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PubMed1987

Evaluation of new commercial enzyme immunoassay for rotavirus detection.

We evaluated a new commercial enzyme immunoassay (EIA) for rotavirus (Rotavirus EIA; International Diagnostic Laboratories, Chesterfield, Mo.). A total of 161 consecutive stool samples (including 18 from infants less than 30 days old) submitted to the diagnostic laboratory at Children's Hospital, Washington University Medical Center, St. Louis, Mo., for rotavirus detection were tested by Rotavirus EIA and by Rotazyme II (Abbott Laboratories, North Chicago, III.) according to the instructions of the manufacturer. In addition, 16 samples from infants less than 30 days old without diarrhea were tested by both assays. Samples showing discrepant results after repeat testing were examined by electron microscopy. Nine samples yielding discrepant results were also tested by using a reference EIA directly on the specimen and on culture supernatants from two passages in MA 104 cells. Rotavirus EIA and Rotazyme II yielded concordant results for 85% of the samples. All of the 26 discrepant samples tested negative by Rotavirus EIA and positive (15 samples) or equivocal (11 samples) by Rotazyme II. These samples included 11 from symptomatic infants more than 30 days old, 2 from symptomatic infants less than 30 days old (neonates), and 2 from neonates without diarrhea. Rotavirus was not detected in any of the 24 that were examined by electron microscopy or in any of the 9 that were tested by the reference EIA. The sensitivity, specificity, positive predictive value, and negative predictive value were 100% for Rotazyme EIA and 100, 90, 70, and 100%, respectively, for Rotazyme II. Rotavirus EIA was comparable to Rotazyme II in ease of performance. We conclude that Rotavirus EIA is equally sensitive and more specific than Rotazyme II for detecting rotavirus. Rotavirus EIA is a practical and accurate rotavirus assay for use in clinical laboratories.

Diarrhea, Infantile↗
PubMed1987

Genetic relatedness among human rotavirus genes coding for VP7, a major neutralization protein, and its application to serotype identification.

Antigenic characterization of human rotaviruses by plaque reduction neutralization assay has revealed four distinct serotypes. The outer capsid protein VP7, coded for by gene 8 or 9, is a major neutralization protein; however, studies of rotaviruses derived from genetic reassortment between two strains have confirmed that another outer capsid protein, VP3, is in some cases equally important in neutralization. In this study, the genetic relatedness of the genes coding for VP7 of human rotaviruses belonging to serotypes 1 through 4 was examined by hybridization of their denatured double-stranded genomic RNAs to labeled single-stranded mRNA probes derived from human-animal rotavirus reassortants containing only the VP7 gene of their human rotavirus parent. A high degree of homology was demonstrated between the VP7 genes of strain D and other serotype 1 human rotaviruses, strain DS-1 and other serotype 2 human rotaviruses, strain P and other serotype 3 human rotaviruses, and strain ST3 and other serotype 4 human rotaviruses. Hybrid bands could not be demonstrated between the VP7 gene of D, DS-1, P, or ST3 and the corresponding gene of human rotaviruses belonging to a different serotype. RNA specimens extracted from the stools of 15 Venezuelan children hospitalized with rotavirus diarrhea were hybridized to each of the reassortant probes representing the four human serotypes. All five viruses with short RNA patterns showed homology with the DS-1 strain VP7 gene; two of these were previously adapted to tissue culture and shown to be serotype 2 strains by tissue culture neutralization. Of the remaining 10 viruses with long RNA patterns, 2 hybridized only to the D strain VP7 gene, 6 hybridized only to the P strain VP7 gene, and 2 hybridized only to the ST3 strain VP7 gene. Hybridization using single human rotavirus gene substitution reassortants as probes may provide an alternative method for identifying the VP7 serotype of field isolates that would circumvent the need for tissue culture adaptation.

Capsid↗
PubMed1999

Antibody-independent protection against rotavirus infection of mice stimulated by intranasal immunization with chimeric VP4 or VP6 protein.

This study was to determine whether individual rotavirus capsid proteins could stimulate protection against rotavirus shedding in an adult mouse model. BALB/c mice were intranasally or intramuscularly administered purified Escherichia coli-expressed murine rotavirus strain EDIM VP4, VP6, or truncated VP7 (TrVP7) protein fused to the 42.7-kDa maltose-binding protein (MBP). One month after the last immunization, mice were challenged with EDIM and shedding of rotavirus antigen was measured. When three 9-microg doses of one of the three rotavirus proteins fused to MBP were administered intramuscularly with the saponin adjuvant QS-21, serum rotavirus immunoglobulin G (IgG) was induced by each protein. Following EDIM challenge, shedding was significantly (P = 0.02) reduced (i.e., 38%) in MBP::VP6-immunized mice only. Three 9-micrograms doses of chimeric MBP::VP6 or MBP::TrVP7 administered intranasally with attenuated E. coli heat-labile toxin LT(R192G) also induced serum rotavirus IgG, but MBP::VP4 immunization stimulated no detectable rotavirus antibody. No protection against EDIM shedding was observed in the MBP::TrVP7-immunized mice. However, shedding was reduced 93 to 100% following MBP::VP6 inoculation and 56% following MBP::VP4 immunization relative to that of controls (P = <0.001). Substitution of cholera toxin for LT(R192G) as the adjuvant, reduction of the number of doses to 1, and challenge of the mice 3 months after the last immunization did not reduce the level of protection stimulated by intranasal administration of MBP::VP6. When MBP::VP6 was administered intranasally to B-cell-deficient microMt mice that made no rotavirus antibody, shedding was still reduced to <1% of that of controls. These results show that mice can be protected against rotavirus shedding by intranasal administration of individual rotavirus proteins and that this protection can occur independently of rotavirus antibody.

ATP-Binding Cassette Transporters↗
PubMed2004

Inhibition of cyclooxygenase activity reduces rotavirus infection at a postbinding step.

Elevated levels of prostaglandins (PGs), products of cyclooxygenases (COXs), are found in the plasma and stool of rotavirus-infected children. We sought to determine the role of COXs, PGs, and the signal transduction pathways involved in rotavirus infection to elucidate possible new targets for antiviral therapy. Human intestinal Caco-2 cells were infected with human rotavirus Wa or simian rotavirus SA-11. COX-2 mRNA expression and secreted PGE2 levels were determined at different time points postinfection, and the effect of COX inhibitors on rotavirus infection was studied by an immunofluorescence assay (IFA). To reveal the signal transduction pathways involved, the effect of MEK, protein kinase A (PKA), p38 mitogen-activated protein kinase (MAPK), and NF-kappaB inhibitors on rotavirus infection was analyzed. In infected Caco-2 cells, increased COX-2 mRNA expression and secreted PGE2 levels were detected. Indomethacin (inhibiting both COX-1 and COX-2) and specific COX-1 and COX-2 inhibitors reduced rotavirus infection by 85 and 50%, respectively, as measured by an IFA. Indomethacin reduced virus infection at a postbinding step early in the infection cycle, inhibiting virus protein synthesis. Indomethacin did not seem to affect viral RNA synthesis. Inhibitors of MEK, PKA, p38 MAPK, and NF-kappaB decreased rotavirus infection by at least 40%. PGE2 counteracted the effect of the COX and PKA inhibitors but not of the MEK, p38 MAPK, and NF-kappaB inhibitors. Conclusively, COXs and PGE2 are important mediators of rotavirus infection at a postbinding step. The ERK1/2 pathway mediated by PKA is involved in COX induction by rotavirus infection. MAPK and NF-kappaB pathways are involved in rotavirus infection but in a PGE2-independent manner. This report offers new perspectives in the search for therapeutic agents in treatment of severe rotavirus-mediated diarrhea in children.

Antiviral Agents↗
PubMed2006

Molecular characterisation of the rotavirus strains prevalent in Maua, Meru North, Kenya.

BACKGROUND: Rotavirus is the most common cause of severe infantile diarrhoea disease in infants and young children below five years worldwide. Rotavirus is associated with high cases of morbidity and mortality and it is estimated that up to 650,000 deaths in young children occur annually in the less developed countries and approximately 150,000-200,000 deaths occur in Africa alone. OBJECTIVE: To characterise the circulating rotavirus strains in Maua, Meru North district, Kenya. DESIGN: A prospective study to investigate and characterise rotavirus serotypes/genotypes and electropherotypes in infants and children with severe diarrhoea hospitalised and/or attending the outpatient department of Maua Methodist Hospital during the period April 2004 to September 2005. SETTING: Maua Methodist Hospital, Meru North, Kenya. SUBJECTS: Faecal samples were collected from 135 infants and children with acute diarrhoea and were screened first for the presence of human Group A rotavirus antigen using commercially available enzyme linked immunosorbent assay kit (ELISA). The positive samples were evaluated by sodium dodecyl polycrylamide gel electrophoresis (SDS-PAGE) to determine the viral RNA electropherotype profile. Rotavirus strains were also genotyped using reverse transcriptase polymerase chain reaction (RT-PCR) of the VP7 gene. RESULTS: Assay of these samples with commercial ELISA showed that 17.8% (24/135) were positive for group A rotavirus antigen. Twenty of these ELISA positive samples were also analysed by SDS-PAGE of which 75% (15/20) gave detectable electropherotype pattern with the long electropherotype being predominant 80.0% (12/15) followed by the short RNA profile 20.0% (2/ 15). Seventeen of the ELISA positive samples were genotyped for VP7 and the results showed that G9 was the most predominant genotype comprising 47.1% (8/17) followed by G8 29.4% (5/17), GI 17.4% (3/17) and the mixed genotype was G8/G9 5.9% (1/17). Most patients with rotavirus infection were of the age of 3 - 60 months, with 79% being less than 18 months old. CONCLUSION: The overall prevalence of rotavirus infection in young children with diarrhoea hospitalised and/or attending the out-patient department of Maua Methodist Hospital was 17.8% with the predominant serotype being G9. These results show that rotavirus plays an important role in severe viral diarrhoea in young children in Maua Meru North district, Kenya. Furthermore, this high G9 rotavirus prevalence in Kenya may require vaccine trials to be held in Kenya so as to determine the efficacy of new rotavirus vaccine candidates that do not include the G9 serotype.

Child, Preschool↗
PubMed1999

The epithelial cell response to rotavirus infection.

Rotavirus is the most important worldwide cause of severe gastroenteritis in infants and young children. Intestinal epithelial cells are the principal targets of rotavirus infection, but the response of enterocytes to rotavirus infection is largely unknown. We determined that rotavirus infection of HT-29 intestinal epithelial cells results in prompt activation of NF-kappaB (<2 h), STAT1, and ISG F3 (3 h). Genetically inactivated rotavirus and virus-like particles assembled from baculovirus-expressed viral proteins also activated NF-kappaB. Rotavirus infection of HT-29 cells induced mRNA for several C-C and C-X-C chemokines as well as IFNs and GM-CSF. Mice infected with simian rotavirus or murine rotavirus responded similarly with the enhanced expression of a profile of C-C and C-X-C chemokines. The rotavirus-stimulated increase in chemokine mRNA was undiminished in mice lacking mast cells or lymphocytes. Rotavirus induced chemokines only in mice <15 days of age despite documented infection in older mice. Macrophage inflammatory protein-1beta and IFN-stimulated protein 10 mRNA responses occurred, but were reduced in p50-/- mice. Macrophage inflammatory protein-1beta expression during rotavirus infection localized to the intestinal epithelial cell in murine intestine. These results show that the intestinal epithelial cell is an active component of the host response to rotavirus infection.

Aging↗
PubMed1994

Epidemiology, subgroups and serotypes of rotavirus diarrhea in north Indian communities.

To know prevalence of rotavirus diarrhea subgroups and serotypes, a prospective study was conducted in rural, periurban and urban communities at Chandigarh. Weekly surveillance for diarrheal episodes was carried out in 110 families each from rural, periurban and urban localities constituting 584 children < 5 years of age from October, 1988 to February, 1991. Stool samples of 218 diarrheal episodes occurring in 115 children were subjected to rotavirus detection by ELISA. Rotavirus positive samples were further analyzed for subgroups and serotypes using specific monoclonal antibodies. Overall prevalence of rotavirus diarrhea was 4.3% (25/584). Rotavirus constituted 11.5% (25/218) of total diarrheal episodes and 22% (25/115) among the children affected with acute diarrhea. Among rural, periurban and urban communities, the overall prevalences of rotavirus diarrhea were 7.3%, 3.2% and 2.3% and episode related prevalences of 31.8%, 7.4% and 5%, respectively (chi 2 test for trend was highly significant from rural to periurban to urban localities). Forty per cent (10/25 of rotavirus positive samples were subgroup I and 60% (15/25) sub-group II. Of the 25 rotavirus strains, 40% (10) were serotype 2, 24% (n = 6) serotype 3 and 36% (n = 9) serotype 4. No definite temporal or seasonal pattern of rotavirus was observed; however, more of rotavirus diarrheal episodes (16%) occurred during winter season. Subgroups and serotypes were observed to cocirculate during the rotavirus episodes. Demonstration of serotypes in our field study imply that the vaccine to be used in our country must be cross protective to have an effective impact on rotavirus infection.(ABSTRACT TRUNCATED AT 250 WORDS)

Child, Preschool↗
PubMed2003

Oral immunoglobulin for the prevention of rotavirus infection in low birth weight infants.

BACKGROUND: Rotavirus infection is the most common neonatal nosocomial viral infection. Epidemics with the newer P(6)G9 strains have been reported in neonatal units worldwide. These strains can cause severe symptoms in infected infants. Infection control measures become necessary and the utilisation of hospital resources increase. Local mucosal immunity in the intestine to rotavirus is important in the resolution of infection and protection against subsequent infections. Boosting local immunity by oral administration of anti-rotaviral immunoglobulin preparations might be a useful strategy in preventing rotaviral infections, especially in low birth weight babies. OBJECTIVES: To determine the effectiveness and safety of oral immunoglobulin preparations for the prevention of rotavirus infection in hospitalised low birthweight infants (birth weight less than 2500 gms) SEARCH STRATEGY: The Cochrane Central Register of Controlled Trials (CENTRAL, The Cochrane Library, Issue 2, 2002), MEDLINE, EMBASE and CINAHL were searched. Science Citation Index was searched for all articles which referenced Barnes 1982. The proceedings of the Pediatric Academic Societies which were published in the journal, Pediatric Research from 1991 onwards were searched as well as abstracts of doctoral dissertations and theses from 1960 onwards. The above mentioned search strategy was completed in June 2002. Authors prominent in the field were contacted for any unpublished articles and more information on published articles was sought. Reference lists of identified clinical trials and personal files were reviewed. SELECTION CRITERIA: The criteria used to select studies for inclusion were: 1) DESIGN: randomised or quasi-randomised controlled trials. 2) PARTICIPANTS: Hospitalised low birthweight infants. 3) INTERVENTION: Oral immunoglobulin preparations for prevention of rotavirus infection compared to placebo OR no intervention. 4) At least one of the following outcomes were reported: All cause mortality during hospital stay, mortality due to rotavirus infection during hospital stay, rotavirus infection, duration of diarrhoea, need for rehydration, duration of viral excretion, duration of infection control measures, length of hospital stay in days, recurrent diarrhoea or chronic diarrhoea. DATA COLLECTION AND ANALYSIS: The two reviewers independently abstracted data from the included trials MAIN RESULTS: One published study (Barnes 1982) was eligible for inclusion in this review. Two additional studies are awaiting assessment re eligibility for inclusion. Barnes 1982 found no significant difference in the rates of rotavirus infection after oral gammaglobulin versus placebo in hospitalised low birthweight babies [RR 1.27 (95% CI 0.65-2.37)]. In the subset of infants who became infected with rotavirus after receiving gammaglobulin or placebo for prevention of rotavirus infection, there was no significant difference in the duration of rotavirus excretion between the group who had gammaglobulin (mean 2 days, range 1-4 days) and the group who had placebo (mean 3 days, range 1-6 days). No adverse effects were reported by Barnes 1982 after administration of oral immunoglobulin preparations. REVIEWER'S CONCLUSIONS: Current evidence from one randomised controlled trial does not support the routine use of oral immunoglobulin preparations for the prevention of rotavirus infection in low birth-weight infants. However, newer immunoglobulin preparations which have been found to be effective in older children have not been tested in neonatal trials. Therefore, researchers should be encouraged to conduct well designed trials in neonates at risk for rotavirus infections using the newer preparations of anti-rotaviral immunoglobulins (colostrum, egg yolk immunoglobulins). Such trials should also include cost effectiveness evaluations.

Administration, Oral↗
PubMed1989

Induction and persistence of local rotavirus antibodies in relation to serum antibodies.

The induction and persistence of local rotavirus antibodies, including stool IgA, jejunal IgA, and jejunal neutralizing antibody, were evaluated in 14 adult volunteers infected with the CJN strain of human rotavirus. In addition, the relationships between local rotavirus IgA and serum rotavirus IgA, IgG, and neutralizing antibody were determined. Both stool and serum rotavirus IgA appeared to have similar kinetics. Both antibodies peaked by days 14-17 after inoculation in all subjects, then decreased rapidly. By days 26-28, titers had fallen to 13% and 30% of their respective peaks. Serum rotavirus IgG peaked somewhat later, occurring in five subjects on days 26-28. Serum neutralizing antibody peaked on days 26-28 in all but three subjects. Both serum IgG and neutralizing antibodies also declined more slowly than rotavirus IgA. Although all antibody concentrations had decreased to only a fraction of their peak responses by days 270-365 after rotavirus inoculation they remained higher than baseline levels. For example, stool rotavirus IgA concentrations were 13.5-fold higher than baseline, while jejunal rotavirus IgA and neutralizing antibody were 8.9- and 4.3-fold above baseline, respectively. Similarly, serum antibodies remained 3.7- to 11.2-fold higher than baseline at 270-365 days after rotavirus inoculation. These studies imply that serum rotavirus IgA is a good indicator of local antibody responses. Furthermore, although both serum and local antibody titers peaked within 2-4 weeks after infection, these antibodies persisted at above baseline concentrations for at least 9-12 months after infection.

Adolescent↗
PubMed1980

Chronic rotavirus infection in immunodeficiency.

The characteristics of rotavirus infection in 23 children with a variety of primary immunodeficiency diseases were studied. Stools and sera were tested for rotavirus by means of the enzyme-linked immunosorbent assay and the enzyme-linked fluorescent assay, respectively. Four immunodeficient patients had diarrhea during the study period and all had rotavirus infection; rotavirus was not detected in the stools of the 19 asymptomatic immunodeficient patients. Forty-six control children with diarrhea were tested and 22 had rotavirus infection; rotavirus was not detected in 39 asymptomatic control children. One immunodeficient patient with X-linked agammaglobulinemia and one with severe combined immunodeficiency had chronic, symptomatic rotavirus infection with rotavirus excretion lasting more than six weeks. The other two immunodeficient patients and eight control children eliminated the rotavirus from their stools in periods ranging from two to 12 days. Rotavirus antigen was detected in the sera of three of the four immunodeficient patients; none of the 14 control infants tested had rotavirus antigen detected in their sera. This study indicates that rotavirus may produce a chronic infection in immunodeficient children.

Adolescent↗
PubMed2001

Effect of concomitant HIV infection on presentation and outcome of rotavirus gastroenteritis in Malawian children.

BACKGROUND: Rotaviruses represent important causes of severe diarrhoea in early childhood. We examined the effect of HIV infection on the presentation and outcome of rotavirus gastroenteritis in Malawian children. METHODS: Children younger than 5 years who were treated for acute gastroenteritis at the Queen Elizabeth Central Hospital in Blantyre from July, 1997, to June, 1999, were enrolled. Children with rotavirus diarrhoea, with and without HIV infection, were followed up for up to 4 weeks after hospital discharge. Rotavirus disease severity (assessed with a 20-point score), duration of rotavirus shedding, and seroresponse to rotavirus were compared between HIV-infected and HIV-uninfected children. FINDINGS: 786 inpatients (median age 8 months, 271 [34%] of whom were HIV-1-infected) and 400 outpatients (median age 9 months, 65 [16%] of whom were HIV-infected) were enrolled. Rotavirus was detected less frequently among HIV-infected children (102 of 336 [30%]) than among HIV-uninfected children (348 of 850 [41%], (relative risk 0.71 [95% CI 0.53-0.87], p=0.0007). There were no differences in rotavirus disease severity for hospitalised children with and without HIV infection, but HIV-infected children were more likely to die during follow-up (11/50 [22%]) than HIV-uninfected children (0/61, p<0.0001). Of 29 HIV-infected and 45 HIV-uninfected children who completed follow-up, six (21%) HIV-infected children shed rotavirus, compared with two (4%) HIV-uninfected children (4.66 [1.01-21.51], p=0.05), but shedding was not associated with diarrhoea. Three-quarters of children exhibited a four-fold rise of serum IgG or IgA to rotavirus, which did not vary by HIV status. INTERPRETATION: Malawian children with concomitant HIV infection resolved acute rotavirus infections. Rotavirus vaccine safety and immunogenicity in HIV-infected infants should now be determined.

Chi-Square Distribution↗
PubMed1990

Antigenic relationships among human rotaviruses as determined by outer capsid protein VP4.

cDNA clones representing the VP4 gene of symptomatic human rotavirus strain KU (VP7 serotype 1) or DS-1 (VP7 serotype 2) or asymptomatic human rotavirus strain 1076 (VP7 serotype 2) were constructed and inserted into a baculovirus expression vector under the control of the polyhedrin promoter. The resulting recombinants expressed the appropriate authentic VP4 rotavirus outer capsid protein. Guinea pigs immunized with these VP4 proteins developed antibodies that neutralized infectivity of the rotavirus from which the immunizing VP4 was derived. These antisera were then used in neutralization tests to define the extent and distribution of VP4 antigenic polymorphism among human rotaviruses. Three distinct serotypes and one subtype of the VP4 outer capsid protein were identified among 17 human rotavirus strains that had previously been assigned to five distinct VP7 serotypes. For the most part, VP4 serotype segregated independently of VP7 serotype. Ten strains of human rotavirus that were associated with symptomatic infection and that exhibited VP7 serotype 1, 3, 4, or 9 specificity, each possessed a VP4 of the same serotype and subtype, designated VP4 serotype 1A. Both symptomatic human rotavirus strains with VP7 serotype 2 specificity were related by neutralization to the VP4 serotype 1A strains and were classified as a subtype of VP4 serotype 1--i.e., serotype 1B--since viruses of serotype 1A appeared to be prime strains. Four human rotavirus strains that were recovered from healthy infants in newborn nurseries in which virus transmission persisted over a long interval, belonged to VP7 serotype 1, 2, 3, or 4, but each strain possessed the same VP4 antigenic specificity that was designated VP4 serotype 2. Finally, a single strain of symptomatic human rotavirus of VP7 serotype 1 specificity possessed a unique VP4 that was provisionally classified as VP4 serotype 3 but this remains to be confirmed because neutralization tests were performed in only one direction. Among the 10 rotavirus strains whose VP4 gene was previously sequenced, there was complete concordance between assignment of VP4 serotype by neutralization and classification according to VP4 amino acid homology. Thus, rotaviruses that exhibited a VP4 amino acid homology of 89% or greater belonged to the same VP4 serotype and subtype as determined by neutralization. Finally, evidence was obtained that the serotype-specific domain is located on the VP8 subunit of VP4.

Animals↗
PubMed1992

Rotavirus-specific helper T cell responses in newborns, infants, children, and adults.

An obstacle to developing a successful rotavirus vaccine has been the inability to consistently correlate the humoral immune response with protection against disease. Transplacental transfer of maternal rotavirus-specific antibodies may obscure the capacity to discriminate an active from a passively acquired humoral immune response in infants. In an attempt to circumvent this problem, an assay was developed to detect rotavirus-specific helper T cells among circulating mononuclear cells. Rotavirus-specific lymphoproliferative responses and rotavirus-specific neutralizing antibody titers in blood were determined in 11 mother/newborn pairs at the time of delivery and in 54 infants, children, and adults ranging in age from 16 days to 40 years. Only 1 of 11 infants tested between 16 days and 6 months of age had detectable rotavirus-specific helper T cell activity whereas 8 of 11 had circulating rotavirus-specific neutralizing antibodies. Acquisition of rotavirus-specific helper T cell activity over the first few years of life correlated with the age at which infants and young children are known to be infected with rotavirus. These findings support the hypothesis that detection of rotavirus-specific lymphoproliferative activity in infants may more accurately determine previous exposure to rotavirus than detection of rotavirus-specific antibodies.

Adolescent↗
PubMed1997

Visualizing geographic and temporal trends in rotavirus activity in the United States, 1991 to 1996. National Respiratory and Enteric Virus Surveillance System Collaborating Laboratories.

BACKGROUND: Rotavirus is the leading cause of severe pediatric gastroenteritis worldwide. A vaccine may soon be licensed for use in the United States to prevent this disease. To characterize US geographic and temporal trends in rotavirus activity, we made contour maps showing the timing of peak rotavirus activity. METHODS: From July, 1991, through June, 1996, 79 laboratories participating in the National Respiratory and Enteric Virus Surveillance System reported on a weekly basis the number of stool specimens that tested positive for rotavirus. The peak weeks in rotavirus detections from each laboratory were mapped using kriging, a modeling technique originally developed for geostatistics. RESULTS: During the 5-year period 118,716 fecal specimens were examined, of which 27,616 (23%) were positive for rotavirus. Timing of rotavirus activity varied by geographic location in a characteristic pattern in which peak activity occurred first in the Southwest from October through December and last in the Northeast in April or May. The Northwest exhibited considerable year-to-year variability (range, December to May) in the timing of peak activity, whereas the temporal pattern in the remainder of the contiguous 48 states was relatively constant. CONCLUSION: Kriging is a useful method for visualizing geographic and temporal trends in rotavirus activity in the United States. This analysis confirmed trends reported in previous years, and it also identified unexpected variability in the timing of peak rotavirus activity in the Northwest. The causes of the seasonal differences in rotavirus activity by region are unknown. Tracking of laboratory detections of rotavirus may provide an effective surveillance tool to assess the impact of a rotavirus vaccination campaign in the United States.

Humans↗
PubMed1999

Reappraisal of the Peruvian and Brazilian lower titer tetravalent rhesus-human reassortant rotavirus vaccine efficacy trials: analysis by severity of diarrhea.

OBJECTIVES AND METHODS: With the purpose of better understanding the efficacy of the lower titer [4 x 10(4) plaque-forming units (pfu)] tetravalent rhesus-human reassortant rotavirus vaccine (RRV-TV) against diarrheal episodes of different severities, the Peruvian and Brazilian efficacy data were reanalyzed with a 20-point scoring system. Mild, moderate/severe and very severe rotavirus diarrhea were scored as 0 to 8, 9 to 14 and >14, respectively. RESULTS: In the Peruvian study one dose of vaccine yielded 64% (P = 0.04) protection against pure cases of rotavirus disease (i.e. those in which no other enteropathogen was found) with clinical scores ranging from 9 to 14. Protective efficacy against very severe rotavirus gastroenteritis could not be assessed because of the small number of cases. In Brazil there was a trend in preventing "all" and "pure" cases of rotavirus diarrhea scored 9 to 14 (44%, P = 0.06, and 45%, P = 0.08, respectively) and the vaccine was 75% (P = 0.02) protective against pure rotavirus diarrhea scored >14. No protection was observed for mild rotavirus diarrhea (scores <9). These data were compared with those from trials in Venezuela (4 x 10(5) pfu/dose), US (4 x 10(4) pfu/dose and 4 x 10(5) pfu/dose) and Finland (4 x 10(5) pfu/dose). Combining the Peruvian (one dose, pure cases) and Brazilian studies together, the levels of protection against 9- to 14-scored rotavirus diarrhea are comparable with those from the Venezuelan (47%) and American (57, 57 and 65%) efficacy trials. In Brazil the level of protection (75%) against pure, >14-scored rotavirus diarrhea is similar to the efficacy rates yielded in the three US trials (82, 80 and 69%) and the Finnish trial (100%) for episodes of the same severity. CONCLUSIONS: Our reanalysis provides evidence that, at least against moderate/severe rotavirus gastroenteritis, RRV-TV, 4 x 10(4) pfu/dose is potentially as efficacious as RRV-TV, 4 x 10(5) pfu/dose, even in settings with very high rotavirus disease burden. The reanalysis of the Peruvian data suggests that one and three vaccine doses may yield similar efficacy rates. It is also suggested that vaccine efficacy against most severe episodes in Peru and Brazil was not evident because of the trial design used in those studies (i.e. prospective, active home surveillance rather than a catchment trial), resulting in too few cases of severe disease even in the placebo group. To confirm these findings, future trials with this vaccine are necessary in developing countries with high diarrhea morbidity rates. These trials should use catchment designs and focus on the evaluation of the efficacy of one or three doses of RRV-TV against moderate to severe/very severe rotavirus diarrhea.

Animals↗
PubMed2004

Natural rotavirus infection is not associated to intussusception in Mexican children.

AIMS: To determine whether natural rotavirus infection or infection by another enteropathogen is associated to intussusception (IS); and to describe the seasonality of IS compared with severe diarrhea (SD) and rotavirus SD in Mexican children. METHODS: A prospective, observational, multicenter and case-control study was conducted in Mexico City from December 1999 to February 2001. Cases were children younger than 1 year old hospitalized for IS; diagnosis was made by clinical features, radiologic and/or surgery findings. Controls were children younger than 1 year old hospitalized for another disease than a gastrointestinal illness (NGI). Cases and controls were paired by age and date of admission (+/-3 months; for both), in a 1:2 ratio. A surveillance of IS cases, SD and rotavirus SD episodes was conducted during the study period. Stool samples collected soon after IS resolution or at admission were tested for rotavirus, adenovirus, astrovirus, bacteria and parasites. RESULTS: Thirty cases of IS and 60 controls with NGI were studied. Rotavirus was not detected in any case of IS. Adenovirus (17%) was the only enteropathogen detected in IS. Rotavirus (8%), adenovirus (2%), astrovirus (2%) and bacteria (2%) were detected in NGI. Rotavirus infection was not associated with IS (odds ratio, 0; 95% confidence interval, 0-2.9), whereas adenovirus infection was strongly associated as risk factor for IS (odds ratio undefined; P = 0.003), compared with NGI. Seasonal variation in admissions for IS was small, whereas admissions for SD and rotavirus SD showed a marked seasonal increase during fall-winter. CONCLUSIONS: In Mexican children, rotavirus infection was not associated to IS; whereas a significant association was observed between adenovirus and IS. Also there was no increase in IS cases during the sharply defined fall-winter rotavirus outbreak. Observations from this controlled study suggest that natural rotavirus infection is not a risk factor for IS. This information may have implications for development of a safer and effective rotavirus vaccine.

Adenoviridae Infections↗
PubMed2007

Rotavirus: to the gut and beyond!

PURPOSE OF REVIEW: Rotavirus causes severe gastroenteritis in children. A principle of rotavirus pathogenesis has been that the infection remains localized to epithelial cells in the small intestine. This dogma was challenged by recent findings of rotavirus in the serum of experimentally infected animals and children with diarrhea. Repeated associations of rotavirus infections with a wide range of nongastroenteric clinical manifestations in humans were considered spurious because of lack of proof that rotavirus escaped the intestine. New data outlined in this review, however, show that rotavirus routinely infects systemically and highlight controversies and future research questions. RECENT FINDINGS: Rotavirus antigens (antigenemia), RNA, or infectious virus (viremia) has been demonstrated in the serum and many extraintestinal tissues in all experimental animal models. Rotavirus antigens and RNA have been detected in the sera of children with rotavirus diarrhea. The tissues and cell types that support rotavirus replication outside the intestine and the consequences of extraintestinal reservoirs of infection are beginning to be examined. SUMMARY: Rotavirus infection is systemic, with an acute active viremia and extraintestinal replication. The impact of systemic rotavirus on disease burden remains to be determined.

Animals↗
PubMed1997

Detection of group B rotaviruses in fecal samples from diarrheic calves and adult cows and characterization of their VP7 genes.

Groups A, B, and C rotaviruses have been identified in cattle. Group B rotaviruses are associated with sporadic cases of diarrhea in calves and adult cows. From diagnostic submissions to our laboratory, 90 fecal samples from cases of calf diarrhea, 81 fecal samples from cases of adult cow diarrhea (winter dysentery), and 20 fecal samples from case control normal adult cows were tested for group B rotaviruses by polyacrylamide gel electrophoresis (PAGE), and reverse transcription (RT)-PCR (targeting 279 bp of the VP7 gene). In addition, 53 fecal samples from diarrheic adult cows were tested for group B rotaviruses by immune electron microscopy (IEM). By RT-PCR, five samples from calves were group B rotavirus positive (5.6%). Fifteen samples from adult cows with diarrhea were group B rotavirus positive (18.5%), and none of the control fecal samples from normal cows were positive for group B rotaviruses. By PAGE, one calf sample (RT-PCR positive) was group B rotavirus positive (short electropherotype), but none of the adult cow samples were positive for group B rotaviruses. By IEM, 5 (9.4%) of the 53 fecal samples from diarrheic adult cows were group B positive (all were also RT-PCR positive). The VP7 genes of three strains (WD653 from an adult cow and the ATI and Mebus calf strains) were sequenced. The VP7 genes from the three bovine strains showed high (over 90%) nucleotide and deduced amino acid homologies, but lower homologies (48 to 61%) were seen between these genes and the genes from rodent (IDIR) and human (ADRV) group B rotaviruses. Although there were some differences of degree, all inoculated gnotobiotic calves (n = 6) showed abnormal feces between 1 and 3 days after inoculation with each of three strains of group B bovine rotaviruses, and group B rotaviruse, were detected in the feces for up to 2 weeks by RT-PCR but for shorter periods by PAGE or IEM.

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