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PubMed2005

Peak rotavirus activity shifted from winter to early spring in Japan.

BACKGROUND: Since 1910, there have been many studies on acute gastroenteritis in children in Japan. These diseases, namely Kasei-shoni-kolera (pseudocholera infantum) or banshu-otosho (late autumn vomiting disease), are historically known to occur in the cooler season with a peak in November or December. Earlier we confirmed their causation by rotaviruses but found peaks in January or February from 1974 to 1981. The aim of the present study was to confirm the temporal shift in peak rotavirus activity. METHODS: Under the National Epidemiological Surveillance of Infectious Diseases program from 1983 through 2003, rotavirus positive patients 0-3 years old and clinically diagnosed with "infantile vomiting and diarrhea" at sentinel clinics were examined. Fecal samples were screened by electron microscopy and/or using commercial latex agglutination kits at prefectural/municipal Public Health Institutes, and we determined the trend for the "peak" month during 21 seasons. RESULTS: Peak rotavirus activity shifted gradually from January to March during the 21 consecutive seasons. The mean duration from December to the peak month (mean beginning peak duration) of the rotavirus season significantly varied among 3 periods of 7 consecutive seasons (1.7 +/- 0.5 months in 1982/1983-1988/1989, 2.3+/-0.8 months in 1989/1980-1995/1996, and 3.1 +/- 0.7 months in 1996/1997-2002/2003, respectively; P = 0.0026 by 1-way analysis of variance). This time series shift in the peak rotavirus infection was statistically significant (P = 0.0003 for trend). CONCLUSION: Our findings confirmed that the temporal trend in peak rotavirus activity in Japan has shifted gradually from winter to early spring for unknown reasons.

Age Distribution↗
PubMed2005

Evaluation of safety, immunogenicity and efficacy of an attenuated rotavirus vaccine, RIX4414: A randomized, placebo-controlled trial in Latin American infants.

BACKGROUND: A live attenuated monovalent rotavirus vaccine RIX4414 was developed with a human strain of G1P1A P[8] specificity to reduce the rotavirus burden in children. METHODS: A double blind, randomized, placebo-controlled study evaluated the efficacy, immunogenicity, safety and reactogenicity of 2 oral doses of RIX4414 (10(4.7), 10(5.2) or 10(5.8) focus-forming units) at 2 and 4 months coadministered with routine vaccinations and oral poliovirus vaccine given for study purposes at least 14 days apart. The 2155 infants (1618 vaccine/537 placebo) enrolled in Brazil, Mexico and Venezuela were followed until 1 year of age. RESULTS: Antirotavirus IgA seroconversion rates 2 months after dose 2 ranged between 61% (10(4.7) ffu group) and 65% (10(5.8) ffu group), and most of the infants had seroprotective levels of antibodies to coadministered routine vaccinations. The reactogenicity profile of RIX4414 was similar to that of the placebo, and no vaccination-related serious adverse events were reported. Protective efficacy against severe and any rotavirus gastroenteritis from 15 days post-dose 2 was highest in the 10(5.8) ffu group [86%; 95% confidence interval (95% CI), 63-96% and 70% (95% CI 46-84%), P < 0.001, 2-sided Fisher's exact test]. The efficacy against hospitalization was 79% (95% CI 48-92%) for pooled vaccine groups. Multiple rotavirus serotypes [G1 (50%), G9 (40%), G2, G3 and G4] were identified from gastroenteritis stools (enzyme-linked immunosorbent assay and reverse transcription-polymerase chain reaction) during the study period. For severe gastroenteritis caused by G9 serotypes, the protection reached 77% (95% CI 18-96%) in the 10(5.8) ffu group, providing proof of concept that the monovalent G1P1A P[8] human rotavirus vaccine elicits cross-protection against the G9 strain. A reduction in any and severe rotavirus gastroenteritis was already observed at post-dose 1 (period: day of dose 1 to 14 days post-dose 2) in vaccinees compared with placebo recipients. CONCLUSIONS: Two doses of RIX4414 are highly efficacious, providing cross-protection (G1 and G9 strains, prevalent during this study) and early protection against any and severe rotavirus gastroenteritis and hospitalization to infants in Latin America.

Administration, Oral↗
PubMed1987

Avirulent rotavirus infections protect calves from disease with and without inducing high levels of neutralizing antibody.

Two bovine rotaviruses, C3-160 and 17/4, which multiplied in calves without inducing disease, were studied for their ability to protect against disease caused by a virulent rotavirus, CP-1. Rotavirus 17/4 and the virulent virus CP-1 cross-neutralized poorly and, on the basis of 20-fold differences in neutralization titres, belonged to different serotypes. Rotaviruses C3-160 and CP-1 were more closely related: neutralization of the CP-1 virus by C3-160 antisera was within 20-fold of the homologous titre although neutralization of the C3-160 virus by CP-1 antisera was not. Nine gnotobiotic calves infected with either C3-160 or 17/4, had rotavirus antibody in their faeces and/or serum 21 days after oral inoculation as detected by indirect immunofluorescence and IgG, IgM and/or IgA antibodies by ELISA. As expected from the antigenic relationships between the viruses, the sera and faeces from the four calves infected with C3-160 contained moderate levels of neutralizing antibody to the virulent virus CP-1 and the sera and most of the faeces from the five calves infected with 17/4 contained undetectable or low levels. When challenged with CP-1 on day 21, four age-matched controls developed disease whereas all of four calves previously infected with C3-160 and four of five calves previously infected with 17/4 were protected from disease. It was concluded that avirulent rotavirus infection provided protection against disease caused by a virulent rotavirus even when one of the avirulent viruses was poorly related to the virulent virus by neutralization. Mechanisms other than neutralizing antibody appeared to be important in protection.

Animals↗
PubMed1999

Burden of human rotavirus-associated hospitalizations in three geographic regions of Hungary.

Data on hospital admissions and laboratory reports were used to estimate the number of hospitalizations of children aged 14 y or less in three geographic regions of Hungary due to group A rotavirus infection. Between January 1993 and December 1996, 9182 hospitalizations for gastroenteritis occurred, of which 1946 (21%) were associated with rotavirus infection. Most (90%) of the rotavirus detections were among children aged 4 y or less. By extrapolation, an estimated 5000 rotavirus-related hospitalizations (8.4/1000 children aged 4 y or less/y) occurred in Hungary during the study period. Marked seasonality of rotavirus infections was observed, with a peak of incidence from December to February. Rotaviruses with "long" RNA electropherotypes predominated each year, but in 1995/1996 20% of electropherotypes in the Budapest area were "short". Effective surveillance is required for all children hospitalized for diarrhoea as part of a rotavirus immunization program in Hungary.

Child, Preschool↗
PubMed1991

Rotavirus excretion by village pigs in Papua New Guinea.

Cohort studies were conducted on 29 pigs from 3 villages in the Highlands of Papua New Guinea. Animals ranged in age from 9 d to 5 m old. Three hundred and twenty nine faecal samples were collected from individual pigs followed over 3 to 6 w periods, and were examined for group A rotavirus antigen by ELISA, and rotaviral genomic RNA by polyacrylamide gel electrophoresis (PAGE). Electron microscopy was also conducted on selected samples. Group A rotavirus was detected in the faeces of 16 pigs with infected individuals coming from all villages. Non-group A rotavirus resembling group C was found in faeces from pigs from 2 villages. All of the group A rotaviruses examined had the same electrophoretype and this was distinct from that of the common type infecting humans in the area at the time of the study. None of the group A positive samples reacted with monoclonal antisera specific for human group A rotaviruses of serotypes 1, 2, 3, 4, or 8. The non-group A rotaviruses also all had identical electrophoretypes. In contrast to previous findings in intensive piggeries, rotavirus infection did not occur in all young pigs and was not limited to young animals under 2 m of age. Infected pigs varied in age from 12 days to 20 weeks of age. This pattern of infection was attributed to the non-intensive husbandry situations in the villages, with less opportunity for transmission to occur than in intensive piggeries.

Animal Husbandry↗
PubMed1999

[Trial of oral administration of Bifidobacterium breve for the prevention of rotavirus infections].

It was investigated that Bifidobacterium breve YIT4064 (B. breve YIT4064), Which had augmented IgA production and prevented rotavirus-induced diarrhea in mice, prevented rotavirus infection in infants. The effect of B. breve YIT4064 was evaluated in ten infants from an infants home who received 50 mg of the bacterium every day for 28 days (the B. breve group). Nine infants did not receive this (the control group). Though rotavirus shedding in the control group was detected from 2 (a total of 5 stool samples) of 9 infants (a total of 112 stool samples), it was not detected in any infants (a total of 133 stool samples) in the B. breve group during the administration period. From day 8 to day 14 of the test, rotavirus shedding was detected from 4 of 32 stool samples in the control group, but was not detected at all from 38 stool samples in the B. breve group. The frequency of rotavirus shedding in the B. breve group was significantly lower than that in the control group. Further, the frequency in appearances of anti-IgA in stool samples in the B. breve group showed a tendency to increase in comparison with the control group from day 8 to 14 of the test. The oral administration of B. breve YIT4064 significantly decreased rotavirus shedding in stool samples and prevented rotavirus infection.

Administration, Oral↗
PubMed2006

Real-time reverse transcription-PCR for detection of rotavirus and adenovirus as causative agents of acute viral gastroenteritis in children.

Viral pathogens are the most common cause of gastroenteritis in developed countries. Human rotavirus and adenovirus infections are major causes of acute outbreaks and sporadic cases of gastroenteritis, occurring primarily among children less than 2 years of age. Patient hospitalization is often required, with enormous infection control implications. This work describes the development of real-time PCR assays for the detection of group F adenovirus, rotavirus A, and rotavirus C from stool specimens. Two hundred twenty stool samples from pediatric patients exhibiting symptoms of diarrhea and/or vomiting were examined. PCR results were compared with those of virus detection by electron microscopy and latex agglutination antigen detection. The incorporation of an internal-control RNA that was spiked into individual stool extracts functioned as an internal validation for the reporting of PCR-negative results. Rotavirus C was not detected by real-time PCR in the patient stool samples examined. Real-time reverse transcription-PCR resulted in 175% and 111% increases in the rates of detection of adenovirus F and rotavirus A, respectively, compared with latex agglutination testing. Molecular detection increased the number of stool specimens in which causative agents of gastroenteritis were identified by 155% compared to electron microscopy. Genotyping of a proportion of the rotavirus and adenovirus strains identified only genotype G1 rotavirus and both adenovirus genotypes 40 and 41 in circulation within the patient cohort examined. The results highlight the significance of rapid molecular methods for the routine screening of stool samples in hospital laboratories to provide rapid definitive diagnoses.

Acute Disease↗
PubMed1983

Antigenic relationships among some bovine rotaviruses: serum neutralization and cross-protection in gnotobiotic calves.

A method was further developed to screen non-tissue-culture-adapted bovine rotaviruses for serotype, using a neutralization test with infectious fecal rotavirus. One of those rotaviruses (B223) which was not blocked by antiserum to the neonatal calf diarrhea virus (NCDV) serotype was then adapted to cell culture in the presence of the antiserum for two or more passages and hyperimmune antiserum to this isolate had a 60-fold-higher homologous neutralization titer than with the NCDV serotype rotavirus. Seventy-three isolates were serotyped and eight (11%) were not of the NCDV serotype (bovine rotavirus serotype I). Of these eight, five belonged to the new bovine rotavirus serotype II and three were not typed, indicating the existence of one or more further serotypes. Cross-protection studies in gnotobiotic calves showed that cross-protection only occurred between rotaviruses of the same serotype, and even a minor serotype difference was sufficient for the calves to show a lack of cross-protection. The serotypes (I and II and the three untyped isolates) also showed differences in the rate of migration in polyacrylamide gel electrophoresis of some of their RNA segments (no. 4, 6, 7, 8, 9, 10), indicating that they were of different electropherotypes.

Animals↗
PubMed1986

Clinical and epidemiological features of acute infantile gastroenteritis associated with human rotavirus subgroups 1 and 2.

During a prospective 1-year study rotavirus isolates from 169 children with gastroenteritis were investigated by polyacrylamide gel electrophoresis. A total of 118 (70%) of the strains analyzed contained sufficient viral nucleic acid to give visible electrophoretic patterns; 36% were identified as strains belonging to subgroup 1 (short patterns), and 64% were identified as strains belonging to subgroup 2 (long patterns). The two subgroups cocirculated at equal frequencies during the first 7 months of the year, after which subgroup 1 rotavirus completely disappeared. Subgroup 2 rotavirus occurred throughout the year. No significant differences between the subgroups in relation to age or sex distribution were observed. Fever and temperatures exceeding 39 degrees C were significantly more frequent in children who shed rotavirus subgroup 1. Diarrhea and vomiting occurred at similar rates in both groups of patients, but were more pronounced in children who shed rotavirus subgroup 2. One of three dominant electropherotypic variants of subgroup 2 rotavirus was found to be associated with more intense symptoms, higher rates of hospitalization, and a significantly higher frequency of respiratory symptoms; the clinical picture may indicate that this rotavirus electropherotype has higher virulence.

Adolescent↗
PubMed1988

Development of neutralizing antibodies and group A common antibodies against natural infections with human rotavirus.

We determined the levels of group A common and neutralizing antibodies against human rotavirus in paired serum specimens obtained from 38 infants within 12 days of the onset of diarrhea. Thirty of the infants excreted rotavirus in stools, and eight did not. Nine patients (30%) with rotavirus diarrhea and seven patients (88%) with diarrhea due to other causes had detectable levels (greater than or equal to 1: 80) of immunoglobulin (IgG) common antibodies in acute-phase sera. All the patients with rotavirus diarrhea showed at least fourfold rises in titers of IgG or IgM common antibodies or both, while only two control patients showed significant rises in either IgG or IgM common antibodies in their convalescent-phase sera. Of the 19 patients excreting "short" electropherotypes of rotavirus, 18 showed at least fourfold rises in titers of neutralizing antibodies against serotype 2 human rotavirus but not against serotype 1, 3, or 4. Nine of the ten patients excreting "long" electropherotypes showed significant rises in neutralizing antibodies against serotype 3, and the other patient showed a significant rise in neutralizing antibodies against serotype 1. One patient excreted long and short electropherotypes simultaneously, and he also showed a significant rise in neutralizing antibodies against serotype 2 and 3 viruses. The control patients with diarrhea did not show significant changes in titers of antibodies against any of the serotypes. These results demonstrated that the neutralizing antibody response within 2 weeks after clinical onset is specific for the infecting serotype of rotavirus.

Antibodies, Viral↗
PubMed1990

Choice of reference assay for the detection of rotavirus in fecal specimens: electron microscopy versus enzyme immunoassay.

Two previously demonstrated sensitive and specific enzyme immunoassays (EIAs) for rotavirus, one using polyclonal and monoclonal antisera (TestPack Rotavirus [TPK]; Abbott Laboratories) and the other using only monoclonal anti-rotavirus antibodies (Rotaclone [RTC]; Cambridge BioScience Corporation), were evaluated as potential reference assays for rotavirus testing in comparison with direct negative-staining electron microscopy (EM), the current laboratory standard. Two hundred and seven stool samples collected consecutively during the winter of 1989 from children with acute diarrhea admitted to a ward for infants from 0 to 2 years of age were tested by the EIAs and by EM. TPK specimens were read visually; RTC results were read spectrophotometrically. Specimens with discordant EIA and EM results were further evaluated by a fluorescent focus assay. Specimens positive by EM and those negative by EM but positive by fluorescent focus assay were considered to be positive for rotavirus. Of the 207 stools tested, 35 (17%) were positive for rotavirus by these criteria. EM had a sensitivity of only 80%. Specificities were 100% for RTC and EM and 89% for TPK. These findings indicate that EM, although very specific, is relatively insensitive compared with a highly sensitive monoclonal antibody-based EIA. An EIA with high sensitivity and specificity, such as RTC, is a more appropriate reference standard for rotavirus testing.

Acute Disease↗
PubMed1991

Isolation, characterization, and serial propagation of a bovine group C rotavirus in a monkey kidney cell line (MA104).

A virus (designated the Shintoku strain) which was morphologically indistinguishable from group A rotaviruses was detected in the feces of adult cows with diarrhea in Japan. The virus contained 11 segments of double-stranded RNA and had an electrophoretic migration pattern in polyacrylamide gels similar to that of other group C rotaviruses (4-3-2-2). Feces containing the bovine virus reacted with antiserum to porcine group C rotavirus (Cowden strain) but not group A or B rotaviruses in immunoelectron microscopy. The virus was adapted to serial propagation in roller tube cultures of a rhesus monkey kidney cell line (MA104) by using high concentrations of trypsin. Evidence for viral replication in MA104 cell cultures was demonstrated by immunoelectron microscopy and indirect immunofluorescence by using antiserum to porcine group C rotavirus and by electrophoretic analysis of extracted viral double-stranded RNA. A significant antibody response against the isolate was detected in convalescent-phase sera of cows which excreted the virus: no increased antibody response to bovine group A rotavirus was observed. To our knowledge, this is the first isolation of a group C rotavirus from cattle.

Animals↗
PubMed1992

Isolation of an avianlike group A rotavirus from a calf with diarrhea.

An atypical group A rotavirus (993/83) was isolated from a 3-day-old German calf with diarrhea. It differed from 35 conventional German bovine rotavirus isolates analyzed previously with respect to subgroup (strain 993/83 was non-subgroup I and non-subgroup II), serotype (strain 993/83 showed a two-way cross-reaction with serotype 7 and a one-way cross-reaction with serotype 3), and electropherotype (strain 993/83 showed comigrating gene segments 10 and 11). Isolate 993/83 reacted with only one of four monoclonal antibodies that recognized a common VP6 epitope(s). In addition, VP6 and VP2 of isolate 993/83 showed one-dimensional peptide maps that differed substantially from the peptide maps of VP6 and VP2 from all bovine rotavirus isolates. By RNA-RNA hybridization, the 993/83 probe failed to react with a panel of mammalian rotavirus strains, including bovine rotaviruses. It hybridized, however, to genomic RNA of an avian rotavirus strain. Isolate 993/83 could thus represent a candidate for a natural interspecies transmission of rotavirus between different classes of vertebrates.

Animals↗
PubMed1992

Rotavirus diarrhea in Bangladeshi children: correlation of disease severity with serotypes.

To improve the understanding of the relative importance of serotypes of rotavirus in dehydrating diarrhea, we examined the correlation of clinical characteristics and disease severity with serotype in 2,441 diarrheal episodes among children younger than 2 years of age in rural Bangladesh. Of 764 rotavirus-associated episodes, a single G type (serotype 1, 2, 3, or 4) was determined by oligonucleotide probe in 485 (63%), while 233 episodes were nontypeable. Episodes with G types 2 and 3 were associated with more-severe dehydration than episodes associated with G type 1 or 4 or with nontypeable rotavirus. Episodes did not differ by G type in prevalence of vomiting, copious diarrhea, fever, abdominal pain, or length of treatment center stay. Rotavirus reinfections were detected in seven children, with homologous reinfection (G type 2) in one. Twelve children with diarrhea who died had rotavirus detected in stool specimens within 30 days of death. Children who died were more likely to be malnourished than were surviving children with rotavirus diarrhea. Of 40 specimens tested by polymerase chain reaction, 29 (72.5%) were P type 1, 9 (22.5%) were P type 2, 1 (2.5%) was P type 3, and 1 (2.5%) was nontypeable. One severely symptomatic diarrheal episode was associated with P type 3 rotavirus, a serotype usually found in asymptomatic nursery infections. Although G types 2 and 3 were associated with more-severe dehydration than other serotypes, the differences do not appear to be of major clinical importance. Effective vaccines should protect against all four major G types.

Bangladesh↗
PubMed2001

Epidemiological patterns of rotaviruses causing severe gastroenteritis in young children throughout Australia from 1993 to 1996.

Rotavirus strains that caused severe diarrhea in 4,634 (2,533 male) children aged less than 5 years and admitted to major hospitals in eight centers throughout Australia from 1993 to 1996 were subject to antigenic and genetic analyses. The G serotypes of rotaviruses were identified in 81.9% (3,793 of 4,634) children. They included 67.8% (from 3,143 children) serotype G1 isolates (containing 46 electropherotypes), 11.5% (from 531 children) serotype G2 isolates (27 electropherotypes), 0.8% (from 39 children) serotype G3 isolates (8 electropherotypes), and 1.6% (from 76 children) serotype G4 isolates (9 electropherotypes). G6 (two strains) and G8 (two strains) isolates were identified during the same period. G1 serotypes were predominant in all centers, with intermittent epidemics of G2 serotypes and sporadic detection of G3 and G4 strains. With the exception of two strains (typed as G1P2A[6] and G2P2A[6]) all serotype G1, G3, and G4 strains were P1A[8] and all serotype G2 strains were P1B[4]. Two contrasting epidemiological patterns were identified. In all temperate climates rotavirus incidence peaked during the colder months. The genetic complexity of strains (as judged by electropherotype) was greatest in centers with large populations. Identical electropherotypes appeared each winter in more than one center, apparently indicating the spread of some strains both from west to east and from east to west. Centers caring for children in small aboriginal communities showed unpredictable rotavirus peaks unrelated to climate, with widespread dissemination of a few rotavirus strains over distances of more than 1,000 km. Data from continued comprehensive etiological studies of genetic and antigenic variations in rotaviruses that cause severe disease in young children will serve as baseline data for the study of the effect of vaccination on the incidence of severe rotavirus disease and on the emergence of new strains.

Australia↗
PubMed2004

Diversity of group A human rotavirus types circulating over a 4-year period in Madrid, Spain.

The incidence and distribution of human rotavirus G types among children under 5 years old with acute gastroenteritis were determined over a 4-year period (1998 to 2002) by using monoclonal antibodies and reverse transcription-PCR methods. Rotavirus was detected in 1,155 (31%) of 3,760 specimens tested. Rotavirus was studied in every month of the 48-month survey period. Rotavirus activity occurred mainly (51%) in the typically cooler months in Spain (November to February). The age distribution of rotavirus-positive cases showed that 90% of patients (1,038 of 1,155) were under 2 years old. Rotavirus types were determined for 576 of 1,155 patients (50%). G1 was the main genotype detected (53%), and the second most common was G4 (24%). The G2, G9, and G3 rotavirus types were detected in 14, 6, and 2% of the cases, respectively. Dual infections were detected in only 0.6%. The seasonal distribution of genotypes showed a significant genotypic shift: whereas G4 strains predominated (57%) during the 1998 to 2000 seasons, the G1 gradually increased to account for 75% in the 2000 to 2002 seasons. In addition, the present study reports the first detection of the G9 genotype in human fecal samples in Spain. Therefore, additional types may be required for vaccine development strategies that currently target only types G1 to G4.

Age Distribution↗
PubMed2006

Rotavirus activates JNK and p38 signaling pathways in intestinal cells, leading to AP-1-driven transcriptional responses and enhanced virus replication.

Rotavirus infection is known to regulate transcriptional changes in many cellular genes. The transcription factors NF-kappaB and AP-1 are activated by rotavirus infection, but the upstream processes leading to these events are largely unidentified. We therefore studied the activation state during rotavirus infection of c-Jun NH2-terminal kinase (JNK) and p38, which are kinases known to activate AP-1. As assessed by Western blotting using phospho-specific antibodies, infection with rhesus rotavirus (RRV) or exposure to UV-psoralen-inactivated RRV (I-RRV) resulted in the activation of JNK in HT-29, Caco-2, and MA104 cells. Activation of p38 during RRV infection was observed in Caco-2 and MA104 cells but not in HT-29 cells, whereas exposure to I-RRV did not lead to p38 activation in these cell lines. Rotavirus strains SA11, CRW-8, Wa, and UK also activated JNK and p38. Consistent with the activation of JNK, a corresponding increase in the phosphorylation of the AP-1 component c-Jun was shown. The interleukin-8 (IL-8) and c-jun promoters contain AP-1 binding sequences, and these genes have been shown previously to be transcriptionally up-regulated during rotavirus infection. Using specific inhibitors of JNK (SP600125) and p38 (SB203580) and real-time PCR, we showed that maximal RRV-induced IL-8 and c-jun transcription required JNK and p38 activity. This highlights the importance of JNK and p38 in RRV-induced, AP-1-driven gene expression. Significantly, inhibition of p38 or JNK in Caco-2 cells reduced RRV growth but not viral structural antigen expression, demonstrating the potential importance of JNK and p38 activation for optimal rotavirus replication.

Animals↗
PubMed1985

Diagnosis of rotavirus infection with cloned cDNA copies of viral genome segments.

The diagnostic potential of cloned cDNA copies of human rotavirus (strain WA) genome segments for the detection of rotavirus in clinical specimens has been determined. A hybridization assay in which a mixture of 32P-labeled cDNAs representing the 11 rotavirus segments was used as a probe compared favorably with three frequently used diagnostic tests for rotavirus in terms of both specificity and sensitivity. Significantly, clinical isolates could be readily distinguished when cloned cDNA copies of individual genome segments were used independently as a probe. In assays in which genome RNA from rotaviruses of known subgroups and serotypes were tested, cloned probes that encode nonstructural viral proteins hybridized efficiently to genome RNAs of all strains, whereas cloned probes corresponding to genome segments 6 and 9 exhibited the potential for differentiating strains of different subgroups and serotypes. Cloned cDNA copies of rotavirus genome segments therefore offer considerable potential for improved general diagnosis of rotavirus in clinical specimens, as well as for epidemiological studies in which virus isolates can be distinguished on the basis of nucleotide sequence homology of individual genome segments.

Cloning, Molecular↗