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multiplex nested rt-pcr for the detection of porcine enteric viruses.porcine epidemic diarrhea virus (pedv), transmissible gastroenteritis virus (tgev), and porcine group a rotavirus (prv-a) are major viruses causing enteric diseases of piglets. a multiplex nested reverse transcription polymerase chain reaction (multiplex nested rt-pcr) was developed for the detection of these viruses in field samples from piglets with diarrhea. a mixture of (1) three external pairs of primers, yielding in the amplification step two different amplicons with sizes of 950 bp and 31 ...201020170679
circulating human group a rotavirus genotypes in malaysia.this study examined the temporal distribution of rotavirus genotypes in malaysia. rotaviruses from children with diarrhea admitted to hospitals in 1996 (n = 93) and 2007 (n = 12) in two different regions of peninsular (west) malaysia were analyzed for their g and p genotypes using a hemi-nested rt-pcr assay. in the 2007 samples, the dominant strain was g9p[8]. it was identified in 42% of the samples. different strains all possessing the g1 genotype were identified in the rest of the samples. in ...201020166178
genetic variation of g4p[6] rotaviruses: evidence for novel strains circulating between the hospital and community.one hundred forty-six fecal specimens collected between 2007 and 2008 from infants with acute gastroenteritis were screened for rotavirus by elisa with vp6-specific antibody. one hundred twenty-three of the samples (84.2%) were confirmed to be positive for group a rotavirus (community-acquired, n = 90 [73.2%] and nosocomial, n = 33 [26.8%]), and were typed subsequently using rt-pcr and sequence analysis methods. determination of g- and p-type combinations showed that g4p[6] (78.9%) was the most ...201020166174
full genomic analysis and possible origin of a porcine g12 rotavirus strain ru172.human group a rotavirus (gar) g12 strains are regarded as potentially important pathogens for acute gastroenteritis. on the other hand, to date, the only report of detection of g12 in animals was that of a porcine g12p[7] strain ru172. strain ru172 formed a separate g12 lineage, distinct from human g12 strains, and by analyses of deduced amino acid sequences, had a vp4, vp6, nsp4-5 of porcine origin. in the present study, we determined the full-length nucleotide sequences of vp1, vp3, and nsp1-3 ...201020157771
phylogenetic analysis of human p[8]g9 rotavirus strains circulating in brazil reveals the presence of a novel genetic variant.group a rotavirus (rv-a) genotype p[8]g9 has emerged as one of the leading causes of gastroenteritis in children worldwide and currently is recognized as one of the five most common genotypes detected in humans. high intragenotype diversity in g9 rv-a has been observed, and nowadays, based on the genetic variability of the vp7 gene, six different phylogenetic lineages and eleven sublineages were described.201020133185
characterization of group a rotavirus infections in adolescents and adults from pune, india: 1993-1996 and 2004-2007.a total of 1,591 fecal specimens were collected in 1993-1996 and 2004-2007 from adolescents and adults with acute gastroenteritis in pune, india for detection and characterization of rotavirus. at the two time points, group a rotavirus was detected in 8.6% and 16.2% of the adolescents and 5.2% and 17.2% of the adults, respectively. reverse transcription-pcr with consensus primers followed by multiplex genotyping pcr detected common strains g1p[8], g2p[4], g3p[8], and g4p[8] in a total of 53.1% o ...201020087938
molecular characterization of group a human rotavirus among hospitalized children and adults in bangladesh: finding of emerging g12 strain.between july 2004 and june 2006, a total of 1438 fecal samples were collected from hospitalized children and adults with diarrhea at an infectious disease hospital (sk hospital) in mymensingh, bangladesh. the diversity of rotavirus strains were investigated using electropherotyping and reverse transcription-pcr amplification of the vp7 and vp4 genes. group a rotavirus was detected in 171 of 1438(11.9%) specimens: 13.3%(72/543) from children 6 months to </=15 years of age and 11.0%(99/895) from i ...201020046166
comparing electron microscopy and a competitive blocking elisa in the detection of rotaviruses in porcine faeces.analysis of 476 faecal samples from diarrhoeic piglets was performed using electron microscopy (em) and a competitive blocking (cb)-elisa based on monoclonal antibodies to the vp6 protein of group a rotavirus. rotavirus was detected by em and/or cb-elisa in 111 (23.3%) samples. of these, all groups of rotavirus were identified in 83 (74.8%) samples by em (em+), while group a rotavirus was identified in 90 (81.1%) samples by cb-elisa (elisa+). however, only 62 (55.9%) of samples were positive usi ...201120036588
analysis of rotavirus non-structural protein nsp5 by mass spectrometry reveals a complex phosphorylation pattern.genomic replication and partial assembly of rotavirus takes place in cytoplasmic viral structures called viroplasms. nsp5 is a viral phosphoprotein localized in viroplasms and its expression is imperative for viral cycle progress. during infection three isoforms of nsp5 can be observed by sds-page (26, 28 and 33-35kda) and previous reports suggested that they differ in their phosphorylation patterns. in this study we obtained nsp5 from infected cells and by mass spectrometry we were able to iden ...201020036292
viral load and genotypes of noroviruses in symptomatic and asymptomatic children in southeastern brazil.noroviruses (novs) are a major etiological agent of sporadic acute gastroenteritis worldwide.201020004146
[nsp5/nsp6 of rotavirus strain tb-chen and preliminary genotyping analysis].strain tb-chen is a group a rotavirus (rv) isolated from a chinese infant suffering from gastroenteritis in hospital. the nsp5 and nsp6 of strain tb-chen are encoded by the 10th gene segment (816bp in whole length) of the viral genome. the results obtained in this study showed that the nsp5 was encoded in the first open-reading-frame (orf) of the gene segment (from 22bp to 624bp), and nsp6 was encoded in the second orf (from 80bp to 355bp). the nsp5 protein consisted of 200 amino acid residues w ...200919954111
molecular characterization of the vp1, vp2, vp4, vp6, nsp1 and nsp2 genes of bovine group b rotaviruses: identification of a novel vp4 genotype.studies on bovine group b rotaviruses (gbrs) are limited. to date, only the vp6 gene of a single bovine gbr strain and the vp7 and nsp5 genes of a few bovine gbr strains have been sequenced and analyzed. in the present study, using a single-primer amplification method, we have determined the full-length nucleotide sequences of the vp1, vp2, vp4, vp6, nsp1 and nsp2 genes of three bovine gbr strains from eastern india. in all six of these genes, the bovine gbr strains shared high genetic relatedne ...201019936611
rotac: a web-based tool for the complete genome classification of group a rotaviruses.group a rotaviruses are the most common cause of severe diarrhea in infants and children worldwide and continue to have a major global impact on childhood morbidity and mortality. in recent years, considerable research efforts have been devoted to the development of two new live, orally administered vaccines. although both vaccines have proven to confer a good protection against severe rotavirus gastroenteritis, these vaccines will have to be screened and may have to be updated regularly to refl ...200919930627
[evolution of group a rotavirus strains circulating in tunisia over a 3-year period (2005-2007).]background: rotaviruses are the most frequent agents associated with diarrhoea in children worldwide. analysis of mobility of the 11 segments of genomic rna by polyacrylamide gel electrophoresis (page) yields a pattern which is characteristic for a particular rotavirus isolate. the group a rotaviruses can be further characterized by analysis of vp7 and vp4 genes specificities, responsible for rotavirus classification into g and p genotypes, respectively. the aim of the present study was to deter ...200919896286
culturing, storage, and quantification of rotaviruses.group a rotaviruses (rvs) infect the young of numerous animal species and cause acute gastroenteritis. cultivation of animal and human rvs in cells requires proteolytic activation of the viral attachment protein using trypsin. continuous cell lines, such as rhesus monkey kidney cells, as well as primary monkey kidney cells, are routinely used for the growth and characterization of rvs. isolation and cultivation of human rvs from clinical fecal specimens is difficult and adaptation to growth in v ...200919885940
relationship among epidemiological parameters of six childhood infections in a non-immunized brazilian community.epidemiological parameters, such as age-dependent force of infection and average age at infection (<img src='../img/13s1.gif' width=9 height=12>) were estimated for rubella, varicella, rotavirus a, respiratory syncytial virus, hepatitis a and parvovirus b19 infections for a non-immunized brazilian community, using the same sera samples. the for the aforementioned diseases were 8.45 years (yr) [95% ci: (7.23, 9.48) yr], 3.90 yr [95% ci: (3.51, 4.28) yr], 1.03 yr [95% ci: (0.96, 1.09) yr], 1.58 yr ...200919876563
distribution of g (vp7) and p (vp4) genotypes of group a bovine rotaviruses from turkish calves with diarrhea, 1997-2008.group a rotaviruses are major enteric pathogens of calves. in order to investigate the genetic diversity of bovine rotaviruses (brvs), a collection of 53 brvs, detected from diarrheic calves from several turkish geographical areas, between 1997 and 2008 was analyzed by rt-pcr for specificities of the outer capsid proteins vp7 (g type) and vp4 (p type), for the first time. overall, g6 was the predominant g type, detected in 40/53 samples (75.4%), while p[11] was the predominant p type, detected i ...201019854003
[sequence and evolutionary analysis of vp1 gene of ovine rotavirus nt].the ovine rotavirus strain nt isolated from diarrhea lamb in china was considered as a promising vaccine strain. based on the vp1 gene was one of the important structural proteins of rotavirus, we studied on the evolutional characteristics of vp1.200919835167
rotavirus surveillance in europe, 2005-2008: web-enabled reporting and real-time analysis of genotyping and epidemiological data.the first european rotavirus surveillance network, eurorotanet, comprising 16 laboratories in 15 european countries, has been established.200919821712
hospital-based surveillance of rotavirus and other viral agents of diarrhea in children and adults in russia, 2005-2007.during a 2-year period in 2005-2007, we conducted surveillance of group a rotaviruses and other enteric agents among patients hospitalized with acute gastroenteritis in 8 different cities of the russian federation. fecal specimens were gathered from 3208 children (including 2848 children aged <5 years) and 1354 adults who were admitted to hospitals in moscow, st. petersburg, chelyabinsk, nizhnii novgorod, tyumen, khabarovsk, makhachkala, and yakutsk. polymerase chain reaction was performed to de ...200919817602
hospital-based surveillance of rotavirus diarrhea in the people's republic of china, august 2003-july 2007.rotaviruses cause acute diarrhea worldwide. previous studies of rotavirus diarrhea in china found that rotavirus infection is the most common cause of severe diarrhea in young children. in the present study, surveillance of rotavirus diarrhea was conducted involving 9549 children aged <5 years who were admitted for treatment of diarrhea at 11 sentinel hospitals in china from august 2003 through july 2007. group a rotavirus was detected in 3749 (47.8%) of the 7846 fecal specimens by using enzyme- ...200919817597
prevalence of rotavirus in acute diarrhea and its association with clinical signs and symptoms.rotaviruses are the main etiological agents of acute infectious diarrhea in children. thus, the objective of this study is to contribute to the information about the rotavirus variants circulating in brazil. a cross-sectional study was conducted on 124 fecal samples collected from children up to the age of 5 years, with acute gastroenteritis. thirty-one samples were positive for rotavirus a. regarding the g genotyping, 16 (80%) were classified as g1 and regarding p genotyping all strains were cl ...201019793895
human infection with g12 rotaviruses, germany.rotavirus group a g12 genotypes were detected in 3 (1.5%) of 198 stool samples positive for human rotavirus. g12p[6] was present in 2 samples, and a mixed g3g12p[8] was found in 1 sample. phylogenetic analysis of complete open reading frames of all 11 genomic rna segments proved their wa-like genogroup affiliation.200919788829
zoonotic aspects of rotaviruses.rotaviruses are important enteric pathogens of humans and animals. group a rotaviruses (garvs) account for up to 1 million children deaths each year, chiefly in developing countries and human vaccines are now available in many countries. rotavirus-associated enteritis is a major problem in livestock animals, notably in young calves and piglets. early in the epidemiological garv studies in humans, either sporadic cases or epidemics by atypical, animal-like garv strains were described. complete ge ...201019781872
enteric calicivirus and rotavirus infections in domestic pigs.we report the prevalence of rotavirus and calicivirus infections, along with their respective association with diarrhoea in the porcine population of the region of northern spain. a total of 221 samples were collected at random from different farms in the region and from the main slaughterhouse facility in the city of zaragoza. faecal samples were scored as diarrhoeic or normal and grouped into five groups to match general farm management and age criteria: group i (suckling 0-4 weeks), group ii ...201019781118
prevalence of diarrhea viruses in hospitalized children in hong kong in 2008.a total of 209 stool samples were collected from pediatric patients admitted for acute gastroenteritis in a hospital in hong kong, during an 8-month period from january to august 2008, and were tested for the presence of rotavirus, norovirus, sapovirus, adenovirus, and astrovirus using a multiplex rt-pcr assay. the most common virus was rotavirus group a (59 of 209, 28%, mainly serotypes g1, g2, g3, and g9), followed by norovirus group ii (48 of 209, 23%), adenovirus (7 of 209, 3%, serotypes 2, ...200919774686
[rapid detection of norwalk gii with reverse transcription loop-mediated isothermal amplification].to develop a reverse transcription-loop-mediated isothermal amplification (rt-lamp) assay for rapid and sensitive detection of norwalk gii. 4 primers which recognized 6 distinct regions on the rna-dependent rna polymerase gene of norwalk gii were designed and used for lamp assay. norwalk gii rna was amplified under isothermal conditions (65 degrees c) for 120 min, and lamp results were then judged with naked eye, sybr green i staining, electrophoretic analysis and restriction digestion. to evalu ...200919769163
enteropathogens and risk factors for diarrhea in norwegian dairy calves.the aims of the current study were to estimate the prevalence of enteropathogens in calves in norwegian dairy herds, evaluate the clinical consequences of protozoal infections, and identify risk factors for diarrhea. the 135 participating herds were randomly selected from those in the norwegian dairy herd recording system that had at least 15 cow-years. each herd was followed for 1 yr. fecal samples from calves with (n = 68) or without (n = 691) diarrhea were analyzed for the presence of cryptos ...200919762824
in vitro antiherpetic and antirotaviral activities of a sulfate prepared from mimosa scabrella galactomannan.a chemically sulfated galactomannan (brs) from seeds of mimosa scabrella had in vitro antiviral activity against herpes simplex virus 1 (hsv-1), but not against simian rotavirus a/sa11 (sirv-a/sa11). it was examined by (13)c nmr spectroscopy, which showed the sulfate groups to be mainly at c-6 of galactose residues. brs had a selective inhibition against hsv-1 during its attachment step, having an ic(50) lower than 2.5microg/ml, determined by plaque reduction, and a selectivity index of greater ...200919751759
viral gastroenteritis and diversity of rotavirus strains in colombian children: a systematic review.to determine the epidemiological profile of viruses associated with acute gastroenteritis in children younger than 5 years of age and the distribution of rotavirus genotypes circulating in colombia.200819738332
severity of acute gastroenteritis in infants infected by g1 or g9 rotaviruses.group a rotaviruses are the main viral causative agent of acute diarrhea, and cause considerable morbidity in children. g9 rotaviruses have recently emerged all over the world and are thought to give more severe symptoms because of a lack of previous exposure and the absence of maternal antibodies in patients.200919726224
evolutionary consequences of g9 rotaviruses circulating in thailand.infection with group a rotaviruses is the main cause of acute gastroenteritis in infants and young children worldwide. rotavirus g9 is recognized as the fifth most predominant g genotype that spreads throughout the world. in this study, we describe the changing distribution of rotavirus g9 genotype in chiang mai, thailand, from 1989 to 2005. molecular genetic evolutionary analysis of the g9 rotavirus vp7 gene was performed. the g9 rotavirus in thailand was first detected in chiang mai in 1989 wi ...200919716437
detection and characterization of group a rotaviruses in children hospitalized with acute gastroenteritis in norway, 2006-2008.to assess the genetic diversity of rotavirus strains in norway, the distribution of rotavirus genotypes was studied in children admitted to hospital with acute gastroenteritis. the detection of rotavirus in stool samples was compared using an enzyme-linked immunosorbent assay (elisa), an immunochromatographic test and rt-pcr. children <5 years of age admitted to hospital with diarrhea in three large hospitals were enrolled prospectively from march 2006 to february 2008. rotavirus was detected in ...200919697411
environmental dissemination of group a rotavirus: p-type, g-type and subgroup characterization.rotaviruses a (rv-a) infection is the most common cause of acute diarrheal diseases in infants and the dissemination of these viruses in the environment represents a public health hazard. the present study aims to evaluate reverse transcription-polymerase chain reaction (rt-pcr) based protocols for the detection of rv-a genes in different types of environmental samples. rv-a were concentrated by the adsorption-elution method using negatively charged membranes associated with a centriprep concent ...200919657158
national rotavirus surveillance in argentina: high incidence of g9p[8] strains and detection of g4p[6] strains with porcine characteristics.group a rotaviruses are the most frequently detected viral agents associated with diarrhea in infants and children worldwide. it has been estimated that every year almost 120,000 cases of diarrhea associated with rotavirus occur in children under 5 years old in argentina. in this work, we present the rotavirus strain diversity detected during the first 2 years of the national surveillance network for diarrheas implemented by the ministry of health in argentina. during 2006 and 2007 a total of 46 ...200919631763
detection of rotavirus rna and antigens in serum and cerebrospinal fluid samples from diarrheic children with seizures.group a rotavirus (garv) genes (the vp7 and nsp3 genes) in acute-phase cerebrospinal fluid (csf), sera and stool samples from 6 children with convulsions accompanied by garv gastroenteritis were investigated by reverse transcription-polymerase chain reaction (rt-pcr). when the vp7 gene was amplified from the samples, the g genotype (g type) of garv was determined by rt-pcr. garv genes were detected in the csf samples of all 6 children, in 2 of the 3 blood samples, and in all of 4 stool samples. ...200919628905
epidemiology of rotavirus and astrovirus infections in children in northwestern nigeria.recent estimates attribute 527,000 deaths in children less than five years of age to rotavirus diarrhea annually, with 145,000 occurring in sub-saharan africa. human astroviruses have been identified as one of the most frequent causes of infantile diarrhea, second in incidence only to rotavirus. this study was conducted to determine the prevalence of rotavirus and astrovirus and also to establish the circulating strains of rotavirus in a community in nigeria where most diarrheic patients do not ...200819623918
electropherotypes and subgroups of group a rotaviruses circulating among diarrhoeic children in kano, nigeria.it is estimated that about 600,000 children die annually as a result of severe dehydrating diarrhea caused by rotaviruses. the virus is a double stranded rna (dsrna) virus with 11 segments. group a rotaviruses show a characteristic 4-2-3-2 pattern following electrophoresis. the vp6 subgroups, i and ii exist. this work was carried out to study the prevalence of rotavirus infection among children 0-5 years with diarrhea in kano, and to determine the circulating subgroups and electropherotypes and ...200819623917
prevalence of vp7 and vp4 genotypes of human group a rotavirus in infants and children with acute diarrhea in a northern city of bangladesh.the objective of this cross sectional prospective study was to determine rotavirus etiology and prevalence of the different rotavirus genotypes causing acute gastroenteritis in infants and children (</=5years) in mymensingh medical college hospital, bangladesh. the study was conducted during the period from january 2005 to june 2006. a total of 962 stool specimens from diarrheic children were analyzed by polyacrylamide gel electrophoresis (page). rotavirus detection rate was 31.3% (301/962). amo ...200919623146
rotavirus infections in galapagos sea lions.group a rotaviruses infect and cause diarrhea in the young of a broad range of terrestrial mammals, but it is unknown, to our knowledge, whether they infect marine mammals. during february and march of 2002 and 2003, we collected 125 serum samples and 18 rectal swab samples from galapagos sea lion pups (gsl, zalophus wollebaeki), and 22 serum samples from galapagos fur seal pups (gfs, arctocephalus galapagoensis) from nine islands of the galapagos archipelago, ecuador. sera were tested for antib ...200919617482
molecular analysis of g3 rotavirus among infants and children in dhaka city, bangladesh after 1993.between 2004 and 2005, 917 fecal specimens were collected from children below age 5 who presented to child health institute for treatment of diarrhoea in dhaka city, bangladesh. the specimens were screened by rt-pcr for the presence of group a rotavirus and positive stools genotyped. group a rotavirus was detected in 307 stools and serotype g3p[8] strains were detected in nine specimens. sequence analysis clustered the g3 strains into one distinct lineage (lineage i) with other asian g3 strains. ...200919560559
genotypification of bovine group a rotavirus in méxico.bovine scours, frequently provoked by rotavirus infection, causes significant economic losses. nevertheless, no information exists about the bovine rotavirus genotypes present in mexico. this information is necessary for designing efficient vaccines. in this work, 128 samples from diarrheic calves were collected between 2005 and 2006 in 26 dairy and/or beef cattle herds located in 10 regions of mexico, and analyzed for the presence of group a rotavirus. g and p genotypes were determined by pcr i ...200919559115
[rapid detection of rotavirus in water samples using immunomagnetic separation combined with real time pcr].a quantitative and rapid detection method for rotavirus in water samples was developed, by using immunomagnetic separation combined with reverse transcription and real time polymerase chain reaction (ims-rt-real time pcr). magnetic beads coated with antibodies directed against group a rotavirus were used to capture and purify the virus in water samples. the experimental results showed that ims was optimized when 1 ml samples were supplemented with 10 microl of immunomagnetic beads, 2.5 microl of ...200919558103
[norovirus infection in general hospitals for adults].to determine the role of noroviruses in the rise of hospital outbreaks of acute intestinal infections (aii), to study clinical and epidemiological aspects of hospital noroviral infection.200919514424
relationship between electropherotypes and vp7/vp4 genotypes of group a rotaviruses detected between 2000 and 2007 in tunisian children.background: rotaviruses are the most frequent agents associated with diarrhoea in children worldwide. analysis of mobility of the 11 segments of genomic rna by polyacrylamide gel electrophoresis (page) yields a pattern which is characteristic for a particular rotavirus isolate. the group a rotaviruses can be further characterized by analysis of vp7 and vp4 genes specificities, responsible for rotavirus classification into g and p genotypes, respectively. the aim of the present study was to detec ...200919481882
multiplex pcr and multiplex rt-pcr for inclusive detection of major swine dna and rna viruses in pigs with multiple infections.multiplex pcr and multiplex rt-pcr were developed to identify nine viruses in pigs with multiple infections. these viruses are: porcine circovirus type 2 (pcv2), suid herpesvirus 1, porcine parvovirus (ppv), porcine reproductive and respiratory syndrome virus (prrsv), japanese encephalitis virus, porcine rotavirus a (porv-a), porcine epidemic diarrhea virus (pedv), transmissible gastroenteritis virus (tgev), and getah virus. these methods were shown to be high specificity and sensitivity. in the ...200919467264
molecular epidemiology of rotaviruses among healthy calves in japan: isolation of a novel bovine rotavirus bearing new p and g genotypes.a total of 171 fecal specimens collected from healthy calves on a beef farm in gifu prefecture, japan in 2006-2007 were examined for group a rotaviruses by rt-semi-nested pcr targeting the coding region for vp8*. nine specimens were positive for rotavirus. g and p genotyping indicated that one strain was g10p[11]-like and six strains were considered to be the same unknown g and p genotypes. among these six untypeable strains, one strain, azuk-1, was adapted to cell culture and analyzed. sequence ...200919464329
genomic diversity of group a rotavirus strains in patients aged 1-36 months admitted for acute watery diarrhoea in northern india: a hospital-based study.the diversity and capacity of human rotaviruses for rapid evolution and genetic reassortment suggests that rotavirus vaccine should be designed to provide heterotypic protection. the objective of the present study was to provide information on the circulating genotypes of rotavirus and allied baseline epidemiology in lucknow, india. in a cross-sectional study, the prevalence of rotavirus g-p types was studied in patients aged 1-36 months with acute watery diarrhoea. various sociodemographic, env ...201019456832
detection of group c rotavirus in juvenile ferrets (mustela putorius furo) with diarrhea by reverse transcription polymerase chain reaction: sequencing and analysis of the complete coding region of the vp6 gene.nine juvenile ferrets (mustela putorius furo) with a history of diarrhea were severely dehydrated and had distended abdomens and thin-walled small intestines that contained gas and fluid. histologically, small intestines exhibited acute superficial atrophic enteritis. transmission electron microscopy of the small intestine showed rotavirus-like particles within apical vacuoles. reverse transcription polymerase chain reaction (rt-pcr) was negative for group a rotavirus. a group c rotavirus-specif ...200919429992
detection of caliciviruses associated with acute infantile gastroenteritis in salvador, an urban center in northeast brazil.acute gastroenteritis caused by viruses is one of the leading causes of infantile morbidity. the aim of the present study was to investigate the presence of human caliciviruses of the genera norovirus and sapovirus in children up to 3 years of age with acute gastroenteritis from low-income communities in the city of salvador, brazil. this study is an extension of previous work carried out to establish the profile of the most prevalent enteric pathogens present in these communities. in this repor ...200919377793
a study on the possibility of zoonotic infection in rotaviral diarrhoea among calves and buffalo calves in and around kolkata, india.group a and group b rotaviruses are important diarrhea causing agents among calves and buffalo calves. epidemiological studies in indian calves revealed the predominance of group a rotavirus strains with g6, g8, and g10 specificity and group b rotaviruses. a total of 95 fecal samples were collected from calves and buffalo calves affected with diarrhea from an unorganized cattle farm and two cattle markets in and around kolkata, in the state of west bengal of eastern india. rotaviruses were detec ...200919364081
detection of human viruses in rivers of a densly-populated area in germany using a virus adsorption elution method optimized for pcr analyses.transmission of viruses via surface water is a major public health concern. to determine the viral concentration in rivers of a densely-populated area in germany, the virus adsorption elution (viradel) method was optimized for downstream pcr applications. using a high-salt alkaline phosphate buffer for elution, the median recovery efficiency from spiked 1l water samples ranged from 21.3% to 100% for jc polyomavirus, human adenovirus type 5, echovirus 11, and norovirus genogroup i. analyses of 41 ...200919361832
full genomic analysis of a human group a rotavirus g9p[6] strain from eastern india provides evidence for porcine-to-human interspecies transmission.deduced amino acid sequence and phylogenetic analyses of a group a rotavirus g9p[6] strain (designated as mcs/13-07), detected from a 3-year-old child in eastern india, revealed a vp8* closely related to porcine p[6] strains (p[6] sublineage 1d), and the vp7 clustered with g9 lineage-iii strains. to our knowledge, this is the first report of human p[6] strain clustering in sublineage id. thus, to further characterize the evolutionary diversity of strain mcs/13-07, all gene segments were analyzed ...200919333549
first molecular detection of group a rotaviruses in drinking water sources in beijing, china.the most prevalent group a rotavirus found in the diarrheic children was also determined in drinking water sources including raw water, treated water and tap water in beijing, and then the possible contamination contributions to tap water for human consumption were discussed in this study. a total of 26 raw water samples, 77 treated water samples and 143 tap water samples in beijing were collected for analysis of group a rotavirus from april 2006 to august 2007. according to the results, it was ...200919330275
evidence of interspecies transmission and reassortment among avian group a rotaviruses.avian rotaviruses are broadly distributed among birds, but only scarcely characterized on the molecular level. the vp4-, vp6-, vp7- and nsp5-encoding sequences of eight group a rotaviruses from chickens and turkeys determined here indicate a low degree of sequence similarity with mammalian rotaviruses. an nsp6-encoding region was missing in all chicken isolates except for isolate ch2. four novel genotypes (p[30], p[31], g22 and h8) were assigned by the rotavirus classification working group. gen ...200919249805
the first complete genome sequence of a chicken group a rotavirus indicates independent evolution of mammalian and avian strains.group a rotaviruses are a leading cause of gastroenteritis in humans and animals. transmission between mammalian species and humans has been demonstrated repeatedly. here, the first entire genome sequence (19,064 bp) of a chicken rotavirus, strain ch-02v0002g3, is presented. a low degree of nucleotide sequence identity with the mammalian group a rotaviruses is evident for all 11 genome segments, whereas a closer relationship to available rotavirus sequences from avian species has been determined ...200919246068
assignment of the group a rotavirus nsp4 gene into genotypes using a hemi-nested multiplex pcr assay: a rapid and reproducible assay for strain surveillance studies.the rotavirus non-structural protein nsp4 has been implicated in a number of biological functions during the rotavirus cellular cycle and pathogenesis, and has been addressed as a target for vaccine development. the nsp4 gene has been classified into six genotypes (a-f). a semi-nested triplex pcr was developed for genotyping the major human nsp4 genotypes (a-c), which are common in human rotavirus strains but are also shared among most mammalian rotavirus strains. a total of 192 previously chara ...200919208878
viral agents associated with acute gastroenteritis in children hospitalized with diarrhea in lanzhou, china.gastroenteritis is a major cause of childhood morbidity and mortality worldwide. rotavirus, human caliciviruses (hucv), adenovirus, and astrovirus are recognized as common etiologies of acute gastroenteritis.200919201258
diagnosing rotavirus a associated iid: using elisa to identify a cut-off for real time rt-pcr.the use of rt-pcr for diagnosis of group a rotaviruses is increasing, but up to 14% of healthy individuals may be positive by rt-pcr. if rt-pcr is not well correlated with disease, rotavirus a may not always be the cause of illness in rt-pcr positive patients with infectious intestinal disease (iid).200919179107
[nucleotide sequence analysis of the nsp4 gene from group a rotaviruses isolated in nizhni novgorod].the complete nucleotide sequences of the nsp4 gene from russian isolates of group a rotaviruses with different g[p] types obtained in nizhni novgorod have been first ascertained. they belong to their affiliation to three nsp4 (a, b, and c) of the 5 known genotypes. eleven strains had nsp4 genotype. phylogenetic analysis based on the complete nucleotide sequences of the nsp4 gene revealed the sequence homology of g1p[8] viruses with the strains circulating in scandinavia and that of the g3 type v ...200819172905
seasonal shifts of group a rotavirus strains as a possible mechanism of persistence in the human population.this article demonstrates how the seasonal predominance of a new rotavirus strain in asuncion, paraguay is correlated with a wide spectrum of age groups of children infected in that given season. therefore, this study provides new evidence to support the idea that seasonal shift of rotavirus strains is a possible mechanism used by the virus to evade herd immunity (acquired by the population due to previous infections) and, thus, ultimately persist in that population.200919152398
[expression and immunogenicity analysis of recombinant plasmid pw425et-vp7 of porcine rotavirus a in lactobacillus].we tested the antigenicity of porcine rotavirus vp7 gene expressed in lactobacillus.200819149168
group a rotavirus genotypes and the ongoing brazilian experience: a review.brazil was the first latin american country to introduce universal group a rotavirus (rv-a) vaccination in march 2006, resulting in a unique epidemiological scenario. since rv-a first identification in brazil, 2,691 rv-a-positive stool samples, collected between 1982- 2007, were typed by independent research groups throughout the country. in the pre-vaccination era, 2,492 rv-a-positive samples collected from 1982-2005 were successfully typed, while 199 samples were analyzed from 2006-2007. accor ...200819148411
intranasal immunization of mice with vp2 dna of human rotavirus a induces cellular and humoral immunity.dna vaccination using a plasmid encoding human rotavirus a (hurv-a) inner capsid vp2 was examined in a mouse model. balb/c mice were immunized intranasally (i.n.) with a vp2 dna vaccine that induced cellular and humoral immune response to hurv-a. the increased levels of cytokines ifn-gamma and il-4 and the production of anti-vp2 igg antibodies were detected. in addition, splenocyte proliferation detected by mtt test was enhanced in the mice treated with the vaccine. these results may encourage t ...200819143478
characterization of novel vp7, vp4, and vp6 genotypes of a previously untypeable group a rotavirus.rotavirus is the most common cause of acute gastroenteritis among infants and young children throughout the world, but rotavirus cases in developing countries account for nearly all of the approximately 600,000 annual deaths. we studied the epidemiology of rotavirus in 22 rural communities in northern coastal ecuador over a five-year period. from 250 rotavirus positive stool specimens, the percentage that could not be rt-pcr genotyped for vp4 and vp7 was 77% and 63%, respectively. the possibilit ...200919131083
phylogenetic analysis of rotaviruses with predominant g3 and emerging g9 genotypes from adults and children in wuhan, china.prevalence and phylogenetic relatedness of rotaviruses causing diarrheal diseases in children and adults were analyzed in wuhan, china. during a period between june 2006 and february 2008, group a rotavirus was identified in 24.9% (280/1126) and 7.6% (83/1088) of specimens taken from children and adults, respectively. g3p[8] was the most frequent genotype in both children (66.3%) and adults (62.7%), followed by g1p[8] (20.3% and 26.2%, respectively). g9 was detected in specimens from six childre ...200919107964
molecular characterization of rotaviruses, noroviruses, sapovirus, and adenoviruses in patients with acute gastroenteritis in thailand.outbreaks of viral gastroenteritis occur worldwide including thailand. unfortunately, there is limited information since etiologic agents have not been identified in several outbreaks of nonbacterial gastroenteritis. the genotype of enteric viruses causing acute gastroenteritis in thailand was determined using reverse transcription-multiplex polymerase chain reaction and dna sequencing. from january 2006 to february 2007, stool samples were collected from patients with acute gastroenteritis of a ...200919107961
analysis of the vp6 gene of human and porcine group a rotavirus strains with unusual subgroup specificities.full-length vp6 amino acid sequences of human and porcine rotaviruses with subgroup (sg) (i + ii) and sg non-(i + ii) were analyzed in comparison with those of sg i and sg ii. in human rotaviruses, the strains in the same sg shared a very high degree of amino acid identity, ranging from 97.4% to 99.4% for sg i, 95.9% to 100% for sg ii, and 99.4% to 100% for sg non-(i + ii), while viruses in different sgs shared somewhat lower sequence identity at 90.4-93.1%. conserved amino acids that distinguis ...200919031468
molecular characterization of vp4, vp6, vp7, nsp4, and nsp5/6 genes identifies an unusual g3p[10] human rotavirus strain.an unusual strain of human rotavirus g3p[10] (cmh079/05) was detected in a stool sample of a 2-year-old child admitted to the hospital with severe diarrhea in chiang mai, thailand. analysis of the vp7 gene sequence revealed highest identities with unusual human rotavirus g3 strain cmh222 at 98.7% on the nucleotide and 99.6% on the amino acid levels. phylogenetic analysis of the vp7 sequence confirmed that the cmh079/05 strain formed a cluster with g3 rotavirus reference strains and showed the cl ...200919031442
[evaluation of two immunochromatography kits for rapid diagnosis of rotavirus infections].a prospective study was conducted to evaluate two immunochromatography (icg) commercial kits for diagnosis of rotavirus infection, vikia rota-adeno (biomérieux) and simple rota-adeno (operon). reverse transcriptase and polymerase chain reaction (rt-pcr) with specific primers for the vp7 gene of group a rotavirus was used as the reference method. the sensitivity and specificity of the icg tests compared with those of the reference method were 98.4% and 84.8%, respectively, for simple rota-adeno ( ...200819024505
supplementing suckling rats with whey protein concentrate modulates the immune response and ameliorates rat rotavirus-induced diarrhea.group a rotaviruses (rv) are the most common causative agents of acute gastroenteritis in children <2 y. the present study was designed to establish the effect of a bovine whey protein concentrate (wpc) in a rv infection model in suckling rats. from d 3 of life, suckling lewis rats received daily supplements of wpc, wpc plus lactoferrin (lf), standard infant formula (sif), or water (rv-infected group and an untreated, uninfected reference group). on d 8 of life, heterologous simian rv sa-11 was ...200819022963
[rotavirus vaccine].rotavirus gastroenteritis is one of the most important diseases in young children. there are 400,000 to 600,000 deaths annually among children under 5 years old in the world, especially in developing countries. rotavirus vaccine has been developed since 1980s. however vaccine development and trial took a long time before the commercial available. one reason is the genetic variability of rotavirus, there are more than 10 serotypes (g types) in group a rotavirus. another reason is side reaction su ...200818939500
unexpected substitution of dominant rotavirus g genotypes in french hospitalized children over five consecutive seasons.the study was designed to evaluate the circulation of group a rotaviruses in french hospitalized children, and to detect unusual strains. this prospective study was conducted from 2001 to 2006 in children consulting for acute diarrhea at the pediatric emergency department in three french university hospitals. the rotaviruses were detected by rapid test and genotyped by rt-pcr on the basis of their outer capsid proteins vp4 (p-type) and vp7 (g-type). the stools from 757 children were analyzed. g1 ...200918855026
use of polymerase chain reaction for the differentiation of group a bovine rotavirus g6, g8, and g10 genotypes in the north island of new zealand.to determine the presence of group a rotavirus g6, g8, and g10 genotypes in calves in the north island of new zealand.200818836501
[rotaviruses in younger children in novosibirsk in 2005-2007: detection and genotyping].examination of 1898 patients with acute enteric infection from march 2005 to february 2007 showed that group a rotaviruses were the most frequent cause (35%) of acute gastroenteritis among children under 3 years of age. majority of cases of rotavirus infection was detected in infants under 1 year of age (71.8%). the peak of sporadic incidence was observed between february and may. high rate of mixed infection (45.6%) was observed - associations of rotaviruses with other viruses (noroviruses, ast ...200818819400
molecular characterization of vp6-encoding gene of group a human rotavirus samples from central west region of brazil.group a rotaviruses are the main cause of acute gastroenteritis in children worldwide. the intermediate capsid protein vp6 encoded by segment 6 of the dsrna genome is the major structural component of the virus and it is highly antigenic and immunogenic. vp6 is responsible for group and subgroup (sg) specificities, allowing classification of group a rotavirus into sg i, sg ii, sg i + ii, and sg non-i-non-ii. vp6-encoding gene of 154 group a human rotavirus samples of different g and p genotypes ...200818814260
emergence of the gii4/2006b variant and recombinant noroviruses in china.noroviruses are an important cause of acute gastroenteritis. increasing data showed that the gii-4 strains are predominant worldwide and new gii-4 variants emerge every 1-2 years causing major epidemics. surveillance of gastroenteritis in hospitalized children under 5 years of age in china is described. among 1,110 specimens, 114 (10.3%) were positive for noroviruses, which was higher than adenoviruses (7.6%), astroviruses (3.5%), and sapoviruses (0.9%) and only lower than group a rotaviruses (4 ...200818814250
an outbreak of group a rotavirus g1p[8] in an elementary school, nara prefecture. 200818806362
a novel genomic constellation (g10p[3]) of group a rotavirus detected from buffalo calves in northern india.group a bovine rotaviruses cause gastroenteritis and calf mortality leading to significant economic losses to dairy farmers in india. due to segmented nature of the rna genome and wide host range, vast genetic and antigenic diversity exists among different isolates of rotavirus. molecular characterization of locally prevalent group a rotavirus strains in buffalo population in north india was undertaken. out of a total of 455 faecal samples, 21 samples (4.61%) were positive for bovine rota virus ...200818793686
human group a rotavirus p[8] hun9-like and rare op354-like strains are circulating among diarrhoeic children in eastern india.during 2004-2006, group a rotavirus p[8] strains were the major vp4 genotype (43.2%, n = 317) among diarrhoeic children in eastern india. phylogenetic analysis of vp8* amino acid sequences of 16 of these strains with other p[8] strains revealed four distinct lineages. p[8] strains from eastern india clustered within rare op354-like and hun9-like lineages, pointing towards co-prevalence of divergent p[8] strains. although it is unclear whether the observed genetic diversity might affect to some e ...200818787756
prospective study of etiologic agents of acute gastroenteritis outbreaks in child care centers.to investigate the etiology of outbreaks of acute gastroenteritis (age) in children attending childcare centers (cccs) in north carolina between october 2005 and march 2007.200918783794
evaluation of rapid assays for the detection of bovine coronavirus, rotavirus a and cryptosporidium parvum in faecal samples of calves.rapid immunochromatographic assays for detecting infections with bovine coronavirus (bcv), rotavirus a and cryptosporidium parvum in calf faeces were evaluated using as gold standards a reverse transcriptase polymerase chain reaction (bcv and rotavirus) and a sedimentation-flotation technique (c. parvum). rapid tests for the detection of bcv and rotavirus showed a high specificity (96.4% and 95.3%, respectively), but a relatively low sensitivity (60.0% and 71.9%, respectively). sensitivity and s ...200918778958
electrophoretic rna genomic profiles of rotavirus strains prevailing among hospitalized children with acute gastroenteritis in tehran, iran.human rotavirus has now been established as the most important cause of childhood gastroenteritis worldwide. the rna genomic electrophoretic diversity of group a rotavirus strains in tehran, iran, during april 2002 through march 2005, was investigated in 1250 stool specimens.200818759520
increase in prevalence of human group a rotavirus g9 strains as an important vp7 genotype among children in eastern india.in addition to four globally important group a rotavirus (garv) vp7 genotypes (g1-g4), recent surveillance studies have revealed importance of g9 strains as an aetiological agent of infantile diarrhoea.200818755625
the post-tsunami outbreak of diarrhoeal diseases in car nicobar island, india, was caused by human group a rotavirus g2 strains. 200818723226
molecular characterization of equine rotavirus in ireland.group a rotaviruses are important causative agents of severe, acute dehydrating diarrhea in foals. a total of 86 rotavirus-positive fecal samples, collected from diarrheic foals from 11 counties in three of the four provinces of ireland, were obtained from the irish equine centre in kildare during a 7-year (1999 to 2005) passive surveillance study and were characterized molecularly to establish the vp7 (g type) and vp4 (p type) antigenic specificities. fifty-eight samples (67.5%) were found to c ...200818716232
vp6 capsid protein of chicken rotavirus strain ch2: sequence, phylogeny and in silico antigenic analyses.the inner capsid protein vp6 of group a rotavirus possesses group and subgroup epitope specificities. avian rotaviruses have a unique vp6 that is antigenically different from its mammalian counterpart. the lack of information on the vp6 protein of chicken rotavirus strain, ch2, at the genetic and antigenic level was a major motivation for this work. sequencing of the complete cdna of the vp6 gene of ch2, revealed a nucleotide (amino acid) identity that varied from 78.3 to 98.5% (86.4-98.2%) when ...200818680770
a survey of specific pathogen-free chicken flocks for antibodies to chicken anaemia agent, avian nephritis virus and group a rotavirus.sera originating from specific pathogen-free flocks from europe, north america and australia were examined by indirect immuno-fluorescence for antibodies to chicken anaemia agent, avian nephritis virus and group a rotavirus. some flocks were free of infection, but others showed evidence of infection with one, two or all of these viruses. in view of the pathogenic potential of these viruses and the possibility that they may be egg-transmitted, it is recommended that their elimination from specifi ...198918679854
molecular characterization of rare bovine group a rotavirus g15p[11] and g15p[21] strains from eastern india: identification of simian sa11-like vp6 genes in g15p[21] strains.during a surveillance study (november 2001-march 2005), one rare g15p[11] and two rare g15p[21] bovine group a rotavirus strains were detected in diarrhoeic calves in eastern india. sequence analysis of the vp8*, vp6, nsp4 and nsp5 genes of the g15p[11] strain confirmed its bovine origin. although the nsp4 and nsp5 genes of the two g15p[21] strains were of bovine origin, their vp6 genes shared higher nucleotide and amino acid identities with simian strain sa11 (92.5-93.1% and 98.5-98.7%) than bo ...200818679786
molecular characterization of vp4, vp6 and vp7 genes of a rare g8p[14] rotavirus strain detected in an infant with gastroenteritis in italy.in this study, the molecular characterization of a rare g8p[14] group a rotavirus (garv) strain detected in northern italy during the 2004-2005 epidemiological rotavirus season is described. two hundred and seventy three rotavirus-like particle positive stools out of 856 stools from children (31.9%) hospitalized with gastroenteritis were analyzed using polyacrilamide gel electrophoresis and 271 garvs were genotyped by vp7 and vp4 specific rt-pcrs. one strain (pr/1300/04) with a long electrophero ...200818674577
determination of human rotavirus vp6 genogroups i and ii by reverse transcription-pcr.based on nucleotide sequence and phylogenetic analysis of the partial vp6 genes, group a rotaviruses can be mainly differentiated into two genogroups. in this study, a method employing reverse transcription-pcr (rt-pcr) and degenerate primers was established to assign the vp6 genogroup. vp6 genogroup i and genogroup ii could be determined according to the sizes of the amplicons: 380 and 780 bp, respectively. the vp6 genogroup of human reference strains of g1 to g4 and g9 types and rotateq vaccin ...200818667595
genomic characterization of a novel group a lamb rotavirus isolated in zaragoza, spain.an ovine rotavirus (ovr) strain, 762, was isolated from a 30-day-old lamb affected with severe gastroenteritis, in zaragoza, spain, and the vp4, vp7, vp6, nsp4, and nsp5/nsp6 genes were subsequently characterized molecularly. strain ovr762 was classified as a p[14] rotavirus, as the vp4 and vp8* trypsin-cleavage product of the vp4 protein revealed the highest amino acid (aa) identity (94% and 97%, respectively) with that of the p11[14] human rotavirus (hrv) strain pa169, isolated in italy. analy ...200818661221
incidence and clinical characteristics of group a rotavirus infections among children admitted to hospital in kilifi, kenya.rotavirus, predominantly of group a, is a major cause of severe diarrhoea worldwide, with the greatest burden falling on young children living in less-developed countries. vaccines directed against this virus have shown promise in recent trials, and are undergoing effectiveness evaluation in sub-saharan africa. in this region limited childhood data are available on the incidence and clinical characteristics of severe group a rotavirus disease. advocacy for vaccine intervention and interpretation ...200818651787
outbreak of rotavirus variant p[8] in seoul, south korea.an epidemiologic study was performed to determine the genetic variability of rotaviruses in seoul, south korea. in 3,174 stool specimens from children with acute diarrhea at five referral hospitals, 571 (18%) possessed the antigen of group a rotavirus detectable by elisa--10.8% in 2004 and 28.1% in 2005. vp7 genotyping revealed that the g3 type was found in 25.6% of all typed isolates, g4 in 23.8%, g2 in 21.6%, and g1 in 17.6%. vp4 genotyping showed that the p[8] type was detected in 66.7%, p[6] ...200818649327
detection and characterization of group c rotaviruses in asymptomatic piglets in ireland.group c rotaviruses are important human enteric pathogens that have also been detected in a variety of mammalian species, including pigs. group c rotaviruses have been identified in piglets with diarrhea, but their ecology remains to be elucidated. by screening of 292 fecal samples collected from 4- to 5-week-old asymptomatic pigs from four herds in ireland between 2005 and 2007, 13 (4.4%) samples tested positive by reverse transcription-pcr for group c rotavirus. group a rotaviruses were also d ...200818632912
llama-derived single-chain antibody fragments directed to rotavirus vp6 protein possess broad neutralizing activity in vitro and confer protection against diarrhea in mice.group a rotavirus is one of the most common causes of severe diarrhea in human infants and newborn animals. rotavirus virions are triple-layered particles. the outer capsid proteins vp4 and vp7 are highly variable and represent the major neutralizing antigens. the inner capsid protein vp6 is conserved among group a rotaviruses, is highly immunogenic, and is the target antigen of most immunodiagnosis tests. llama-derived single-chain antibody fragments (vhh) are the smallest molecules with antige ...200818632867
recommendations for the classification of group a rotaviruses using all 11 genomic rna segments.recently, a classification system was proposed for rotaviruses in which all the 11 genomic rna segments are used (matthijnssens et al. in j virol 82:3204-3219, 2008). based on nucleotide identity cut-off percentages, different genotypes were defined for each genome segment. a nomenclature for the comparison of complete rotavirus genomes was considered in which the notations gx-p[x]-ix-rx-cx-mx-ax-nx-tx-ex-hx are used for the vp7-vp4-vp6-vp1-vp2-vp3-nsp1-nsp2-nsp3-nsp4-nsp5/6 encoding genes, resp ...200818604469
molecular characterization of the nsp4 gene of human group a rotavirus samples from the west central region of brazil.nonstructural protein 4 (nsp4), encoded by group a rotavirus genome segment 10, is a multifunctional protein and the first recognized virus-encoded enterotoxin. the nsp4 gene has been sequenced, and five distinct genetic groups have been described: genotypes a-e. nsp4 genotypes a, b, and c have been detected in humans. in this study, the nsp4-encoding gene of human rotavirus strains of different g and p genotypes collected from children between 1987 and 2003 in three cities of west central regio ...200818592101
human, porcine and bovine rotaviruses in slovenia: evidence of interspecies transmission and genome reassortment.a surveillance of human, porcine and bovine rotaviruses was carried out in slovenia in 2004 and 2005. stool samples were collected from a total of 406 pigs (373 from asymptomatic animals), 132 cattle (126 from asymptomatic animals) and 241 humans (all with diarrhoea), tested for group a rotaviruses using rt-pcr and analysed by sequencing. the aims of the study were to determine the incidence of asymptomatic rotavirus infection in animals, to look for evidence of zoonotic transmission and to dete ...200818559940
rotavirus infection in children and adult patients attending in a tertiary hospital of são paulo, brazil.during the period of january 2003 to december 2005, 3,768 stool samples were received in the microbiology laboratory for rotavirus antigen detection from outpatients and inpatients of albert einstein hospital, sp. fresh stool samples from children and adults were analyzed by two methodologies: during 2003 and 2004 by latex agglutination (slidex rotavirus, biomerieux) and 2005 by an immunochromatographic assay for the combined detection of rotavirus and adenovirus (vikia rota-adeno, biomerieux). ...200818553013
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