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Systemic and mucosal immune responses to rhesus rotavirus vaccine MMU 18006.

Thirty-four children 3 to 20 months of age ingested either 10(5), 10(4) or 10(3) plaque-forming units of rhesus rotavirus vaccine, MMU 18006, which possesses human rotavirus serotype 3 neutralization antigen. Immune responses were evaluated by a plaque reduction neutralization (PRN) assay to rotavirus serotypes 1, 2 and 3 and by a serum IgG, IgM and IgA and fecal IgA class-specific enzyme-linked immunosorbent assay. Homotypic PRN antibody seroconversions to serotype 3 rotavirus were detected in 31 of 34 children (91%), whereas rises in heterotypic PRN antibody to human rotavirus serotypes 1 or 2 were found in only 3 of 21 (14%) (p less than 0.00000001). Thirty of the 34 vaccinated children (88%) had at least one class of rotavirus-specific serum antibody detected by enzyme-linked immunosorbent assay. A rotavirus-specific IgA coproantibody response was seen in 11 of 16 children (69%) following vaccination. Two children who had no evidence of PRN antibody to serotype 3 after vaccination had evidence of both a fecal and a serum rotavirus-specific IgA response, suggesting that in these children the response to the vaccine was primarily mucosal. These data show that orally administered rhesus rotavirus vaccine MMU 18006 elicits local intestinal immunity but produces primarily a homotypic serum neutralization response as measured by plaque reduction neutralization assays.

Antibodies, Viral↗

Molecular epidemiology of human rotaviruses in Santiago, Chile.

BACKGROUND: Protective immunity against rotavirus infection is directed against antigenic epitopes on the outer capsid proteins VP7 and VP4. Our aim was to characterize the epidemiology of rotavirus antigenic types over time in Santiago, Chile. METHODS: We prospectively obtained 2097 stool samples for rotavirus testing, VP7 (G1 to G4) and VP4 (P4, P6, P8, P9) typing from children with diarrhea evaluated in emergency rooms of 5 base hospitals of Santiago. In addition 256 rotavirus-positive samples collected between 1985 and 1987 in the north health care area of Santiago were studied. RESULTS: Of 995 rotavirus-positive samples obtained 825 (82%) were typable for 1 or more VP7 types. G1 represented 81% of the G-typed samples during 1993 through 1995 and 77% during 1985 through 1987, predominating in all health care areas. G2 was next most common in all 5 areas, representing 6 to 23% of typed samples, with 1 area, the Southeast concentrating a significantly higher number of G2 infections. G2 declined from 35% of rotavirus-positive samples in 1993 to 0% in 1995 (P < 0.001), and from 25% to 2% in the north health care area from 1985 to 1987 (P < 0.001). G4 was uncommon and significantly more prevalent in 1985 through 1987 than in 1993 through 1995 (7% vs. 3%, P = 0.015). G3 was not detected. G1P8 (53%) and G2P4 (16%) combinations were by far the most commonly detected G-P associations. CONCLUSIONS: In Santiago, Chile, rotavirus antigenic type G1P8 has been highly prevalent and G2P4 has circulated in cycles. Differences in epidemiology of rotavirus antigenic types worldwide may prove to be relevant in efficacy of rotavirus vaccines.

Child↗

International variation in disease burden of rotavirus gastroenteritis in children with community- and nosocomially acquired infection.

BACKGROUND: The great impact of rotavirus disease on morbidity and medical health care costs in industrialized countries together with the withdrawal of the live oral rotavirus vaccine have made a reassessment of rotavirus gastroenteritis necessary. Such a reassessment should provide sufficient data for developing alternative disease prevention strategies and for allocating resources efficiently. OBJECTIVES: To compare characteristics and management of community- and nosocomially acquired rotavirus disease in Austria, Germany and Switzerland. PATIENTS AND METHODS: In a prospective, population-based, trinational (Austria, Germany, Switzerland), multicenter (9 cities, 10 hospitals and 30 pediatric practices) study, a total of 174 552 children months and 78 516 hospital days were evaluated. Participants were all children 4 years of age and younger, who either presented at one of the pediatric practices with community-acquired gastroenteritis, or who had acquired gastroenteritis nosocomially. From December, 1997, to May, 1998, prospective antigen testing was done by enzyme-linked immunosorbent assay, and serotyping was done by reverse transcription polymerase chain reaction. Disease severity was scored by the Vesikari severity scale. RESULTS: Rotavirus was detected in 29.5, 27 and 37.5% of children with community-acquired gastroenteritis and in 57, 69 and 49% of children with nosocomial gastroenteritis in Austria, Germany and Switzerland, respectively. Severity of community-acquired rotavirus gastroenteritis was more pronounced in Austria (median severity score, 11) than in Germany (median score, 9) or Switzerland (median score, 10). However, only 2% of Austrian and Swiss children compared with 12% of German children presented to their pediatricians more than four times. Nosocomially acquired rotavirus gastroenteritis was mildest in Austria but occurred within the shortest median duration of hospitalization (4 days vs. 5 and 7 in Germany and Switzerland, respectively). In a multivariant analysis age, family size, day care, breast-feeding and nationality were not predictive factors for enhanced risk to contract rotavirus infection. Alimentation was changed frequently; diet was used between 23 and 83%; special formulas were used between 10 and 57%. CONCLUSION: The cumulative experience from three European countries suggest that rotavirus is an important cause of diarrhea in Central Europe, but significant local differences clearly demonstrate the need for obtaining national data as a reliable basis for control and prevention of the disease.

Child, Preschool↗

A definition of bovine rotavirus virulence.

Rotaviruses are accepted as enteric pathogens of calves but many natural infections are subclinical. In the present paper, the outcome of inoculation of gnotobiotic calves of three ages (the second day of life, the second week of life and calves aged 6 weeks and over) with doses of 10(5.0) to 10(6.5) TCID50 was compared for three bovine rotavirus isolates (C3-160, 17/4 and 39/58). The clinical outcome of infection was dependent on both calf age and rotavirus isolate. Age-dependent resistance to infection was not found. By contrast, age-dependent resistance to disease was found with rotavirus isolates C3-160 and 17/4 but not with 39/58. All three isolates caused disease in calves inoculated on the second day of life but only one, 39/58, caused disease in the two older groups. Peak levels and duration of virus excretion were similar in clinically normal (10(6.7 +/- 1.08) TCID50 per g of faeces for 4.6 +/- 1.2 days) and diseased (10(7.45 +/- 0.94) TCID50 per g of faeces for 5.3 +/- 0.98 days) calves of all ages, but the onset of virus excretion occurred sooner in clinically affected calves (1.6 +/- 0.63 days in clinically affected compared with 3.7 +/- 1.5 days in clinically normal calves, P < 0.01). The present study confirmed the findings of an earlier study (Bridger & Pocock, 1986) which showed that bovine rotaviruses differ in virulence for calves in the second week of life and that older calves are susceptible to rotavirus infection and disease. In addition the present study demonstrated for the first time, that differences in rotavirus virulence are not apparent with calves inoculated on the second day of life, an age which has been used commonly to assess rotavirus virulence. It is suggested that rotaviruses that cause disease in calves only on the second day of life should be described as of low virulence whereas those that cause disease in all ages should be described as virulent.

Aging↗

Recognition of rotavirus antigens by mouse L3T4-positive T helper cells.

A lymphocyte proliferation assay was used to examine the helper T cell response to rotavirus in mice following parenteral immunization with the UK strain of bovine rotavirus. Mixed populations of lymphocytes prepared from spleen or peripheral lymph nodes were tested for proliferation in the presence of UK strain rotaviruses, prepared as cell culture lysates, ultracentrifuged (pelleted) lysates, sucrose-purified virus and caesium chloride-purified virus. Live rotavirus induced non-specific stimulation of lymphocytes, which was not observed in response to inactivated virus. Putative helper T cells of the L3T4+ phenotype were prepared as an enriched population from UK strain-immunized mice or grown in vitro as a polyclonal T cell line. The response of L3T4(+)-enriched cells from mice immunized with inactivated virus was dependent on antigen-presenting cells (APCs). Cells obtained following immunization with live virus did not require further addition of APCs. The response of the L3T4+ T cell line was wholly dependent on APCs. UK strain-specific L3T4+ cells responded to whole UK rotavirus and to isolated VP6 of both UK and C486 rotavirus strains. The results indicate that virus-specific L3T4+ T cells are induced following rotavirus immunization and can respond to epitopes on VP6. UK strain-primed L3T4+ cells also responded to an avian rotavirus strain, Ch2, which shares only minimal serological cross-reactivity with the UK strain. T cell recognition of rotavirus may thus be broadly cross-reactive.

Animals↗

Rotavirus spike protein VP5* binds alpha2beta1 integrin on the cell surface and competes with virus for cell binding and infectivity.

Rotaviruses recognize several cell-surface molecules, including the alpha2beta1 integrin, and the processes of rotavirus cell attachment and entry appear to be multifactorial. The VP5* subunit of the rotavirus spike protein VP4 contains the alpha2beta1 ligand sequence Asp-Gly-Glu at residues 308-310. Binding to alpha2beta1 and infectivity of monkey rotavirus strain RRV and human rotavirus strain Wa, but not porcine rotavirus strain CRW-8, are inhibited by peptides containing Asp-Gly-Glu. Asp308 and Gly309 are necessary for the binding of RRV VP5* (aa 248-474) to expressed I domain of the alpha2 integrin subunit. Here, the ability of RRV VP5* to bind cells and affect rotavirus-integrin interactions was determined. Interestingly, VP5* bound to cells at 4 and 37 degrees C, both via alpha2beta1 and independently of this integrin. Prior VP5* binding at 37 degrees C eliminated RRV binding to cellular alpha2beta1 and reduced RRV and Wa infectivity in MA104 cells by 38-46 %. VP5* binding did not affect the infectivity of CRW-8. VP5* binding at 4 degrees C did not affect permissive-cell infection by RRV, indicating an energy requirement for VP5* competition with virus for infectivity. Mutagenesis of VP5* Asp308 and Gly309 eliminated VP5* binding to alpha2beta1 and the VP5* inhibition of rotavirus cell binding and infection, but not alpha2beta1-independent cell binding by VP5*. These studies show for the first time that expressed VP5* binds cell-surface alpha2beta1 using Asp308 and Gly309 and inhibits the infection of homologous and heterologous rotaviruses that use alpha2beta1 as a receptor.

Animals↗

Rotavirus hospitalisation in New Zealand children under 3 years of age.

OBJECTIVE: To describe the epidemiology of severe rotavirus gastroenteritis and to estimate the hospitalisation rates of this illness in New Zealand children under 3 years of age. METHODS: Children under 3 years of age with acute diarrhoea admitted to 1 of 8 study hospitals between 1 May 1998 and 30 April 2000 were surveyed. Their socio-demographic, treatment and length-of-stay data were recorded and stool samples tested by a rotavirus-specific enzyme-linked immunoassay. National hospital discharge data for infectious diarrhoea (International Classification of Diseases, ninth revision, 003-009) were reviewed, allowing population-based estimates for rotavirus-related hospitalisation in New Zealand. RESULTS: Of 2019 enrolled children, 1138 (56.4%) provided stools for testing, and of these 485 (42.6%) tested rotavirus positive. Rotavirus detection varied significantly by age (26.8% for 0 to 5 months, 42.5% for 6 to 11 months and 52.1% for children aged 12 to 35 months; P < 0.001), and by season (51.2% in winter/spring vs. 24.5% in summer/autumn; P < 0.001). While those infected with rotavirus were more likely to be dehydrated (50.6% vs. 37.4%; P < 0.001), their median hospital stay was similar (1.0 vs. 2.0 days; P = 0.09) to other children with acute gastroenteritis. The estimated national hospitalisation rate for rotavirus diarrhoea in children under 3 years, standardised for age and season, was 634 (95% CI 597, 672) per 100,000. In New Zealand, rotaviruses result in 1 in 52 children being hospitalised by 3 years of age. CONCLUSIONS: Rotavirus diarrhoea is an important, potentially vaccine-preventable cause of hospitalisation in New Zealand children, especially during winter and spring seasons.

Age Distribution↗

Rotavirus and other enteropathogens in childhood acute diarrhoea: a study of two centres in Malaysia.

AIM: To study the role of rotavirus in children hospitalised for acute gastroenteritis (AGE) in two urban hospitals in Malaysia. METHODS: A 12-month prospective study (January to December 2002), in children younger than 14 years with AGE hospitalised to the paediatric units of University of Malaya Medical Centre (UMMC), Kuala Lumpur; and Hospital Sultanah Aminah (HSA), Johor Bahru, Malaysia was conducted. RESULTS: In 2002, 399 and 1307 children with AGE were admitted to UMMC and HSA, respectively. Two hundred and eighty-eight (72%) stool samples from UMMC and 901 (69%) samples from HSA were analysed. Rotavirus was the most common aetiological agent identified in both centres (average 32%; UMMC 35%, HSA 30%, P = 0.94). The peak age group for rotavirus-related hospitalisation was 24-35 months for UMMC and 12-23 months for HSA. Nine percent of patients hospitalised for rotavirus infection in UMMC and 22% of patients in HSA were older than 5 years of age. An outbreak of rotavirus infection within the communities served by both centres resulting in an increase in hospital admissions of rotavirus gastroenteritis was observed in both units from January to March 2002. CONCLUSION: The peak age group for rotavirus-related hospital admission in this study was much older, between 12 to 35 months. It is uncertain whether this was related to the outbreak of rotavirus gastroenteritis observed within two urban areas from January to March 2002 causing re-infection with rotavirus in older children.

Acute Disease↗

Rotavirus vaccine studies in Europe.

Following the discovery and recognition of rotavirus as an important human enteropathogen, work towards the development of rotavirus vaccines was begun in the USA and Europe. The first candidate rotavirus vaccine was launched by SmithKline-RIT, Belgium, and studied in clinical trials in Finland, Yugoslavia, Italy, Switzerland, Austria and the UK. Efficacy trials in Finland of RIT 4237 strain live oral bovine rotavirus vaccine resulted in some fundamental findings about rotavirus immunity in humans: protection against disease rather than infection; better protection against severe than mild disease and heterotypic rather than homotypic protection. Another bovine rotavirus vaccine, strain WC-3, was studied briefly in France. Also studies of rhesus rotavirus vaccine were started early in Europe, and efficacy studies were carried out in Finland and Sweden. Recently, rhesus-human re-assortant tetravalent (RRV-TV) rotavirus vaccine was tested in a field trial in Finland this study was pivotal for the registration of the vaccine in the USA and European Union countries. Despite extensive experience with rotavirus vaccines in Europe, the need for such vaccines in many European countries still requires further assessment.

Animals↗

Seasonality and diversity of Group A rotaviruses in Europe.

Group A rotaviruses are a major cause of severe gastroenteritis in children under 4 y of age worldwide. Group A rotaviruses have been identified in many animal and bird species, they are antigenically complex, and multiple serotypes infect humans. Re-assortant rotavirus vaccines are now available which confer protection against severe illness due to rotavirus serotypes G1-4. Before vaccines are introduced it is necessary to establish the diversity of rotavirus in the target population to ensure efficacy and to establish a baseline for future surveillance strategies. The purpose of this review is to describe our current knowledge of the diversity of rotaviruses across Europe. Since multinational studies with standardized methodology have not been performed, this review is based on the available published studies. In Europe, more than 90% of Group A rotavirus strains that have been typed are of serotypes G1-4, with an average 8% of non-G1-4 strains in published studies. The percentage of non-typeable strains may fluctuate from one year to another, and has been as high as 18% in one study in Great Britain, indicating the need for a more systematic study. Group A rotavirus infection typically occurs as a winter peak in the European countries studied. Comparison of seasonality data from national laboratory surveillance systems showed seasonal differences, with the annual rotavirus peak occurring first in Spain, usually in December, followed by France in February, and ending in Northern Europe in England and Wales in February or March, and the Netherlands and Finland in March.

Child↗

Rotavirus gastroenteritis in Finland: burden of disease and epidemiological features.

The burden of disease attributable to childhood rotavirus infection in Finland was assessed from data on hospital admissions for acute gastroenteritis and from reported virological diagnoses of rotavirus from 1985 to 1995. The mean number of hospitalizations (3584 annually in children under 5 y of age) corresponded to approximately 5.6% of the birth cohort. Rotavirus was estimated to be responsible for 54% of cases; accordingly, 3% of all children in Finland are hospitalized for rotavirus diarrhoea. The monthly distribution of hospitalizations for acute diarrhoea showed a similar pattern as monthly diagnoses of rotavirus, with a long epidemic period starting as early as November or December and lasting until June or even July. The prevalent rotavirus G-type throughout the study period was G1, which was detected in over 60% of the cases; however, in the season 1988-89 G4 was the prominent type. Improved case management has led to a shorter duration of hospital stay (3.3 d in 1985 vs. 2.3 d in 1995), but otherwise these was no significant trend for rotavirus gastroenteritis over the years. These findings underscore the need to control rotavirus gastroenteritis with a specific intervention, notably rotavirus vaccination.

Acute Disease↗

Rotavirus vaccines: an overview.

Rotavirus vaccine development has focused on the delivery of live attenuated rotavirus strains by the oral route. The initial "Jennerian" approach involving bovine (RIT4237, WC3) or rhesus (RRV) rotavirus vaccine candidates showed that these vaccines were safe, well tolerated, and immunogenic but induced highly variable rates of protection against rotavirus diarrhea. The goal of a rotavirus vaccine is to prevent severe illness that can lead to dehydration in infants and young children in both developed and developing countries. These studies led to the concept that a multivalent vaccine that represented each of the four epidemiologically important VP7 serotypes might be necessary to induce protection in young infants, the target population for vaccination. Human-animal rotavirus reassortants whose gene encoding VP7 was derived from their human rotavirus parent but whose remaining genes were derived from the animal rotavirus parent were developed as vaccine candidates. The greatest experience with a multivalent vaccine to date has been gained with the quadrivalent preparation containing RRV (VP7 serotype 3) and human-RRV reassortants of VP7 serotype 1, 2, and 4 specificity. Preliminary efficacy trial results in the United States have been promising, whereas a study in Peru has shown only limited protection. Human-bovine reassortant vaccines, including a candidate that contains the VP4 gene of a human rotavirus (VP4 serotype 1A), are also being studied.

Animals↗

Passive immunity to bovine rotavirus in newborn calves fed colostrum supplements from immunized or nonimmunized cows.

Colostrum was collected and pooled from each of five cows in three experimental groups: group I cows received intramuscular and intramammary inoculations of adjuvanted modified live Ohio Agricultural Research and Development Center rotavirus vaccine; group II cows were injected intramuscularly with a commercial modified-live rota-coronavirus vaccine; and group III cows were uninoculated controls. Pooled colostrum from group I cows had higher (P less than 0.05) enzyme-linked immunosorbent assay (ELISA) immunoglobulin G (IgG1) and virus neutralization (VN) rotavirus antibody titers (ELISA IgG1 = 2,413,682; VN = 360,205) than did colostrum from group II (ELISA IgG1 = 8,192; VN = 4,395) or group III cows (ELISA IgG1 = 5,916; VN = 2,865). The antibody titers of these last two colostrum pools did not differ (P greater than 0.05). Samples of these colostrum pools were fed as daily supplements (percent [vol/vol] in cow's milk infant formula) to 28 newborn, unsuckled, antibody-seronegative, male Holstein calves. Eight calves received no supplemental colostrum. The calves were orally challenged with virulent bovine rotavirus and monitored daily for diarrhea and fecal rotavirus shedding. Diarrhea and rotavirus shedding occurred in the eight calves fed no supplemental colostrum and persisted longest in this group. The pooled colostrum from group I cows protected eight of eight calves from both rotavirus diarrhea and shedding when fed as a 1% supplement. The pooled colostrum from neither group II nor group III cows protected 12 other calves against rotavirus diarrhea or shedding when fed at the same concentration (1%). Six rotavirus-challenged calves fed 0.1% supplemental colostrum from group I cows and two calves fed 10 and 50% supplemental colostrum from control cows displayed partial passive immunity, exemplified by delayed onset and shortened duration of rotavirus-associated diarrhea and virus shedding.

Animals↗

Molecular epidemiology of G9 rotaviruses in Taiwan between 2000 and 2002.

Since the mid-1990s, novel G9 rotaviruses have been detected in many countries, suggesting that G9 is a globally important serotype. The molecular epidemiology of G9 rotaviruses in Taiwan from 2000 to 2002 was investigated in this study. G9 rotavirus first appeared in 2000 with 4 cases and constituted 33.8% and 54.8% of the rotavirus-positive samples in 2001 and 2002, respectively. These G9 strains belonged to P[8]G9, subgroup II, and long electropherotype, except one belonged to P[4]G9, subgroup II, and short electropherotype. Nucleotide sequencing and phylogenetic analysis of 52 Taiwanese G9 rotaviruses showed that the VP7 genes shared a high degree of identity to overseas G9 rotaviruses detected after 1993 and that the VP8* portions of the VP4 genes were more closely related to those of local rotaviruses of other G types. The two P[8]G9 strains with high nucleotide identities in the VP7 and the partial VP4 genes, 01TW591 of Taiwan from 2001 and 95H115 of Japan from 1995, varied in four genes, genes 2, 3, 7, and 8, which was revealed by RNA-RNA hybridization. Representative strains for different RNA patterns were also analyzed in the partial VP2 and VP3 genes; the nucleotide identities were high between Taiwanese G9 strains and local G3 or G2 strains. These results suggested that Taiwanese G9 rotaviruses possibly had evolved through reassortment between overseas G9 strains and circulating rotaviruses of other G types.

Antigens, Viral↗

Electron microscopy procedure influences detection of rotaviruses.

Technical parameters of electron microscope staining procedures (type of stain, pH of stain, and time of staining) influence particle integrity for three groups of rotaviruses. Simian rotavirus SA11 (group A), Chinese adult diarrhea rotavirus and porcine rotavirus-like agent (group B), and porcine pararotavirus (group C) were tested. All rotavirus strains were quite stable in uranyl acetate and phosphotungstic acid at pH 4.5 and relatively stable in ammonium molybdate. However, staining with phosphotungstic acid at higher pH values with increased staining time yielded a reduction in the number of particles and particles that were broken or degraded to single-shelled particles or core particles. The different staining procedures were also tested in immunoelectron microscopy experiments. Antibody molecules bound to rotavirus particles were observed clearly only with phosphotungstic acid staining and not with uranyl acetate. We therefore recommend that uranyl acetate and phosphotungstic acid at pH 4.5 be used for negative staining of rotaviruses; phosphotungstic acid at pH 4.5 is optimal for immunoelectron microscopy. These technical points may be critical for rotavirus detection and are important for studies pertaining to the epidemiology and clinical importance of the non-group A rotaviruses.

Animals↗

Role of coproantibody in clinical protection of children during reinfection with rotavirus.

Rotavirus is the major cause of severe, dehydrating infantile gastroenteritis. Infection is limited to the gut, but the relative roles of serum and secretory copro-immunoglobulin A (IgA) in protection are unclear. Specific copro-IgA is predictive of duodenal antirotaviral IgA and correlates with virus-neutralizing coproantibody. Copro-IgA conversion is a more sensitive marker of rotavirus reinfection than seroconversion. We measured rotavirus reinfections by copro-IgA conversion prospectively in 35 children recruited at a time of severe rotavirus illness. The children were followed up longitudinally for 14 to 31 months to determine whether high coproantibody levels correlated with clinical protection against rotavirus disease. Ninety-four percent of the children experienced reinfection, and 38% developed persistent elevations in specific copro-IgA termed plateaus. Plateau children had a higher mean annual rate of rotavirus infection and a lower ratio of symptomatic to total number of rotavirus reinfections than did nonplateau children. The annual rates of rotavirus infection and disease were significantly higher outside the plateau than inside it in children experiencing antirotavirus copro-IgA plateaus. Frequent rotavirus infection of children appears to stimulate production of a specific copro-IgA plateau which correlates with protection against an excess of infection and symptomatic disease.

Age Factors↗

Detection of rotavirus serotypes G1, G2, G3, and G11 in feces of diarrheic calves by using polymerase chain reaction-derived cDNA probes.

On the basis of antigenic variability in the VP7 outer capsid glycoprotein, at least 14 G serotypes exist for group A rotaviruses. Serotypic diversity exists among bovine rotaviruses (BRV), with serotypes G1, G6, G8, and G10 reported for cattle. Although G1 and G8 rotaviruses were originally described for humans, the recent isolation of G6 and G10 rotaviruses from humans further emphasizes the serotypic similarity between human and bovine rotaviruses and the possible zoonotic potential of rotaviruses. Results of our previous studies have indicated that more than 24% of BRV-positive field samples from diarrheic calves were nonreactive with cDNA probes or monoclonal antibodies to serotypes G6, G8, and G10. In this study, cDNA probes were prepared by polymerase chain reaction amplification of the hyperdivergent regions of the VP7 genes (nucleotides 51 to 392) from human (G1, G2, and G3) and porcine (G4, G5, and G11) rotaviruses. These probes were used in a dot blot hybridization assay to further characterize the G types of 59 BRV strains (fecal samples from diarrheic calves in Ohio, Nebraska, Washington, and South Dakota) that were nonreactive with cDNA probes to G6, G8, and G10. Rotaviruses belonging to serotypes G1 (n = 7), G2 (n = 1), G3 (n = 2), and G11 (n = 3) were identified among the BRV field samples. The BRV associated with these G types accounted for 22% of the samples tested; the other 78% of these samples remained untypeable with these probes. To our knowledge, this is the first report in the United States of the identification among BRV isolates of rotavirus serotypes G1, G2, G3, and G11.

Animals↗

Serotypic differentiation of rotaviruses in field samples from diarrheic pigs by using nucleic acid probes specific for porcine VP4 and human and porcine VP7 genes.

Of 216 fecal and intestinal samples collected from nursing or weaned diarrheic pigs in the United States and Canada, 57 were identified as group A rotavirus positive by RNA electrophoresis and silver staining. Fifty-seven and 52 rotavirus-positive samples were analyzed by hybridization with Gottfried and OSU PCR-derived gene 9 and 4 probes, respectively. Only 17 samples were identified with either homologous VP4 (P)- or VP7 (G)-coding genes or both. One rotavirus identified as G4 and P7 was similar to the previously characterized interserotype rotavirus, SB-1A. Additional hybridization analyses were performed with PCR-derived probes prepared from gene 9 cDNA of the human rotaviruses Wa (G1), DS-1 (G2), and P (G3) and of the porcine rotavirus YM (G11). Eleven of 52 samples collected and analyzed from swine in Ohio, California, and Nebraska were identified as G11. No samples with G1-, G2-, or G3-type specificities were detected among the 25 of 57 rotavirus-positive samples analyzed with human rotavirus-derived probes. Further investigations with a PCR-derived gene 4 probe prepared from porcine rotavirus YM revealed hybridization specificities similar to those of the OSU gene 4 probe.

Animals↗