Detection of rotavirus RNA in cerebrospinal fluid in a case of rotavirus gastroenteritis with febrile seizures.
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OBJECTIVE: National estimates of hospitalizations for rotavirus, the leading cause of acute gastroenteritis (AGE) in children, have been used to establish the need for rotavirus vaccines. A previous method directly estimated discharges by using the rotavirus-specific International Classification of Diseases (ICD) code, but this method has not been validated. Our study evaluated the sensitivity of the rotavirus ICD code among children hospitalized for AGE by using active surveillance for rotavirus at a tertiary children's hospital. DESIGN: We compared data for rotavirus-coded hospital discharges in 2000-2001 at Cincinnati Children's Hospital Medical Center with data on laboratory-confirmed cases of rotavirus obtained from active surveillance. We estimated additional rotavirus hospitalizations by extrapolating the proportion of rotavirus-positive results from active-surveillance cases to those with an unknown rotavirus status. RESULTS: Of 767 cases of AGE-related discharge codes, 103 (13%) were coded as rotavirus, 91% (94 of 103) of which were laboratory-confirmed diagnoses. Among all children discharged with an AGE-related illness, 260 (34%) were enrolled in active surveillance, of whom 155 (60%) tested positive for rotavirus. An additional 47 laboratory-confirmed rotavirus-case patients not enrolled in active surveillance yielded a total of 202 rotavirus cases and a maximum sensitivity of the rotavirus code of 47%. Extrapolation indicated that an additional 170 untested children might be rotavirus-positive, yielding a total of 372 rotavirus hospitalizations and a minimum sensitivity of the rotavirus code of 25%. CONCLUSIONS: Measurement of rotavirus-coded hospital discharges alone seems to greatly underestimate the true burden of rotavirus-associated hospitalizations. The numbers of national rotavirus hospitalization discharges may be substantially greater than previously estimated.
BACKGROUND: Rotavirus infection is thought to be confined to the intestine. Reports of rotavirus RNA in the cerebral spinal fluid and serum of children infected with rotavirus suggest the possibility that rotavirus escapes the intestine into the circulatory system. We assessed whether rotavirus antigen, RNA, or both, were present in serum samples from immunocompetent rotavirus-infected children and animals. METHODS: We obtained sera from immunocompetent mice, rats, rabbits, and calves 1-10 days after inoculation with rotavirus or matched vehicle. We obtained sera retrospectively from immunocompetent children diagnosed with rotavirus diarrhoea (n=33), healthy children (n=6) and adults (n=12), children convalescing from rotavirus (n=6), and children with non-rotavirus diarrhoea (n=11). Samples were analysed for the presence of rotavirus antigen or RNA by EIA or RT-PCR, respectively. FINDINGS: Rotavirus antigen was present in sera from rotavirus-infected animals, but not in sera from control animals. Infectious rotavirus or rotavirus RNA was detected in sera of mice and calves, respectively. Antigen was present in 22 of 33 serum samples from children with confirmed rotavirus infection but in none of 35 samples from controls. Detection of serum antigen was inversely related to the number of days between symptom onset and sample collection, and directly related to stool antigen concentration. Rotavirus RNA was detected by RT-PCR in three of six rotavirus-positive sera. INTERPRETATION: Rotavirus can escape the gastrointestinal tract in children, resulting in antigenaemia and possible viraemia. This finding is important for the understanding of the pathogenesis, immunology, and clinical manifestations of rotavirus infection.
In order to evaluate mucosal antibody responses to rotavirus, an enzyme-linked immunospot (ELISPOT) assay was adapted to enumerate antibody-secreting cells (ASC) in the mesenteric lymph nodes (MLN), lamina propria (LP) of the small intestine and spleens of gnotobiotic pigs orally inoculated with porcine rotaviruses (SB1A and Gottfried). Rotavirus-specific IgM ASC occurred by post-inoculation day (PID) 3, and numbers peaked in spleen and MLN tissues by PID 7 and in intestinal LP by PID 7-14. Numbers of rotavirus specific IgA and IgG ASC in these tissues peaked at PID 14-21. Rotavirus specific IgA ASC were predominant in the gut and IgA to IgG rotavirus specific ASC ratios were highest for all rotavirus antigen coatings in the gut LP. However, the relative ratios of specific IgA to IgG ASC were lower (ratios of 5 to 7) against combined structural and nonstructural viral antigens (rotavirus-infected fixed cell ELISPOT plates) than ratios (13 to 46) against only viral structural antigens (rotavirus-coated ELISPOT plates), indicating that there were proportionately more specific IgG ASC to the nonstructural viral antigens in the LP, the tissue adjacent to the site of rotavirus replication in the intestine. In the node cells (spleen and MLN) rotavirus-specific IgA to IgG ASC ratios were lowest and against the various ELISPOT rotavirus coatings ranged from 0.7 to 4. Gnotobiotic piglets inoculated at different ages with porcine rotaviruses generally showed similar specific immunoglobulin (Ig) ASC responses to rotavirus infection, along with similar diarrhea and virus shedding patterns in the different age groups. However, the numbers of specific IgA ASC in the MLN of 3-4 week old pigs were higher than those of 3-5 day old pigs. Although challenge of SB1A or Gottfried rotavirus-inoculated pigs with SB1A (G4P7) or Gottfried (G4P6) rotavirus revealed a high degree of protection from diarrhea and virus shedding, greater numbers of specific IgM ASC were observed in spleen after challenge of SB1A-inoculated pigs with Gottfried rotavirus (same G type, distinct P type). Thus, by using the ELISPOT technique, we successfully measured intestinal mucosal antibody-related responses to rotavirus in gnotobiotic pigs. Moreover, our results support the use of gnotobiotic piglets as an animal model to evaluate active antibody responses and protection against rotavirus infection and disease.
Little is known about the epidemiology of rotavirus infection in Turkey. The aim of the study was to determine the incidence and clinical significance of rotavirus gastroenteritis, in view of the potentially available prevention by rotavirus vaccination. The study also sought to determine possible risk factors for rotavirus gastroenteritis. Therefore, 920 children under five years of age with acute gastroenteritis admitted to three pediatric hospitals in Izmir were studied. Rotavirus was identified in 39.8% of the children. Most children with rotavirus gastroenteritis (80.7%) were younger than two years of age. Marked seasonality of rotavirus gastroenteritis was observed, with a peak incidence from January to March. A total of 91% of rotavirus strains that were typed were of serotypes G 1-4. There was no significant difference among rotavirus-positive and rotavirus-negative patients with regard to family income. Compared with children who were exclusively breast-fed, those who were not exclusively breast-fed were at a two-fold greater risk of rotavirus diarrhea. Rotavirus gastroenteritis was significantly more severe than non-rotavirus gastroenteritis; 69% of children with rotavirus infection had severe gastroenteritis (score > or = 11). In conclusion, rotavirus is the most common cause of severe gastroenteritis among children under five years of age in Izmir. A new potent rotavirus vaccine, when available, will provide effective protection against severe rotavirus infection. Promotion of breast-feeding would augment the impact of rotavirus vaccines in preventing severe childhood diarrhea.
To compare epidemiologic features and genetic characteristics of group A rotaviruses causing diarrhea in children and adults, a survey was conducted in Wuhan, China, during the period of Dec. 2000-May 2006. A total of 3839 stool specimens from diarrheal patients from eight hospitals were analyzed. Winter seasonality was observed for rotavirus diarrhea in both adults and children, showing overall rotavirus-positive rates of 9.0 and 23.9%, respectively. Throughout the study period, G3 was the most frequent G serotype in both adults and children (detection rates 86.2 and 87.8%, respectively), and was mostly associated with VP4 genotype P[8], VP 6 genotype II (subgroup II), and NSP4 genotype B. G3 rotaviruses were differentiated into eight electropherotypes, among which seven types were found in specimens from both adults and children. VP7 gene sequences of G3 rotaviruses from adults and children (6 and 4 strains, respectively), detected in different years and different hospitals, showed extremely high sequence identities (99-100%) to each other and to a few G3 rotavirus strains reported in Asia. However, lower sequence identities (82-96%) were observed to most of the human and animal G3 rotaviruses reported so far, including some Chinese strains. These findings indicate that in Wuhan, China, epidemic and genetic features of rotaviruses are similar in adults and children, and it has been suggested that G3 rotaviruses that might have originated from the same rotavirus were circulating among children and adults as prevailing viruses. In this study, two rotavirus strains, G9P[8] strain L169, derived from an adult, and G4P[6] strain R479, derived from a child, were isolated and genetically analyzed. The VP7 gene of L169 belongs to a major lineage of G9 rotaviruses that are globally widespread, but is distinct from G9 rotaviruses reported previously in China. The strain R479 had a VP7 gene which was divergent from most G4 human rotaviruses and showed an unusual dual subgroup specificity, I + II. The R479 VP6 gene does not belong to the main clusters of subgroup I and II rotaviruses phylogenetically, but is related to those of the porcine rotaviruses and some unusual human rotaviruses represented by the RMC321 strain isolated in eastern India.
BACKGROUND: Human group B rotavirus was first identified as causative agent of a large outbreak of severe gastroenteritis affecting more than 1 million people, predominantly adults in China in 1982-1983. In spite of serological evidences for the presence of group B rotavirus in many countries of the world, the virus has been detected only from China, India and Bangladesh, where most of the cases were from adults. OBJECTIVES: To ascertain the role of group B rotavirus as an aetiological agent of diarrhoea among children in Kolkata, India. STUDY DESIGN: An active surveillance was conducted for rotavirus infection in children in a leading referral paediatric hospital and a few samples were also collected from adults of another hospital in Kolkata, India over a period of 3 years (2002-2004). After primary screening of rotaviruses by RNA electrophoresis in polyacrylamide gel, 200 of 412 samples negative by PAGE were screened by reverse transcription polymerase chain reaction for group B rotaviruses. The group B rotavirus positives samples were also confirmed by dot-blot hybridization. RESULT: During the study period, we detected 37 (18.5%) sporadic cases of human group B rotavirus infection in children below 3 years of age of which 15 (7.5%) showed mixed infection with group A rotaviruses by RT-PCR. In dot-blot hybridization studies the RNA of all rotavirus positive samples hybridized with the nonisotopic psoralen-biotin labeled total RNA probe generated from a human group B rotavirus CAL-1 strain confirming the samples as group B rotaviruses. CONCLUSION: The shift in age preference of group B rotavirus infection from adult to children and mixed infection of group B and group A rotaviruses reveals the importance of group B rotavirus as an etiological agent of childhood diarrhoea. Therefore, future vaccination strategy should include both group A and B rotaviruses to control rotavirus diarrhoea.