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Epidemiological study of the G serotype distribution of group A rotaviruses in Kenya from 1991 to 1994.

An epidemiological study on the G serotype distribution of group A rotaviruses (GARV) isolated in Kenya was carried out in one urban hospital in Nairobi and in two rural hospitals in Nanyuki and Kitui to clarify the prevalent G serotypes before future introduction of the ready licensed rotavirus vaccine in Kenya. A total of 1,431 stool specimens were collected from children, who were mainly outpatients, aged from 0 to 6 years old with acute gastroenteritis from August 1991 to July 1994. Samples positive for GARV by conventional ELISA were then analyzed by subgrouping and serotyping ELISA and by PAGE. To ascertain the G serotypes of viruses in samples that were unable to be typed by serotyping ELISA, polymerase chain reaction was also attempted. The prevalence of GARV was 28.4% in the urban hospital, 22.5% in Nanyuki, and 13.7% in Kitui. Among rotavirus-positive samples, subgroup II rotaviruses were detected in 63.1%, and subgroup I rotaviruses were 25.9%. Serotype G4 was most prevalent, accounting for 41.6% followed by 23.3% of serotype G1, 17.0% of serotype G2, and serotype G3 was rarely isolated. Seven strains of serotype G8/P1B rotavirus was detected for the first time in Kenya by RT-PCR. Eleven specimens with an unusual composition of subgroup, serotype, and electropherotype were atypical GARV in which the P-serotype was P1A, P1B, or P2. Although uncommon GARV serotype G8/P1B and atypical GARV were detected, the four major GARV serotypes, G1 through G4, should be targeted at this moment for vaccination to control this diarrheal disease in Kenya. Continuous monitoring of the G- and P-serotype distribution of GARV should provide important information about the impact of rotavirus vaccination in Kenya.

Child↗

Pathogenesis of rotavirus gastroenteritis.

The outcome of intestinal infection with rotaviruses is more complex than initially appreciated, and it is affected by a complex interplay of host and viral factors. Rotaviruses infect intestinal enterocytes, and the early events in infection are mediated by virus-epithelial cell interactions. Diarrhoea may be caused by several mechanisms including (i) malabsorption that occurs secondary to the destruction of enterocytes, (ii) villus ischaemia and activation of the enteric nervous system that may be evoked by release of a vasoactive agent from infected epithelial cells in the absence of significant pathologic lesions or enterocyte damage, and (iii) intestinal secretion stimulated by the intracellular or extracellular action of the rotavirus non-structural protein, NSP4, a novel enterotoxin and secretory agonist with pleiotropic properties. New studies of rotavirus infection of polarized intestinal epithelial cells show that rotaviruses infect cells differently depending on whether or not they require sialic acid for initial binding, and infection alters epithelial cell functions. NSP4 also affects epithelial cell function and interactions. NSP4 (i) induces an age- and dose-dependent diarrhoeal response in young rodents that is similar to virus-induced disease, (ii) stimulates a Ca(2+)-dependent cell permeability where the secretory response is age-dependent, and (iii) alters epithelial cell integrity. Antibody to NSP4 protects mouse pups from diarrhoea induced by homotypic and heterotypic viruses. These data support a new mechanism of rotavirus-induced diarrhoea whereby a viral enterotoxin triggers a signal transduction pathway that alters epithelial cell permeability and chloride secretion. This new information about how a gastrointestinal virus causes disease demonstrates common pathogenic mechanisms for viral and bacterial pathogens not previously appreciated. These results also suggest new approaches to prevent or treat rotavirus-induced diarrhoea.

Animals↗

Neonatal rotavirus infection in Belém, northern Brazil: nosocomial transmission of a P[6] G2 strain.

A total of 614 fecal specimens were obtained during a survey for rotavirus infection conducted between May 1996 and May 1998 among 437 newborns admitted to special care nurseries at a public hospital in the urban area of Belém, Brazil. Routine stool samples were taken weekly from all babies up to the age of 28 days. Overall, 51 (11.7%) of the neonates excreted rotaviruses while in hospital, of whom 42 (82.3%) developed asymptomatic nosocomial infection; nosocomial infection was also proved in five of the nine patients with diarrhea. Three distinct RNA profiles were detected, of which one short electropherotyping pattern was far more frequent ( approximately 90% of the strains). Using monoclonal antibody-based enzyme immunoassays, 32 (62.7%) of the rotavirus-positive strains were classified as G2, and 1 (1.9%) as mixed G1 and G2. A G serotype could not be assigned to 18 (35.3%) of the isolates. A reverse transcription-polymerase chain reaction was used for determining the VP4 type-specificity of a subset of 28 rotavirus-positive samples. Characterization of the VP7-genotype specificity was also sought for 18 of these latter strains. Overall, P[6] and G2 genotypes were identified in 93% and 94% of tested samples respectively, with results being further confirmed by Southern hybridization. Although surveillance was conducted during a 25-month period, 50 (98%) of 51 rotavirus isolates clustered between January and December 1997. The earliest [P6]G2 rotavirus infections were detected by late January 1997, involving two (13- and 14-day-old) babies admitted with acute diarrhea. Thereafter, strains bearing these genotype specificities were identified among five infants with hospital-acquired gastroenteritis, followed by 16 others who were infected asymptomatically. This is the first report from Brazil describing nosocomial transmission of P[6]G2 rotavirus strains among neonates.

Blotting, Southern↗

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.

Adolescent↗

Genetic analysis of group B human rotaviruses detected in Bangladesh in 2000 and 2001.

Group B rotaviruses detected in Bangladesh in 2000 and 2001 were analyzed genetically to clarify relatedness to human group B rotaviruses reported previously in China and India, and to animal group B rotaviruses. VP7 gene sequences of the Bangladeshi group B rotaviruses (Bang373, Bang544, Bang334, and Bang402) were almost identical to each other and also showed high sequence identity to the Indian strain CAL-1 (98%) and Chinese strain adult diarrhea rotavirus (ADRV) (92%), while identities to bovine and murine viruses were considerably low (60-63%). Other genes of Bang373 and Bang544 encoding VP2, VP4, VP6, and NSP1 through NSP5 also showed much higher sequence identities to those of CAL-1 (97.7-99.4%) than to those of ADRV (89.9-93.9%). Characterization of nucleotide substitutions among Bang373, CAL-1, and ADRV suggested that all the gene segments might have evolved neutrally at similar mutation rates, while some of the gene segments (e.g., VP2 gene) were suggested to be more conserved than others. In conclusion, group B rotaviruses detected in Bangladesh represented by Bang373 and the Indian virus CAL-1 were considered as virtually identical viruses which are distinct genetically from ADRV, and it was suggested that Bang373 (CAL-1)-like group B rotavirus (Bengali strains) might be distributed primarily in an area around the Bay of Bengal.

Adult↗

Sequence analysis of genes encoding structural and nonstructural proteins of a human group B rotavirus detected in Calcutta, India.

Nucleotide sequences of RNA segments encoding structural proteins(VP4, VP6, and VP7) and nonstructural proteins(NSP1 and NSP3) of a human group B rotavirus CAL-1, which was detected in Calcutta, India, were determined and their relatedness with cognate genes of other group B rotaviruses was analyzed. The CAL-1 genes showed generally high sequence identities (more than 90%) to those of human group B rotavirus, adult diarrheal rotavirus (ADRV) in China, while identities with bovine, murine, and ovine viruses were considerably lower (58-73%). Among RNA segments analyzed, sequence identity of the VP6 gene was relatively high compared with other gene segments. In the CAL-1 VP7 sequence, many characteristics were shared by ADRV, but not by other animal group B rotaviruses. In contrast, VP4 and NSP3 of CAL-1 were single amino acid and 23 amino acids longer than those of ADRV strain, respectively, due to differences of a few nucleotides. These findings suggested that human group B rotaviruses CAL-1 and ADRV might have originated from a common ancestral virus distinct from animal group B rotaviruses reported so far, while some notable sequence differences indicated the distinct nature of these viruses.

3' Untranslated Regions↗

An outbreak of epidemic diarrhoea in adults caused by a new rotavirus in Anhui Province of China in the summer of 1983.

Shexian County of Anhui Province in East China experienced a severe outbreak of nonbacterial diarrhoea in May and June of 1983. Over 20,0000 cases were reported. Most were adults of 20 to 50 years old. The isolated virus was morphologically indistinguishable from ordinary infantile rotavirus, but CF and ELISA tests showed that the new virus lacked the common group-A antigen shared by ordinary rotavirus. Nine of 10 convalescent-phase and three paired sera of patients showed antibody positive, with a greater than four-fold antibody rise against new rotavirus in the CF test. Seventeen faecal samples from patients showed identical, and special, electropherotype. They all had a double-stranded RNA with 11 discrete segments. The RNA profiles were quite different from those of reference rotavirus Wa strain and ordinary infantile rotaviruses. Third and 4th segments were near to each other, 5th and 6th segments were very close, but 7th, 8th, and 9th segments were widely separated. This study indicates the new virus can be identified as part of a new group of rotaviruses. This new rotavirus has been incriminated as the causative agent of the epidemic.

Adult↗

Development of a lymphocyte transformation assay for rotavirus in whole blood and breast milk.

A multivariate assay was developed for detecting lymphoproliferative responses to rotavirus in whole blood and breast milk. Detection of lymphocyte transformation in samples from healthy uninfected adults required an assay incorporating a large number of antigen and cell concentrations and several incubation periods before pulsing. In contrast, lymphoproliferation to rotavirus was observed over a wider range of these variables in blood collected from a kidney recipient nine days post-rotavirus infection. Lymphoproliferation to rotavirus was detected in 32.5% of breast milk samples tested. The magnitude of lymphoproliferation to rotavirus did not correlate with rotavirus-specific IgG ELISA antibody titres in sera or with rotavirus-specific IgA ELISA antibody titres in breast milk.

Adult↗

Viraemia is a common finding in immunocompetent children with rotavirus infection.

Rotavirus infection is usually localized to the intestine but involvement of extra-intestinal sites, including the respiratory tract, liver, kidney, lymph nodes, and central nervous system, has been reported. The extra-intestinal spread of the virus may occur through blood since viraemia has been documented occasionally in animals and humans. Nevertheless, the questions of how common viraemia is in immunocompetent children and whether it is associated with extra-intestinal manifestations remain unsolved. Serum samples from 54 immunocompetent children hospitalized for acute diarrhea were evaluated prospectively for the presence of rotavirus RNA by nested reverse transcriptase-polymerase chain reaction (RT-PCR) to investigate this issue. Rotavirus antigens were detected in the stools of 14 children. A bacterial aetiology was documented in 14 cases, and in the remaining cases the aetiology remained unknown. Rotavirus RNA was detected in the blood of 9/14 (64.3%) rotavirus infected children but in no child with diarrhea of other origin. Positive RT-PCR was associated with high fever and/or evidence of extra-intestinal involvement. All positive samples were collected within 3 days of illness onset, suggesting that viraemia was detectable for only a few days. Children in whom rotavirus was detected only in stool samples had high fever but no other extra-intestinal feature. These data suggest that viraemia is common in children infected with rotavirus, which may be associated with extra-intestinal involvement.

Antigens, Viral↗

Rotavirus surveillance in the city of Rio de Janeiro-Brazil during 2000-2004: detection of unusual strains with G8P[4] or G10P[9] specificities.

Rotavirus diarrhea is a potentially life-threatening disease that affects millions of children annually around the world. Because protection against rotavirus disease is thought to be type specific, continuous rotavirus surveillance before and after implementation of a vaccine is still of essential importance. Rotavirus surveillance has been conducted in the city of Rio de Janeiro, Brazil since 1982. In the present study, we report rotavirus surveillance data in Rio de Janeiro city from 2000 to 2004. One hundred twenty nine of 1,568 (8.2%) stool samples, collected from children with acute diarrhea between January 2000 and July 2004 were rotavirus-positive. One hundred twenty eight of the 129 (99.2%) rotavirus-positive samples were genotyped for G and/or P specificity. G1 was the predominant strain (49.6%, 64/129) followed by G9 (30.2%, 39/129), and G4 (17.8%, 23/129); G2 and G3 viruses were not detected. One sample (0.8%) was non-typeable. P genotypes were determined for 124 of the 129 (96%) samples, and P[8] was the predominant genotype (90.6%, 117/129). Genotypes P[4] and P[9] were detected in two (1.6%) samples each; one (0.8%) sample presented P[6] genotype; and five (3.8%) samples were non-typeable. Two samples (1.6%) presented mixed P genotypes (P[6] + P[8]). Two unusual strains were isolated: a G8P[4] strain isolated from a non-hospitalized child with diarrhea and a G10P[9] strain isolated from a hospitalized child with diarrhea.

Brazil↗

A comparison of rotavirus strains of bovine, simian and porcine origin.

Three rotavirus strains of bovine, simian and porcine origin, respectively, were compared. The 3 viruses induced a classic rotaviral infection in newborn, conventionally reared calves. The cross neutralization tests revealed an antigenic identity of simian and porcine rotaviruses and a slight serologic correlation of these two viruses with the bovine rotavirus strain. However, in reciprocal cross protection tests carried out in calves, the simian rotavirus antiserum afforded weak protection to challenge infection with either the porcine or the bovine viruses. By contrast, the protective level of the bovine and the porcine rotavirus antisera was relatively high. It was speculated that the 81/36F bovine rotavirus could be considered, tentatively, as an antigenic reassortant rotavirus strain.

Animals↗

A family outbreak of gastroenteritis caused by group C rotavirus.

A family outbreak of gastroenteritis caused by group C rotavirus is described. All five members of the family, with children between 8 and 15 years of age, fell ill with diarrhea. The diagnosis was initially based on the detection of rotavirus RNA showing a typical group C profile in gel electrophoresis in stool samples, and it was serologically verified from patient sera using a cell culture adapted porcine group C rotavirus as a source of standard antigen. All collected serum samples from the family contained IgM and/or IgG class antibodies to group C rotavirus measurable by immunofluorescence antibody test (IFAT). Group C rotavirus specific IgM class antibodies were present in the early serum samples in 3/4 patients. A roller tube neutralization test (NT) was established to demonstrate neutralizing antibodies to porcine group C rotavirus in human sera. These methods can be used to detect serologically group C rotavirus infections.

Adolescent↗

Rotavirus gastroenteritis.

Rotavirus is the single most important cause of severe, dehydrating gastroenteritis in infants and young children worldwide. Rotavirus gastroenteritis is associated with high morbidity in developed countries and significant mortality in developing countries. Virtually all children are infected with rotavirus by 3 years of age. Fecal-oral transmission is the most likely route of virus spread. Group A serotype strains G1 through G4 account for more than 90% of rotavirus gastroenteritis in humans, with G1 being the predominant serotype. The virus preferentially infects the mature small-intestinal enterocytes. Rotavirus gastroenteritis is characterized by fever, vomiting, and diarrhea, with vomiting particularly prominent. Dehydration is a frequent complication because of the severity of the diarrhea and the associated vomiting. Rehydration and maintenance of proper fluid and electrolyte balance remain the mainstay of treatment. Hygienic measures have little effect on the reduction of rotavirus infection rates. The disease can be effectively controlled by universal rotavirus vaccination.

Child, Preschool↗

Antirotaviral activity of milk proteins: lactoferrin prevents rotavirus infection in the enterocyte-like cell line HT-29.

Different milk proteins were analyzed for their inhibitory effect on either rotavirus-mediated agglutination of human erythrocytes or rotavirus infection of the human enterocyte-like cell line HT-29. Proteins investigated were alpha-lactalbumin, beta-lactoglobulin, apo-lactoferrin, and Fe(3+)-lactoferrin, and their antiviral action was compared with the activity of mucin, a milk glycoprotein known to affect rotavirus infection. Results obtained demonstrated that beta-lactoglobulin, apo- and Fe(3+)-lactoferrin are able to inhibit the replication of rotavirus in a dose-dependent manner, apo-lactoferrin being the most active. It was shown that apo-lactoferrin hinders virus attachment to cell receptors since it is able to bind the viral particles and to prevent both rotavirus haemagglutination and viral binding to susceptible cells. Moreover, this protein markedly inhibited rotavirus antigen synthesis and yield in HT-29 cells when added during the viral adsorption step or when it was present in the first hours of infection, suggesting that this protein interferes with the early phases of rotavirus infection.

Animals↗

Genomic RNA constellation of recently emerging serotype G14 equine rotavirus strains in Japan that is highly homologous with prototype G3 and G14 strains previously identified in the United States of America.

Serotype G14 was once considered to be uncommon among equine rotaviruses. While it sporadically emerged in some parts of the world, serotype G14 became the dominant G serotype among rotaviruses detected in foals with diarrhea in Japan in the late 1990s. However, it is not known how such recently emerging G14 rotaviruses are related in their overall genomic RNA constellation to prototype G14 strain identified earlier in the United States of America or how they were generated and why they have dominated over G3 equine rotaviruses. Genogrouping by RNA-RNA hybridization revealed that recently emerging serotype G14 equine rotavirus strains had an overall genomic RNA constellation that was highly conserved not only with contemporary and earlier G3 strains in Japan but also with prototype G3 and G14 strains previously identified in the United States of America. Japanese G14 rotavirus strains are likely to have originated form a VP7 gene substitution reassortant that had been formed earlier in the United States of America on the background of the then dominant G3 equine rotavirus.

Animals↗

A lack of significant association between the electropherotype or G-serotype of the infecting strain and disease severity of rotavirus gastroenteritis.

Despite many previous studies, the question has not been settled as to whether some human rotavirus strains are more virulent than others. Since disease severity is most clearly reflected by the hospitalization status of the infected children, we examined whether there was any difference in the distribution of dominant strains between inpatient and outpatient groups. The study population comprised 763 children with acute diarrhea who were treated at a general hospital in Honjo City, Akita, Japan, during 1986-1997. Rotaviruses from stool specimens were classified into 77 electropherotypes using polyacrylamide gel electrophoresis. A single dominant strain or two co-dominant strains circulated simultaneously with some infrequent strains in most rotavirus seasons. Over the 11 rotavirus seasons, there was no significant difference in the relative frequencies of 15 rotavirus strains between the inpatient and the outpatient groups when strains of rotavirus were defined by their electropherotypes in polyacrylamide gel electrophoresis. However, infection with one G1 strain that co-dominated with a G4 strain carrying an identical electropherotype except the VP7 gene resulted in a statistically significantly reduced risk of hospitalization. There was no significant difference in the relative frequencies of four major G-serotypes or long/short RNA pattern. We conclude that the virulence or disease-causing potential of human rotavirus is not substantially different in the majority of strains.

Adolescent↗

Neutralization of rotavirus and recognition of immunologically important epitopes on VP4 and VP7 by human IgA.

Rotavirus is an important cause of gastroenteritis in young children. Locally produced antibodies in the intestinal mucosa are proposed to play an important role in the defence against rotavirus infection, but it is not established whether IgA alone can neutralize rotavirus, nor if IgA antibodies recognize epitopes involved in protective immunity. To evaluate whether human IgA plays a role in virus neutralization, serum IgA was purified from nineteen rotavirus seropositive individuals and examined for its neutralizing capacity by a peroxidase focus reduction test. In all nineteen sera IgA neutralizing antibodies against serotype 3 (rhesus rotavirus) were demonstrated. Purified IgA was further investigated and shown not only to neutralize rotavirus in solution but also to neutralize rotavirus already pre-bound to epithelial cells (MA-104). IgA epitope blocking assays with monoclonal antibodies directed against heterotypic epitopes on VP4 and VP7, revealed that IgA antibodies from 4/16 sera recognized epitopes on VP4, while 5/16 sera recognized a VP7 epitope. When whole sera were investigated for comparison 7 and 9/16 sera recognized epitopes on VP4 and VP7 respectively.

Adult↗

Determination of the duration of a primary immune response and the ID50 of ALA rabbit rotavirus in rabbits.

The rabbit model of rotavirus infection has been used to examine the immune response to rotavirus infection and to evaluate strategies for rotavirus vaccine development. To determine the 50% infectious does (ID50) of tissue culture adapted ALA virus, rabbit were orally inoculated with 10(1)-10(3) PFU of ALA rotavirus. The ID50 of ALA virus was determined to be 1.7 x 10(2) PFU. The immune response induced in rabbits infected at low virus doses (10(2)-10(3) PFU) was of similar magnitude to the immune responses induced with a high dose (10(6) PFU) inoculum, indicating that the immune response to ALA rotavirus in rabbits is not dose dependent. To determine if a single rotavirus inoculation would induce a long lasting immune response, four rabbits were inoculated once with ALA virus (3.5 x 10(5) PFU) and their serologic and mucosal antirotavirus titers were monitored at intervals for 1.5-2 years. The infected rabbits maintained serologic and mucosal rotavirus antibody titers until the final time point more than 700 days post inoculation. These data are important because they indicate that the antigenic load achieved following a single oral inoculation is sufficient to achieve long lasting immunity, the goal of any potential vaccine.

Animals↗