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Group A rotaviruses (RVA) are one of the major cause of acute gastroenteritis (AGE) in young children, being responsible for up to 250.000 deaths worldwide, mostly in developing countries. The two outer capsid proteins VP7 (glycoprotein, G-genotype) and VP4 (protease-sensitive protein, P-genotype) are the basis for the binary RVA nomenclature. Although at least 36 G-types and 51 P-types of rotavirus are presently known, most RVA infections in humans, worldwide as well as in Italy, are related to six major G/P combinations: G1P[8], G2P[4], G3P[8], G4P[8], G9P[8] and G12P[8]. In November 2016, in the framework of the Italian 2016/17 rotavirus surveillance season, a total of 22 rotavirus-positive samples from hospitalized children presenting AGE symptoms were collected in a small area of Central Italy (Ancona, Marche). After genotyping, 3 samples presented the G9P[4] genotype. In order to better understand the origin of these uncommon RVA strains causing dehydrating diarrhea in three children, the strains RVA/Human-wt/ITA/AN18/2016/G9P[4], RVA/Human-wt/ITA/AN19/2016/G9P[4] and RVA/Human-wt/ITA/AN22/2016/G9P[4] were subjected to nucleotide sequencing of all the 11 gene segments to define their genomic constellation. Nucleotide sequencing revealed that the genomic constellation of the three strains was G9-P[4]-I2-R2-C2-M2-A2-N2-T2-E2-H2, highlighting human origin for all the gene segments investigated. The molecular characterization of RVAs and the continue monitoring of their circulation is needed to better define the epidemiology of these pathogen and to detect the emergence of viral variants presenting a high spreading potential in humans in the post-vaccination era.

Infection with a rare G3P[19] rotavirus A strain was identified in an immunosuppressed patient in Italy. The strain showed a P[19] viral protein 4 gene and a complete AU-1–like genomic constellation. Phylogenetic analyses showed high nucleotide identity between this strain and G3P[19] rotavirus A strains from Asia, indicating possible reassortment events.

In Tunisia in 2008, an unusual G6P[9] rotavirus, RVA/human-wt/TUN/17237/2008/G6P[9], rarely found in humans, was detected in a child. To determine the origin of this strain, we conducted phylogenetic analyses and found a unique genotype constellation resembling rotaviruses belonging to the feline BA222-like genotype constellation. The strain probably resulted from direct cat-to-human transmission.

The aim of this study was to determine the occurrences of the group A rotavirus (RVA), norovirus (NoV) and human adenovirus (HAdV) in the surface waters of an urban lagoon (Rodrigo de Freitas Lagoon) in the city of Rio de Janeiro, Brazil. During one year of surveillance, water samples were obtained from the lagoon and other interconnected ecosystems (river and beach). The samples were concentrated using an adsorption-elution method with a negatively charged membrane and tested by qualitative and quantitative polymerase chain reaction assays. RVA was the most prevalent virus detected (24.3%) with a viral load ranging from 3.0 x 10¹-5.6 x 10(4) genome copies/L, followed by NoV (18.8%) and HAdV (16.7%). Considering water samples suitable for bathing, according to Escherichia coli criterion (< 2,000 most probable number/100 mL), viruses were detected in 50% (57/114) of them. Physicochemical parameters were also measured and showed possible correlations between turbidity and RVA presence and between pH and NoV presence. These data demonstrate the importance of considering viral parameters to ensure water quality and the utilisation of these parameters as additional tools for the characterisation of environmental contamination.

Diarrhea and deaths in new-born camel calves were noticed by veterinary investigators and pastoralist in Saudi Arabia to be very high. Hence, it is thought to be necessary to investigate this problem from the virological and bacteriological point of view. The role of pathogenic bacteria and viruses in six different towns of North Province (Al-Assafia, Arar, Domat Aljandal, Hail, Skaka and Khoa) in Saudi Arabia was studied. Survey was conducted in diarrheic camel calves aged 12months or younger. In our study calf diarrhea was reported in 184 out of 2308 camels examined clinically during one year, the prevalence of diarrhea was found to be 8.0% in calves ranging from one month to one year. In the present study group A rotavirus and Brucella abortus were detected in 14.7% and 8.98%, respectively, using ELISA technique. Escherichia coli was isolated from diarrheic calf camel (58.2%) 99/170 samples during dry and wet season. Salmonella spp. and Enterococcus spp. were detected in 12% and 8.8% of the specimens, respectively. In this study enterotoxogenic E. coli (ET E. coli) was isolated from 7% of diarrheic camel, which indicates the strong correlation between the camel calf diarrhea and the detection of enterotoxogenic E. coli. This study represented the first report for the detection of group A rotavirus and B. abortus antigen and antibodies in calf camels in Saudi Arabia. It is recommended that the disease should be controlled by vaccination in calf camels.

Summary Background Very little is known about human rotaviruses in the northwest of China. To investigate the genomic diversity, we evaluated the distribution of rotavirus genotypes in this region covering a 10-year period (1996–2005). Methods Rotavirus antigen was detected in stool specimens by enzyme immunoassay (EIA), and G and P genotyping was performed by reverse transcription-polymerase chain reaction and nucleotide sequencing methods. Results A total of 783 stool specimens collected from children with diarrhea, under 5 years of age, attending an urban hospital in Xinjiang were tested for rotavirus antigen, and 398 (50.8%) were positive. Overall, the most prevalent rotavirus genotype was G1P[8] (40.0%), followed by G3P[8] (17.5%), G2P[4] (8.3%), and G2P[6] (6.5%). G1 rotavirus was the most prevalent genotype until 2004. However, in 2005, G3 rotavirus (51.9%) became a dominating strain. Only one G9 strain was isolated in this region (isolated for the first time in 1999) and it became a more prevalent strain (21.2%) in 2005. Conclusions The results of this study are of importance to the decision makers in the evaluation of rotavirus vaccines in China.

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 region of Brazil was characterized. NSP4 gene of 153 rotavirus-positive fecal samples was amplified by reverse transcriptase-polymerase chain reaction and then sequenced. For phylogenetic analysis, NSP4 nucleotide sequences of these samples were compared to nucleotide sequences of reference strains available in GenBank. Two distinct NSP4 genotypes could be identified: 141 (92.2%) sequences clustered with NSP4 genotype B, and 12 sequences (7.8%) clustered with NSP4 genotype A. These results reinforce that further investigations are needed to assess the validity of NSP4 as a suitable target for epidemiologic surveillance of rotavirus infections and vaccine development.

INTRODUCTION: During the period from 2000 to 2002, 79 rotavirus-positive stool samples were collected from children presenting diarrhea in the Western Brazilian Amazon. METHODS: Molecular characterization of the G and P genotypes was performed using RT-PCR and electropherotyping analysis by polyacrylamide gel electrophoresis. RESULTS: A total of 59 samples were confirmed as group A rotavirus. A long electrophoretic profile was exhibited by the G1P[8], G3P[8], and G4P[8] genotypes. The G1P[8] genotype was found in greater proportion. The short electropherotype was exhibited only by G2 genotype strains. CONCLUSIONS: The proportion of the rotavirus genotypes observed was not different from that in other areas of Brazil. This study is the first genotyping of rotavirus in the Western Brazilian Amazon. INTRODUÇÃO: Entre 2000 e 2002, 79 amostras positivas para rotavírus foram coletadas de crianças com diarreia na Amazônia ocidental brasileira. MÉTODOS: Para a caracterização molecular dos genótipos G e P foram realizadas as reações de RT-PCR e a análise dos eletroferotipos por eletroforese em gel de poliacrilamida (PAGE). RESULTADOS: 59 amostras foram confirmadas como pertencentes ao rotavírus grupo A. Os genótipos G1P[8], G3P[8] e G4P[8] apresentaram perfis eletroforéticos longos. O genótipo G1P[8] foi encontrado em maior proporção. O eletroferotipo curto ocorreu apenas em genótipos G2. CONCLUSÕES: A proporção dos genótipos de rotavírus observada não foi diferente de outras áreas do Brasil. Este estudo é a primeira genotipagem de rotavírus na Amazônia ocidental brasileira.

A freqüência de rotavírus do grupo A foi avaliada em 194 amostras, de fezes de bezerros, com idade entre 2 e 60 dias, provenientes de 33 propriedades da bacia leiteira de Pará de Minas, Minas Gerais. Utilizando um sistema de ELISA monoclonal direto, 33 (17%) destas amostras foram positivas para rotavírus do grupo A. Do total de animais positivos para rotavírus do grupo A, 60,6% apresentavam diarréia no momento da coleta. Das 33 propriedades amostradas, a presença de rotavírus foi detectada em 19 (57,6%). Dentre todas as amostras examinadas, somente 7 apresentaram simultaneamente rotavírus e Cryptosporidium spp. The frequency of group A rotaviruses was evaluated in 194 calves, 2- to 60-days old, from 33 farms in Pará de Minas, Minas Gerais State, Brazil. One hundred and ninety four fecal specimens were tested in a direct monoclonal ELISA and 33 (17%) were positive to group A rotavirus. Diarrhea was present in 60,6% of rotavirus-infected calves at the time of feces collection. Group A rotaviruses were detected in 19 (5.,6%) of the studied farms. Among all calves tested only seven were infected simultaneously by group A rotavirus and Cryptosporidium spp.

This series of studies used antigenic and genetic analysis to examine rotavirus strains prevailing in humans and animals in Thailand. The results are summarized as follows: 1） A number of fecal specimens were collected from humans （patients with diarrhea）, cattle （young calves with diarrhea） and pigs （piglets with diarrhea） in Thailand. These were screened for rotavirus by polyacrylamide gel electrophoresis （PAGE） of segmented viral genome and enzyme-linked immunosorbent assay （ELISA） with group A-specific monoclonal antibodies. Group A rotaviruspositive specimens were then examined for their subgroup and G serotype specificity using subgroup （I-and II-） specific monoclonal antibodies and G serotype （1-, 2-, 3-, 4- and occasionally 6-） specific monoclonal antibodies, respectively. G serotype specificity of the specimens untypable by ELISA was then examined by polymerase chain reaction （PCR） using serotype-specific primers, which showed very high sensitivity in serotype determination. 2） The rate of rotavirus detection in pediatric diarrhea was calculated at 36.9% on the basis of the examination of diarrhea specimens collected in Bangkok in 1991-94. Rotavirus gastroenteritis was shown to occur most frequently in infants and young children equal to or less than 2 years of age with a peak at 6-11 months of age. The male to female ratio was calculated at 1.36. Monthly distribution of the occurrence of rotavirus gastroenteritis in Bangkok indicated that the illness tends to cluster in September through January. 3） Although as a whole G serotype 1 was the most often detected in this series, the frequency of detection of individual serotypes differed greatly by year. While in the 1983-84 survey G serotype 4 was the most frequent, both G serotypes 1 and 2 dominated in the 1987-88 survey. In the following two years of 1988-89 and 1989-90, G serotype 1 predominated, while in 1990-91 survey G serotype 3, which had been detected only rarely until then, surpassed other G serotypes 1 and 2. G serotype 1 was the most frequent in 1991-92, second to G serotype 2 in 1992-93 and again predominant in 1993-94. 4） It was unexpected that all bovine rotavirus strains found in the present studies were antigenically non-G serotype 6, since most bovine rotaviruses reported until then belonged to G serotype 6. The RNA patterns of these strains were grouped into the three electropherotypes a, b and c. Sequence analysis of the VP7 gene of strain 61A （representing the electropherotype c） indicated that it does not belong to G serotype 1, 2, 3, 4, 5, 6, 8, 9, or 11. These results formed the basis of our subsequent antigenic and genetic studies showing that while the G serotype of strains A44 （electropherotype a） and 61A is 10, that of strain A5 （electropherotype b） is 8, which is discussed in this paper. ?5） Serotyping by ELISA showed that the majority of porcine rotavirus strains （7/13） detected in 1987 was G serotype 3. Twenty-three group A rotavirus strains detected in 1990-91 were grouped by RNA PAGE into six electropherotypes a-f. The G serotypes assigned to these by ELISA and PCR were as follows: G serotype 3 （three strains of electropherotypes a and c）, G serotype 10 （14 strains of electropherotype d） and undetemined serotype （6 strains）. The presence of G serotype 10 porcine rotavirus, a finding unreported to date, has been further confirmed by serologic and genetic analysis in this laboratory （Pongsuwanna et al., submitted）, which is discussed in this paper. 6） On the basis of the findings indicating the close genetic relationship among rotaviruses of humans and various animal species, the possibility of interspecies transmission and subsequent genome reassortment in the recipient host and the significance of these events in the mechanisms of evolution and diversification of rotavirus in nature are discussed.

Group A rotaviruses (RVA) is the most important cause of severe gastroenteritis among children worldwide. Vaccination is considered the best alternative among public health measures to reduce and prevent the global burden caused by RVA infections. Rotarix™, a monovalent vaccine based on a human strain with a G1P[8]-1 specificity, was introduced in the National Brazilian Immunization Programs (NIP) in March, 2006. RVA P[8] is the most prevalent P genotype worldwide and four distinct phylogenetic lineages: P[8]-1, -2, -3, and -4 have been described. In the current study phylogenetic analysis of the VP8⁄ gene of 135 RVA P[8] Brazilian strains, in combination with G1, G3, G5 or G9 VP7 genotype, collected from 1986 to 2011 were carried out for a better understanding of the evolution of this viral genotype in Brazil. Lineages P[8]-1, P[8]-2, and P[8]-3 were observed circulating in Brazil. In 2001 these three P[8] lineages co-circulated simultaneously and this is the first report in South America to date. Considering the P[8] lineage and the G genotype, all G3 strains were related to lineage P[8]-3, whereas the G9 strains were related to P[8]-2 and P[8]-3 and G1 and G5 were related to P[8]-1, P[8]-2, and P[8]-3. In addition, the phylogenetic analysis based on estimate of genetic distances between P[8]-3 strains and the definition of a 1.5% cutoff value (with relevant statistical support) it was possible to propose a new classification for the P[8]-3 lineage into six different sub-lineages: P[8]-3.1 to P[8]-3.6. These findings reinforce the notion of the existence of constraints within specific RVA strains populations. The results obtained in this study reinforce the importance of a continuous RVA surveillance of circulating strains in order to predict the possible variants that will circulate in a country, assess the effects of vaccination on RVA circulating strains, and ultimately help in the design, challenges, and prospects of RVA vaccines.

A cross-sectional study was carried out in a dairy goat herd, investigating the presence of rotavirus by means of ELISA, polyacrylamide gel electrophoresis (PAGE) and two latex agglutination tests in feces of 63 goat kids younger than 1 month, with and without diarrhea, and in feces of 19 adult goats during the first few days after parturition. All animals belonged to a herd located in the mountains of the León province (NW Spain). Rotaviruses were found in 18 out of 63 goat kid fecal samples but no significant association between shedding of rotavirus and presence of diarrhea could be established. Rotaviruses were found in kids aged 6 to 21 days, and more frequently between 6 and 10 days. No shedding of virus was detected in any of the adults. Considering ELISA as the reference test, PAGE and both latex agglutination tests were less sensitive. One of the latex tests was also highly non-specific. All PAGE-positive samples showed the typical electropherotype of group A rotavirus. Feces were also screened for other pathogens includingSe realizó un estudio de corte en una explotación de ganado caprino lechero investigando la presencia de rotavirus mediante ELISA, electroforesis en gel de poliacrilamida (PAGE) y 2 pruebas de aglutinación en látex en las heces de 63 cabritos menores de un mes, con y sin diarrea, y en las heces de 19 cabras durante los primeros días tras el parto. Todos los animales pertenecían a una explotación situada en la montaña de la provincia de León (NO de España). Se encontraron rotavirus en 18 de las 63 muestras fecales de cabritos analizadas, aunque no pudo demostrarse la existencia de asociación estadísticamente significativa entre la eliminación del virus y la presencia de diarrea. Las edades de los cabritos en que se detectó eliminación de rotavirus estaban comprendidas entre los 6 y los 21 días y más frecuentemente entre los 6 y los 10. Considerando ELISA como la prueba de referencia tanto PAGE como ambas pruebas de aglutinación en látex fueron menos sensibles. Una de las pruebas de aglutinación en látex fue también muy inespecífica. En todas las muestras positivas mediante electroforesis se observó el electroferotipo característico de los rotavirus del grupo A. También se estudió la presencia en las heces de otros patógenos como

Rotavírus do grupo A (RVA) são reconhecidos como os principais agentes virais da gastroenterite aguda infantil e, devido aos relevantes índices de morbi-mortalidade, a vacinação tem sido considerada a melhor alternativa para a prevenção e o controle de RVA. No Brasil, em março de 2006 o Ministério da Saúde adicionou ao Programa Nacional de Imunização a Vacina Oral de Rotavírus Humano (VORH), a qual foi desenvolvida a partir de uma amostra monovalente atenuada G1P[8]. Neste contexto, o presente estudo objetivou a investigação da ocorrência das infecções por RVA na cidade de Goiânia após a implantação da VORH, bem como proceder a uma análise comparativa das amostras de RVA circulantes nos períodos pré e pós-vacinal. Para identificação de RVA, 65 espécimes fecais coletados no período de 2008 a 2009 de crianças com gastroenterite aguda, foram submetidos ao Ensaio Imunoenzimático e Eletroforese em Gel de Poliacrilamida, sendo observado um índice total de detecção de 16,9% (11/65). Após caracterização molecular, o genótipo G2 foi identificado em 10 amostras, sendo que quatro destas foram definidas como G2P[4]. Para análise comparativa, as amostras G2P[4], bem como outras 15 amostras coletadas nos períodos pré e pós-vacinal, foram submetidas ao sequenciamento genômico para as regiões codificantes das proteínas VP6, VP7 e NSP4. A caracterização molecular do gene de VP7 mostrou que as amostras G1 pertenciam às linhagens I e II, sublinhagens d e b, respectivamente, e que todas as amostras G2 pertencem à linhagem II, com a diferenciação de três sublinhagens, a, c e d, as quais foram correlacionadas com os períodos de coleta. Considerando os genogrupos de VP6 e genótipos de NSP4 identificados, observou-se predominância para genogrupo I e genótipo A no período pós-vacinal, enquanto, genogrupo II e genótipo B foram identificados com maior frequência antes da implantação da vacina. A associação entre os genótipos G e P com genogrupos de VP6 e genótipos de NSP4 revelou predominância da combinação G1-P[8]II-B no período pré-vacinal e da associação G2-P[4]-I-A no período pós-vacinal o que sugere uma substituição destas combinações após a implantação da VORH. The group A rotaviruses (RVA) are recognized as the main viral agents of acute childhood gastroenteritis, and due to the high morbidity-mortality rates vaccination has being considered the best alternative for prevention and control of RVA. In Brazil, in March, 2006 the Ministry of Health included the human rotavirus oral vaccine – VORH, which was developed from a monovalent attenuated strain G1P[8], in the National Immunization Program. In this context, the present study aimed at the investigation of the occurrence of RVA infections in the city of Goiânia after the implementation of the VORH, as well as the comparative analysis of the RVA circulating strains during the pre- and post-vaccination periods. For the RVA identification, 65 fecal samples obtained from children with acute gastroenteritis, in the period from 2008 to 2009, were tested by an immunoenzymatic assay and by polyacrilamide gel electrophoresis, with a total detection rate of 16.9% (11/65). After molecular characterization, the G2 genotype was identified in 10 samples, and four of those were considered as G2P[4] genotype. For the comparative analysis, the G2P[4] samples, as well as other 15 samples, obtained in the pre- and post-vaccination periods, were submitted to genomic sequencing of the coding regions for the proteins VP6, VP7 and NSP4. The molecular characterization of the VP7 gene showed that the G1 samples belonged to lineages I and II, sublineages d and b, respectively, and that all the G2 samples belonged to lineage II, with the differentiation of three sublineages, a, c and d, which were correlated with the collection periods. Regarding the VP6 genogroups and the NSP4 genotypes, a predominance of genogroup I and genotype A in postvaccination period was observed, whereas a predominance of genogroup II and genotype B was identified in the period before de vaccine implementation. The association between the G and P genotypes with VP6 genogroups and NSP4 genotypes revealed the predominance of the G1-P[8]-II-B combination in the pre-vaccination period, and the association G2-P[4]-I-A in the post-vaccination period, which suggests the substitution of these combinations after the implementation of the VORH. Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq

We report here the full genomic analysis of a simian SA11-like G3P[2] group A rotavirus (GAR) strain, B10, isolated from an asymptomatic infant in Kenya in 1987. By nucleotide sequence identities and phylogenetic analyses, the VP7–VP4–VP2–VP3–NSP1–NSP2–NSP3–NSP5 genes of strain B10 exhibited maximum genetic relatedness to those of the different isolates of simian strain SA11, and were assigned to the G3–P[2]–C5–M5–A5–N5–T5–H5 genotypes, respectively. On the other hand, the VP1, VP6 and NSP4 genes of strain B10 did not belong to any of the established GAR genotypes, and therefore, were assigned to new genotype numbers R8, I16 and E13, respectively, by the Rotavirus Classification Working Group. These observations suggested that strain B10 might have originated from reassortment event/s involving simian SA11-like strains and GAR strains from unknown animal host species (possibly other wild animals) preceding transmission to humans. Alternatively, considering the lack of data on simian GARs, it might be also possible that the VP1, VP6 and NSP4 genes of strain B10 are those of unknown simian strains, and that strain B10 might be a typical simian strain that was directly transmitted to humans. Therefore, either hypothesis pointed towards a rare instance of possible direct transmission of GARs from an animal host (possibly a monkey or some other wild animal) to humans. This was corroborated by the presence of different species of wild animals including non-human primates, and unhygienic conditions at the sampling site. To our knowledge, the present study is the first report on the detection of a simian SA11-like G3P[2] GAR strain in humans.

A total of 332 fecal samples were collected from 245 diarrheic as well as 75 recovered and 12 healthy camel calves in four different areas in Sudan [north (River Nile), east (Gedarif), central to south (Sennar, Blue Nile) and west (Kordofan) States]. Using ELISA, 46 samples (13.9%) were found positive for group A rotavirus. Out of 46 ELISA positive calves, we found 35 diarrheic up to 3 months of age, 7 recovered 4-6 months of age and 4 clinically healthy 8-12 months of age. Using EM, 6 out of 22 ELISA positive samples showed the well defined wheel-like structure characteristic of rotavirus. The results indicate the significant role of rotavirus in the epidemiology of camel calf diarrhea in Sudan. Attempts to isolate the camel group A rotavirus in MA104 cell culture were carried out on 21 ELISA-positive samples with successful isolation of the virus in 18. The isolated viruses were identified by ELISA and EM. Cytopathic effects (CPE) were manifested as rounding, elongation, triangulation, vacuolation and granulation of cells while the cell sheet remains intact. The CPE appeared on days 3-5 on the first to the second passages after treatment of the inocula and the cell culture media with trypsin. To our knowledge this is the first report on the isolation of camel rotavirus in cell culture., Prikupljena su 332 uzorka fecesa od 245 devine teladi s proljevom, 75 teladi oporavljene od proljeva i 12 od zdrave devine teladi sa četiri različita područja u Sudanu: sjevernoga (područje rijeke Nila), istočnoga (područje Gedarif), središnjega i južnoga (područje Sennar, Plavi Nil) te zapadnoga područja (Kordofan). Pretragom imunoenzimnim testom 46 uzoraka (13,9%) bilo je pozitivno za skupinu A rotavirusa. Od tih 46 pozitivnih životinja imunoenzimnim testom, 35 s proljevom bilo je u dobi do tri mjeseca, sedam s preboljelim proljevom bilo je u dobi od četiri do šest mjeseci, a četiri životinje u dobi od osam do 12 mjeseci bile su klinički zdrave. Elektronskom mikroskopijom dokazane su čestice karakteristične za rotavirus u šest od 22 uzorka pozitivna pretragom imunoenzimnim testom. Rezultati ukazuju na znatnu ulogu rotavirusa u epizootiologiji proljeva u devine mladunčadi u Sudanu. Skupina A rotavirusa izdvojena je na MA104 staničnoj kulturi iz 18 od 21 uzorka pozitivnoga pretragom imunoenzimnim testom. Izdvojeni virusi bili su identificirani imunoenzimnim testom i elekronskom mikroskopijom. Citopatski učinak očitovao se zaokruživanjem stanica, njihovim izduživanjem, triangulacijom, vakuolacijom i granulacijom, dok je stanični sloj ostao netaknut. Citopatski učinak javljao se za tri do pet dana u prvoj i drugoj pasaži nakon obradbe inokula i staničnog medija tripsinom. Ovo je prvo izvješće o izdvajanju rotavirusa deva u staničnoj kulturi.

Dual infections by group A and group C rotaviruses have been reported, but no reassortants between group A and group C rotaviruses have been described. The VP6 major inner capsid protein of group A and C rotaviruses shares common antigens detected by monoclonal antibodies and also shares 40–43% amino acid identity. Coinfection of Spodoptera frugiperda (Sf9) insect cells with different combinations of the recombinant baculoviruses encoding either group A {RF VP2 (A-VP2), IND VP6 (A-VP6), and VP7 (A-VP7[IND]), 2292B VP7 (A-VP7[2292B])} or C [Shintoku VP6 (C-VP6) and VP7 (C-VP7)] bovine rotavirus proteins produced hybrid group A/C triple-layered VP2/6/7 virus-like particles (TLPs) composed of A-VP2/C-VP6/C-VP7, A-VP2/C-VP6/A-VP7(IND), A-VP2/C-VP6/A-VP7(2292B), and A-VP2/A-VP6/C-VP7. To our knowledge, this is the first report that the inner capsid VP6 of group A or group C rotavirus can support attachment of the heterologous, antigenically distinct group A (G6, IND or G10, 2292B) or group C rotavirus outer capsid VP7 to produce hybrid TLPs in vitro.

Reverse transcription (RT)-PCR is now the standard method for typing group A rotaviruses (RVA) to monitor the circulating genotypes in a population. Selection of primers that can accurately type the circulating genotypes is crucial in the context of vaccine introduction and correctly interpreting the impact of vaccination on strain distribution. To our knowledge this study is the first report from Asia of misidentification of genotype G9 as G3 due to a primer-template mismatch. We tested two published G-genotype specific primers sets, designed by Gouvea and colleagues (Set A) and Iturriza‐Gomara and colleagues (Set B) on RVA from Hong Kong and Sri Lanka. Among 52 rotaviruses typed as G3 by set A primers, 36 (69.2%) were identified as G9 by nucleotide sequencing and set B primers. Moreover, of 300 rotaviruses tested, 28.3% were untypable by set A primers whereas only 12.3% were untypable by set B primers. Our findings reinforce the need to periodically monitor the primers used for RVA genotyping.