Sequence analysis indi-cated that one particular subcluster of G9 lineage IIIseemed to spread and cause disease throughout much ofthe world [43]. Most of the circulating G9 rotavirusesworldwide, including all of the G9 specimens from ourstudy, are in the major subcluster of lineage III [43].Thus Canada can be added to the growing list of coun-tries where this lineage has been identified. Lineage Iand II G9 rotaviruses are less common and weredetected primarily within the United States, Japan andIndia in the 1980s and 90s, but reports of new lineagescirculating in other countries underline the importanceof continued rotavirus surveillance. [12,46,54-56].Based on sequencing of a 981 bp region of the openreading frame of VP7, the Canadian G9 strains are moresimilar to isolates from Australia, Brazil and Italy thanthe G9 strain found in Detroit designated MI08/USA(Figure 3). Sequencing data from the two Canadian2010 G9 strains also showed that they have between 96and 98.5 percent identity with 2009 Canadian rotavirusstrains. In addition the 2010 strains are most similar tothe 2007 G9P[8] strain from Halifax, rather than the2009 Ottawa strains (data not shown). Although G9 isnot present in either vaccine, there is serological crossprotection amongst G genotypes and amongst differingP genotypes since the majority of these Canadian G9viruses were G9P[8] and the P[8] antigenic componentis present in both vaccines. Given that two G9P[4]strains were collected in 2010, surveillance of genotypedata is warranted to monitor genotype prevalence asthere is the possibility of an increase in the prevalence ofrarer genotypes, or genetic drift leading to immune es-cape [28,44]. For example, emerging strains such as G12have been detected in New York State [57] and in Italy[58] and may spread to Canada in the future.The local fluctuation of genotype prevalence that isfrequently seen from year to year in circulating rotaviruswas also demonstrated by less frequent genotypes suchas G3P[8], that was more prevalent in 2009 andaccounted for 77% of all rotavirus-positive samples fromHalifax. The high prevalence of G3P[8] in Halifax in2009 was preceded and followed by seasons in which noG3P[8] samples were detected at that site.ConclusionsHemi-nested multiplex PCR is a rapid method for geno-typing rotavirus samples, based on highly conservedgenotype-specific regions [32,36,37]. Nevertheless, muta-tions may cause mispairing resulting in an untypablestrain or a mistyping error [39]. Therefore we monitoredthe primer-binding regions of isolates collected duringthe study for genetic differences that could affect sensi-tivity and specificity. The 1T-1DCDN primer was devel-oped to genotype Canadian P[8] samples that havemismatches in the binding region of the previously used1T-1 primer, which failed to genotype many of theCanadian P[8] isolates. The 1T-1DCDN primer was vali-dated for specificity and sensitivity for genotypingMcDermid et al. BMC Infectious Diseases 2012, 12:306 Page 7 of 9http://www.biomedcentral.com/1471-2334/12/306unidentified Canadian P[8] samples as well as those pre-viously identified with 1T-1, and thus may be useful infuture Canadian genotyping studies.The finding of rotavirus in one third of samples whichwere negative by antigen testing or EM in this study alsoillustrates the lower sensitivity of antigen testing forrotavirus. However, antigen detection tests may be lessprone to detecting low level background infections, andare therefore useful for studies to measure the burden ofrotavirus illness, as well as for investigating the effective-ness of vaccine in decreasing all-cause gastrointestinalillness in the younger age groups. Our findings in thisstudy strongly suggest that currently licensed vaccinesare well matched to the rotavirus strains present in re-cent years in Canada, and that continued surveillance iswarranted to monitor the situation after Canadian uni-versal vaccination programs have been fully introduced.Competing interestsThere were no competing interests for the authors of this study.Authors’ contributionsAM: carried out the molecular genetic studies, sequence analysis and draftedthe manuscript. NL: Contributed to conception and study design andprovided critical review of manuscript. EG: Carried out molecular geneticstudies, sequence analysis. Supervised laboratory work and training. Involvedin analysis and interpretation of data. JAB: Contributed to conception andstudy design and provided critical review of manuscript. KM: carried outmolecular genetic studies, sequence analysis. SAH: Contributed toconception and study design and provided critical review of manuscript. LM:Carried out molecular genetic studies, sequence analysis. PD: Contributed toconception and study design and provided critical review of manuscript. JE:Contributed to conception and study design and provided critical review ofmanuscript. WV: Contributed to conception and study design and providedcritical review of manuscript. TFB: Principal investigator; Supervised laboratorywork. Involved in design of laboratory tests and analysis and interpretation ofdata. Contributed to conception and study design. Writing of final versionsof the manuscript. All authors read and approved the final manuscript.AcknowledgementsThe genotyping study was funded by the Public Health Agency of Canadaand the Canadian Biotechnology Strategy Fund: Genomics Initiative forGovernment Laboratories. We thank the Genomics Core at the NationalMicrobiology Laboratory for sequencing and Jon Gentsch and MichaelBowen of the CDC, Atlanta for reference strains. The Canadian ImmunizationMonitoring Program, Active (IMPACT) is a national surveillance initiativemanaged by the Canadian Paediatric Society and conducted by the IMPACTnetwork of pediatric investigators. IMPACT rotavirus surveillance wassupported by a grant from GlaxoSmithKline Inc. and Merck FrosstCanada Ltd.Investigators and centres participating in IMPACT included: R. Morris, MD,Janeway Children's Health & Rehabilitation Centre, St. John's, Newfoundland;S. Halperin MD, IWK Health Center, Halifax, Nova Scotia; P. Déry, MD, CentreMere-Enfant de Quebec, CHUL, Quebec City, Quebec; D. Moore, MD, TheMontreal Children's Hospital, Montreal, Quebec; M. Lebel, MD, Hôpital Ste-Justine, Montreal, Quebec; N. Le Saux, MD, Children's Hospital of EasternOntario, Ottawa, Ontario; D. Tran, MD, The Hospital for Sick Children, Toronto,Ontario; J. Embree, MD, Winnipeg Children's Hospital, Winnipeg, Manitoba; B.Tan, MD, Royal University Hospital, Saskatoon, Saskatchewan; T. Jadavji, MD,Alberta Children's Hospital, Calgary, Alberta; W. Vaudry, MD, StolleryChildren's Hospital, Edmonton, Alberta; and L. Sauvé, MD, BC Children'sHospital, Vancouver, British Columbia.Author details1National Microbiology Laboratory, Winnipeg, Canada. 2Department ofMedical Microbiology, University of Manitoba, Winnipeg, Canada. 3Division ofInfectious Diseases, CHEO Research Institute, Children’s Hospital of EasternOntario, Ottawa, Canada. 4Vaccine Evaluation Centre, BC Children’s Hospital,University of British Columbia, Vancouver, Canada. 5Canadian Center forVaccinology, IWK Health Centre and Dalhousie University, Halifax, Canada.6Centre Mere-Enfant de Quebec, CHUL, and Laval University, Quebec City,Canada. 7Winnipeg Children's Hospital, Winnipeg, Canada. 8Stollery Children’sHospital and University of Alberta, Edmonton, Canada.Received: 4 June 2012 Accepted: 7 November 2012Published: 15 November 2012References1. 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Grassi T, De Donno A, Guido M, Gabutti G, Collaborative Group for theSurveillance of Rotavirus Infection: G-genotyping of rotaviruses and stoolsamples in Salento, Italy. J Prev Med Hyg 2006, 47(4):138–141.doi:10.1186/1471-2334-12-306Cite this article as: McDermid et al.: Molecular characterization ofrotavirus isolates from select Canadian pediatric hospitals. BMC InfectiousDiseases 2012 12:306.Submit your next manuscript to BioMed Centraland take full advantage of: • Convenient online submission• Thorough peer review• No space constraints or color figure charges• Immediate publication on acceptance• Inclusion in PubMed, CAS, Scopus and Google Scholar• Research which is freely available for redistributionMcDermid et al. BMC Infectious Diseases 2012, 12:306 Page 9 of 9http://www.biomedcentral.com/1471-2334/12/306Submit your manuscript at www.biomedcentral.com/submit


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