Detection and molecular characterisation of rotavirus and norovirus infections in Jordanian children with acute gastroenteritis

Rotavirus and norovirus are globally important causes of paediatric gastroenteritis, but no studies of viral genotypes have been reported from Jordan. We undertook a molecular epidemiological study in children hospitalised with acute gastroenteritis in Jordan between January 2006 and December 2007. Among 368 children, rotavirus and norovirus infections were detected in 49.5% and 11.4% of children, respectively. Rotavirus genotypes P[8],G1 (56%), P[4],G2 (14%) and P[8],G9 (13%) were most commonly identified, consistent with results of global rotavirus surveillance studies. Norovirus GII.3 was the most commonly detected genotype, followed by GII.4, contrasting with most studies in which GII.4 has predominated.     

Diversity of human rotavirus G9 among children in Turkey

Between September 2004 and December 2005 a prospective study was conducted to understand the epidemiology of rotavirus infection among children with diarrhea attending two hospitals in Ankara, Turkey. Rotavirus was detected in 39.7% of the 322 stool samples and affected mainly children in the age group of 6-23 months. More than 70% and 39% of these cases occurred in children <2 and <1 year of age, respectively. In the temperate climate of Ankara rotavirus infection was prevalent throughout the year. Serotype G1P[8] was dominant followed by G9P[8]. In 38 samples a total of 5 electropherotypes were detected. All G9P[8] were of long electropherotype except one of short electropherotype. A proportion of G1 and G9 strains were in combination with P[6], P[4] or P nontypable. Mixed serotypes were responsible for 2.4% of the infections. A phylogenetic tree constructed with the deduced amino acid sequences of the VP7 gene showed that 16 Turkish G9 strains clustered with rotaviruses of lineage III. One G9 strain formed a new lineage, lineage IV with the Sri Lankan G9 rotaviruses. In the phylogenetic tree of the VP8* gene, the Turkish G9P[6] rotaviruses clustered with human strains of lineage Ia. Increased diversity of the G/P type combination and the presence of infection throughout the year in Turkey was a situation similar to developing countries. The occurrence of rotavirus infection at later age and low level of mixed infections in Turkey represented the situation of developed countries. This study suggests that diverse G9 rotaviruses are emerging in Turkey.     

Molecular epidemiologic analysis of group A rotaviruses in adults and children with diarrhea in Wuhan city,China, 2000–2006

Summary To compare epidemiologic features and genetic characteristics of group A rotaviruses causing diarrhea in children and adults, a survey was conducted in Wuhan, China, during the period of Dec. 2000–May 2006. A total of 3839 stool specimens from diarrheal patients from eight hospitals were analyzed. Winter seasonality was observed for rotavirus diarrhea in both adults and children, showing overall rotavirus-positive rates of 9.0 and 23.9%, respectively. Throughout the study period, G3 was the most frequent G serotype in both adults and children (detection rates 86.2 and 87.8%, respectively), and was mostly associated with VP4 genotype P[8], VP 6 genotype II (subgroup II), and NSP4 genotype B. G3 rotaviruses were differentiated into eight electropherotypes, among which seven types were found in specimens from both adults and children. VP7 gene sequences of G3 rotaviruses from adults and children (6 and 4 strains, respectively), detected in different years and different hospitals, showed extremely high sequence identities (99–100%) to each other and to a few G3 rotavirus strains reported in Asia. However, lower sequence identities (82–96%) were observed to most of the human and animal G3 rotaviruses reported so far, including some Chinese strains. These findings indicate that in Wuhan, China, epidemic and genetic features of rotaviruses are similar in adults and children, and it has been suggested that G3 rotaviruses that might have originated from the same rotavirus were circulating among children and adults as prevailing viruses. In this study, two rotavirus strains, G9P[8] strain L169, derived from an adult, and G4P[6] strain R479, derived from a child, were isolated and genetically analyzed. The VP7 gene of L169 belongs to a major lineage of G9 rotaviruses that are globally widespread, but is distinct from G9 rotaviruses reported previously in China. The strain R479 had a VP7 gene which was divergent from most G4 human rotaviruses and showed an unusual dual subgroup specificity, I + II. The R479 VP6 gene does not belong to the main clusters of subgroup I and II rotaviruses phylogenetically, but is related to those of the porcine rotaviruses and some unusual human rotaviruses represented by the RMC321 strain isolated in eastern India.     

A hospital based study on inter- and intragenotypic diversity of human rotavirus A VP4 and VP7 gene segments,Germany

BackgroundEfforts to reduce the impact of group A rotaviruses on human morbidity and mortality rely on oral immunisation with live attenuated or recombinant vaccines. A major challenge in immunisation is the vast inter- and intragenotypic diversity accomplished by circulating rotaviruses.ObjectivesTo monitor rotavirus inter- and intragenotypic diversity in hospitalised children.Study designFrom January 2008 to December 2009 stool samples from 1994 paediatric in-patients suffering from diarrhoea were screened for rotavirus. Rotavirus G- and P-genotypes were determined by nucleotide sequencing and phylogenetic analysis was performed.ResultsRotavirus A was detected in stool samples of 341 children, comprising G1P[8], G2P[4], G3P[8], G4P[8], G9P[8], as well as uncommon G12P[6] genotypes and mixed infections. Predominant strains shifted from G1P[8] and G9P[8] genotypes in the first season to G3P[8] and G4P[8] genotypes in the second season. The highest intragenotypic diversity was detected in G1 strains and consisted of co-circulating G1-Ic, G1-Id, G1-Ie and G1-II rotaviruses. The G2 analysis revealed different intragenotypic lineages: G2-IIa, G2-IIb and G2-IIc. Interestingly, the circulating G4-Ib rotaviruses were characterised by insertions of 3 or 6 additional coding nucleotides within variable region 4 of VP7. Whereas different G9-III VP7 gene segments were detected G3-Ia sequences were highly homologous. In the VP4 analysis P[8]-III gene segment predominated over P[4]-Vb, P[8]-I, P[8]-IV and P[6]-I.ConclusionsA remarkable rotavirus heterogeneity was detected in the limited local setting and time span. Continued monitoring and nucleotide sequencing is necessary to document possible effects of rising immunisation levels on intragenotypic rotavirus diversity.     

Rotavirus infections in children in Turkey: A systematic review

We aimed to describe rotavirus epidemiology and clinical findings including extraintestinal manifestations in a setting that has yet to introduce rotavirus vaccines in the national immunization program. A literature search was performed by using the key words “Turkey” and “rotavirus.” Ninety‐eight studies published between 1987 and 2016 including epidemiological, clinical, and genotypical data at least 1 year duration were included. There were a total of 117 741 children with diarrhea and 26 566 rotavirus gastroenteritis with a median detection rate 31.8% (95% CI, 31.3‐32.4) under 5 years of age. The rate of dehydration was 47% (95% CI, 23.4‐91.6). There were 328 cases reported to be presenting with a various complication related to rotavirus in 2750 children in eight studies. The overall complication rate was 11.7% (95% CI, 10.7‐12.9). The cumulative incidence of the most common genotypical combinations circulating worldwide was only 59.7% (G9[P8], 25%; G1[P8], 22%; G2[P4], 5.6%; G3[P8], 2.6%; G4[P8], 4.5%) whereas mixed, untypeable, and other genotypes were 2.4%, 15%, and 22.9% respectively. Our results point out the importance of rotavirus vaccination by presenting that rotavirus may cause severe complications besides severe gastroenteritis. The role of strain diversity in the variability of clinical presentations of rotavirus infections needs to be further investigated.     

Non-structural protein NSP2 induces heterotypic antibody responses during primary rotavirus infection and reinfection in children

Rotaviruses are the single most important causes of severe acute diarrhoea in children worldwide. Despite success in developing vaccines, there is still a lack of knowledge about many components of the immune response, particularly those to non-structural proteins. This study established radioimmunoprecipitation (RIP) assays using labeled G1P[8], G2P[4], and G4P[6] human rotaviruses to examine the spectrum and duration of rotavirus antibodies in sera collected sequentially for 18-36 months from 27 children after hospitalization for primary rotavirus gastroenteritis. Five children experienced rotavirus re-infections. Primary responses detected to non-structural protein NSP2 declined to baseline after 100-150 days. Responses were heterotypic between NSP2 of G1P[8] and G4P[8] rotaviruses. Re-infections after 465-786 days boosted antibody levels to NSP2of both serotypes, together with the appearance of anti-NSP2 to G2P[4], even though there was no evidence of infection with this serotype. We developed an enzyme-immunoassay to measure sequential levels of anti-NSP2 IgG and IgA, using recombinant (heterotypic) NSP2 derived from SA11 (G3P[2]). Anti-NSP2 IgG and IgA were detected in sera from 23/23 (100%) and 18/24 (75%) of children after primary infection, declined to baseline after 100-150 days, were boosted after rotavirus re-infections, and again declined to baseline 150 days later. Anti-NSP2 IgA was also detected after primary infection, in duodenal juice from 14/16 (87%), and faecal extract from 11/19 (57%) of children. Sequential estimation of anti-NSP2 EIA levels in sera could be a sensitive index of rotavirus infection and re-infection. The potential of anti-NSP2 to limit viral replication after re-infection deserves further study.     

Rotavirus genotype distribution in Kyrgyzstan and Kazakhstan, 2007–2009

This is the first study to present rotavirus genotype distribution in children admitted to a hospital with acute gastroenteritis in Kyrgyzstan and Kazakhstan from January 2007 through December 2009. In total, 858 rotavirus ELISA‐positive samples were characterized by RT‐PCR, with a considerable geographical and seasonal variation in genotype distribution observed during the study. The globally common genotypes (G1P[8], G2P[4], G3P[8], G4P[8], G9P[8], G12P[8] and G12P[6]) accounted for 81.5–88.2% of the infections in Kyrgyzstan and 72.3–79.3% of the infections in Kazakhstan. The predominant genotypes were G1P[8], G2P[4] and G3P[8]. G1P[8] was the dominating genotype in Kyrgyzstan, detected in 51–64.7% of the samples. A similar predominance was not seen for G1P[8] in Kazakhstan, with a shift to G2P[4] predominance being seen in 2008. G9P[8] was a rare genotype in both countries, whereas G12 was detected in between 2.2% and 7.6% of the samples. The surveillance period was characterized by many co‐circulating genotypes, and eight unusual combinations (G1P[4], G2P[8], G2P[6], G3P[4], G9P[4], G12P[4], G9P[9] and G10P[4]) were detected. This study provides important baseline data on rotavirus genotypes in Kyrgyzstan and Kazakhstan in the pre‐vaccine era, and the results may indicate that the two licensed vaccines can be expected to prevent rotavirus disease in these countries.     

Predominance of genotype P[9]G3 in rotavirus gastroenteritis in Polish children

Introduction

Rotavirus (RV) infection is the most common cause of gastroenteritis in children. This paper identifies the most common genotypes of rotaviruses isolated from children hospitalized with gastroenteritis and attempts to determine any relationship between infection with a certain rotavirus genotype.

Material and methods

The investigated group consisted of 68 consecutive children with rotavirus gastroenteritis (confirmed by an agglutination test). Rotavirus genotype was determined in stool samples obtained from each child.

Results

The P[9]VP4 genotype was observed in 41/61 positive samples (over 67.2%) that were permanently associated with the G3 VP7 genotype. Moreover, G3 was determined as the most commonly isolated G type (77.94%). As well as the P[9]G3 type, G3 was also found in the P[4] type (5 cases). Twenty-six out of 61 (42.6%) children in whom rotavirus genotype was determined were co-infected with pathogenic bacteria. No statistical correlation was observed between rotavirus P[9]G3 gastroenteritis and digestive tract co-infection with pathogenic bacteria (p > 0.05). Elevated ALT activity was found in 34/59 (57.6%) cases of rotavirus gastroenteritis. Elevated ALT serum level was found to correlate with P[9]G3 rotavirus genotype but concomitant infections did not.

Conclusions

The most common genotype of rotaviruses observed in our group of children, P[9]G3, has rarely been described. Co-infection of the digestive tract with pathogenic bacteria and elevated serum ALT concentrations were found to be the most frequent phenomena. A correlation between P[9]G3 rotavirus genotype and elevated serum ALT level was found, but no significant relationship was identified between concomitant infections and P[9]G3 genotype.     

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. G1P[8] strains were predominant (44.0%), followed by G9P[8] (17.7%), G3P[8] 13.1%, G4P[8] (9.5%), and G2P[4] (1.8%); mixed rotavirus infections occurred in 2.3%. G9 rotaviruses emerged during the 2004–2005 season (73.4%) and remained the second most prevalent strains. Few unusual strains, G6, G8, G12 and P[6]-types, were detected. The monitoring of rotavirus infections should be maintained to document strain distribution and to assess the emergence of new reassortants that may not respond to current rotavirus vaccines.     

Phylogenetic analysis of rotaviruses with genotypes G1, G2, G9 and G12 in Bangladesh: evidence for a close relationship between rotaviruses from children and adults

To clarify the phylogenetic relatedness of rotaviruses causing gastroenteritis in children and adults, an epidemiologic investigation was conducted in Mymensingh, Bangladesh, during the period between July 2004 and June 2006. A total of 2,540 stool specimens from diarrheal patients from three hospitals were analyzed. Overall, rotavirus-positive rates in children and adults were 26.4 and 10.1%, respectively. Among the 155 rotavirus specimens examined genetically from both children and adults, the most frequent G genotype was G2 (detection rate: 54.0 and 47.6%, respectively), followed by G1 (21.2 and 26.2%, respectively), and G9 (15.9 and 9.5%, respectively). G12 was also detected in five specimens (3.2% in total; four children and one adult). Sequence identities of VP7 genes of G2 rotaviruses from children and adults were higher than 97.8%, while these Bangladeshi G2 viruses showed generally lower identities to G2 rotaviruses reported elsewhere in the world, except for some strains reported in African countries. Similarly, extremely high sequence identities between children and adults were observed for VP7 genes of G1, G9 and G12 rotaviruses, and also for the VP4 genes of P[4], P[6], and P[8] viruses. Rotaviruses from children and adults detected in this study were included in a single cluster in phylogenetic dendrograms of VP7 or VP4 genes of individual G/P types. Rotaviruses with two emerging types, G9 and G12, had VP7 genes that were phylogenetically close to those of individual G-types recently reported in Bangladesh and India and were included in the globally spreading lineages of these G-types. These findings suggested that genetically identical rotaviruses, including those with the emerging types G9 and G12, were circulating among children and adults in city and rural areas of Bangladesh.     

Evaluation of a multiplex real time reverse transcription PCR assay for the detection and quantitation of the most common human rotavirus genotypes

Group A rotaviruses (RVs) are important pathogens that cause acute, dehydrating gastroenteritis in infants and young children. In this study, a multiplex real-time polymerase chain reaction protocol using primers and TaqMan(?) probes specific for viral VP4 and VP7 genes was evaluated. This assay offers simultaneous genotyping and quantification of the most common RV genotypes G1P[8], G2P[4], G3P[8], G4P[8], and G9P[8]. It was compared to the molecular typing results provided by conventional PCR. A total of 92 archived stool specimens obtained from children younger than 5 years old with the diagnosis of acute gastroenteritis were examined. Real-time PCR assay detected rotavirus strains among the most common genotype combinations G4P[8] (70.7%), G1P[8] (10.9%), G2P[4] (5.4%), G9P[8] (2.2%). This new assay described has an acceptable sensitivity (low limit 6.3×10(2)copies/g of stool).     

Changing pattern of rotavirus G genotype distribution in Chiang Mai, Thailand from 2002 to 2004: decline of G9 and reemergence of G1 and G2

Group A rotaviruses are the most common cause of acute viral diarrhea in humans and animals throughout the world. Previous surveillance studies of group A rotaviruses in Thailand indicated that the dominant types of rotaviruses were changing from time to time. During 2000 and 2001, the G9 rotavirus emerged as the most prevalent genotype, with an exceptionally high frequency (91.6%) in Chiang Mai, Thailand. In the year 2002-2004, group A rotavirus was detected in 98 out of 263 (37.3%) fecal specimens collected from children hospitalized with diarrhea. Of these, 40 (40.8%) were G9P[8], 33 (33.7%) were G1P[8], 23 (23.5%) were G2P[4], and 2 (2.0%) were G3P[9]. The G9P[8] was found to be the most predominant strain in 2002, but the prevalence rate abruptly decreased during the period 2003-2004. In addition, G2P[4] reemerged in the epidemic season of 2003, whereas G1P[8] became the most predominant strain in the following year (2004). Phylogenetic analysis of the VP7 genes revealed that G1, G2, and G9 rotavirus strains clustered together with recently circulating strains, which were isolated from different regional settings in Thailand. In conclusion, the study demonstrated a decrease of incidence of G9P[8] and reemergence of G1P[8] and G2P[4] rotaviruses in Chiang Mai, Thailand during the period 2002-2004. These data imply that the distribution of group A rotavirus genotypes circulating in Chiang Mai, Thailand, changes over time.     

Epidemiological features of rotavirus infection among hospitalized children with gastroenteristis in Ho Chi Minh City,Vietnam

An epidemiological study of the G serotype and P genotype distribution of group A rotaviruses by using ELISA and/or RT-PCR was conducted in children (aged 1 month to 15 years) with diarrhea that were admitted to the General Children's Hospital No. 1, Ho Chi Minh City, Vietnam from December 1999 to November 2000. The results showed that rotavirus is associated with 65.6% (889/1355) of diarrheal admissions. Rotavirus infection mostly affected children under 2 years of age with a peak incidence in children 1 to 2 years of age (75.7%) and it occurs year round with a slight seasonal pattern; 99.5% of the specimens could be G-typed: G1 was predominant (68.7%), followed by G4 (15.4%), G2 (12.3%), G3 (0.6%), and G9 (0.5%). High identities of VP7 nucleotide (96.3 to 96.9%) and deduced amino acid (98.1 to 98.4%) were found between two Vietnamese G9 strains and also the recent emergence of G9 strains US 1205, Brazilian R143, and Malawian MW69. Mixed infections were identified in 17 (2.0%), and 5 strains (0.5%) remained untypable. The four most common worldwide strains, G1P[8], G2P[4], G3P[8], and G4P[8], constituted 81.1% of all rotaviruses typed with G1P[8] being the most prevalent type (58.2%). Unusual G/P combinations (11 strains) were detected in 11.7% of all strains, of which, G1P[4] was the most prevalent, accounting for 5.6% of the total. Several combinations of G and P types were observed in this study, suggesting a complex rotavirus infection pattern in Vietnam. This study has provided for the first time clear indication on the circulating G and P genotypes among hospitalized children in Ho Chi Minh City, Vietnam. The results suggest that these viral infections are prevalent among hospitalized children and that the four most common worldwide G types as well as the four most common G-P combinations were also infecting children in Ho Chi Minh City, Vietnam. This result could have important implications for rotavirus vaccine programs and for understanding the epidemiological characteristics of human rotavirus in Ho Chi Minh City, Vietnam.     

Molecular epidemiology of group A rotavirus in outpatient diarrhea infants and children in Chongqing,China, 2011-2015

Human group A rotavirus (RVA) is the leading cause of acute viral gastroenteritis in children under 5 years old worldwide. The aim of this study was to investigate the genotype distribution of RVA in the Midwest of China. Sentinel-based surveillance of acute diarrhea was conducted at Children's Hospital of Chongqing Medical University from 2011 to 2015. RVA was tested by using enzyme-linked immunosorbent assays. The partial VP4 genes and VP7 genes of rotavirus were amplified and sequenced, and genotyping and phylogenetic analyses were performed. Among the 2236 stool specimens collected from children with acute gastroenteritis, 681 (30.46%) were positive for RVA. The majority of children (89.28%) who tested positive for RVA were children aged ≤2 years. The seasonal peak of RVA was in the winter. As for genotype, four strain combinations, G9P[8], G3P[8], G1P[8], and G2P[4] contributed to 75.62% (515/681) of the RVA-associated diarrhea cases. After a marked increase in G9P[8] (30.77%) in 2013, G9P[8] became the predominant genotype in 2014 and 2015, whilst the prevalence of G1P[8] was decreased to 2.72% in 2015. Unusual G-P combinations (eg, G1P[4], G9P[4], G4P[6], G3P[4], G2P[8]) were also detected sporadically over the study period. Phylogenetic tree analysis results showed that the VP7 sequences of G9 strains were clustered into two main lineages, and 77.34% of them were clustered into lineage VI, with the highest nucleotide similarity to the strain JS12-17(China). VP4 gene sequences of P[8] strains were almost P[8]-lineage 3. Substantial temporal variation in the circulation of various genotypes of rotavirus in Chongqing was observed during 2011-2015, and highlights the need for continuous surveillance of RVA infection for better understanding and control of RVA infection.