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S Urasawa

Publications and source records attributed to S Urasawa.

At least 91 records · Page 5Linked to original sources
PubMed1990

Serotype analysis of group A human rotavirus related to acute gastroenteritis in winter in Gifu city.

A total of 348 fecal specimens collected from children with acute gastroenteritis in Gifu city over three consecutive winter seasons (from November 1986 to March 1989) were examined for group A human rotavirus (HRV) by either a commercial test kit or a sandwich enzyme-linked immunosorbent assay (sandwich ELISA). One hundred twelve of the 173 group A HRV-positive specimens were further subjected to serotype determination by ELISA with four serotype-specific monoclonal antibodies to VP7 (ELISA-serotyping). Ninety-one specimens (81.3%) were successfully serotyped: 41 (36.6%) were serotype 1, 13 (11.6%) serotype 2, 27 (24.1%) serotype 3, and 10 (8.9%) serotype 4. The serotypes of the remaining 21 (18.8%) could not be determined. The predominant serotype of HRV that prevailed in Gifu city changed every winter: serotype 3 (63.4%) was most prevalent in the 1st winter, serotype 4 (42.9%) in the 2nd winter, and serotype 1 (64%) in the 3rd winter.

Adolescent↗
PubMed1990

Group C rotavirus infection and infiltration.

We recently found many cases of gastroenteritis involving older children and sometimes other members of their families. We suspected epidemiological infiltration of an atypical rotavirus (RV). PAGE analysis of 130 cases in 1985 and 1986 revealed that the RNA profiles were similar to that of group C RV in 12 cases. We performed IEM analysis on one of the strains and found it to be group C. The group C RV was detected for 2 months toward the end of the outbreak of group A RV infection. The ages of the group C RV infected patients were 4 years or older. We also studied the prevalence of antibody to group C in the stored sera that had been collected since 1971. Antibody was seldom found in the sera of patients under 3 years of age but it was found in 20-40% of those who were 4 years or older. The clinical manifestations of the group C RV infection were similar to those of group A, except that body temperature was slightly lower and other symptoms were generally milder.

Antibodies, Viral↗
PubMed1990

Antigenic and genetic analyses of human rotavirus with dual subgroup specificity.

In our previous study (S. Urasawa, T. Urasawa, K. Taniguchi, F. Wakasugi, N. Kobayashi, S. Chiba, N. Sakurada, S. Morita, O. Morita, M. Tokieda, T. Kawamoto, K. Minekawa, and M. Oseto, J. Infect. Dis. 160:44-51, 1989) of antigenic characterization of about 300 human rotavirus (HRV) isolates collected at different localities in Japan, we found 4 HRV isolates having unique antigenic and genetic constructions. The four strains possessed both subgroup I and subgroup II antigens, serotype 3 antigen, and a long RNA electropherotype. The reactivity pattern of these four HRV isolates with three monoclonal antibodies (MAbs) directed to an outer capsid protein, VP4, and with one MAb directed to an inner capsid protein, VP2, was clearly different from those of usual subgroup II HRVs having serotype 1, serotype 3, or serotype 4 specificity and a long RNA pattern, whereas their reactivity pattern was similar to that of strain K8 (subgroup II, serotype 1), which possessed unique VP4 and VP2 proteins. RNA-RNA cross-hybridization analysis indicated that while the four isolates were genetically distinct from the two genetic groups of HRV reported previously, i.e., the Wa family (strains KU, S3, and YO) and the DS-1 family (strain S2), they were closely related to strain K8, a strain having unique antigenic and genetic properties (K. Taniguchi, K. Nishikawa, T. Urasawa, S. Urasawa, K. Midthun, A. Z. Kapikian, and M. Gorziglia, J. Virol. 63:4101-4106, 1989).

Antigens, Viral↗
PubMed1990

Porcine rotaviruses antigenically related to human rotavirus serotypes 1 and 2.

Fecal samples from rotavirus-infected piglets were characterized by a serotyping enzyme-linked immunosorbent assay (ELISA) by using monoclonal antibodies (MAbs) specific to human serotypes 1, 2, 3, and 4 (D. O. Matson, M. K. Estes, J. W. Burns, H. B. Greenberg, K. Taniguchi, and S. Urasawa, submitted for publication). Rotavirus in 19 of 25 specimens tested from two herds of pigs from Buenos Aires province, Argentina, were classified antigenically as follows: one serotype 1, four serotype 2, two serotype 3, and no serotype 4. Six specimens reacted with both serotype 1 and 2 MAbs, and viruses in six specimens probably belonged to other serotypes because they reacted only with a VP7 common epitope MAb. Two porcine rotavirus fecal samples found to contain both serotype 1 and 2 viruses by the MAb-based test and one found to contain a serotype 2 virus were grown in tissue culture. When plaque-purified preparations of these tissue culture-adapted viruses were analyzed in the serotyping ELISA, the C60 and C86 preparations reacted only as serotype 1 viruses, indicating that the original fecal samples, which showed multiple VP7 reactivities, were heterogeneous and apparently contained two types of viruses. Testing of plaque-purified C134 virus confirmed its serotype 2 reactivity. The MAb-based serotype designations of these viruses also were confirmed by using a neutralization immunoperoxidase focus reduction assay. This is the first report of the occurrence of serotype 1 and 2 rotaviruses in animals. The MAbs originally developed to serotype human rotaviruses can be utilized to type animal rotaviruses.

Animals↗
PubMed1990

Nucleotide sequence of VP4 and VP7 genes of human rotaviruses with subgroup I specificity and long RNA pattern: implication for new G serotype specificity.

We sequenced the genes coding for the two neutralization proteins, VP4 and VP7, of human rotavirus strains L26 and L27 with subgroup I specificity but the long RNA pattern. The deduced VP7 amino acid sequence of strains L26 and L27 showed a low homology (73.6 to 81.9%) to those of rotavirus strains of the established serotypes. This finding, together with the previous serological characterizations, suggests that the VP7 (G) serotype of the L26 and L27 strains is distinct from those of strains of the previously established serotypes. In contrast, the VP4 sequences of the L26 and L27 strains were quite similar to those of virulent serotype 2 strains (DS-1, S2, and RV-5).

Amino Acid Sequence↗
PubMed1990

Determination of subgroup, serotype and electropherotype of human rotaviruses obtained from diarrheic patients in the Philippines.

A total of 40 rotavirus specimens obtained from infants with acute gastroenteritis in Metro Manila, Philippines, were analyzed for their subgroup and serotype specificities and RNA electropherotype. A half (50%) of the specimens contained subgroup I human rotaviruses (HRVs), while 45% of them contained subgroup II HRVs. All subgroup II HRVs were determined to be serotype 1, except one specimen belonging to serotype 3, by an enzyme-linked immunosorbent assay with serotype specific monoclonal antibodies. However, serotype specificity of all subgroup I specimens failed to be determined. In polyacrylamide gel electrophoresis, RNAs extracted from HRVs of either subgroup showed long electropherotype. At least two electropherotypes were distinguished in subgroup I viruses, while five different patterns of viral RNA were found in subgroup II viruses.

Child, Preschool↗
PubMed1990

Serological and genetic characterization of bovine rotaviruses in Thailand by ELISA and RNA-RNA hybridization: detection of numerous non-serotype 6 strains.

A total of 62 fecal specimens positive for rotavirus were collected from diarrheic cows in Thailand in 1988 and 1989. The antigenic properties of rotaviruses in stool were examined by enzyme-liked immunosorbent assays using specific monoclonal antibodies directed at VP4, VP6 or VP7: all the bovine rotavirus strains were determined as subgroup I; none of the strains were reactive with serotype 6-specific monoclonal antibody; and different reactivities of the bovine strains with two anti-VP4 monoclonal antibodies were observed. Polyacrylamide gel electrophoresis of viral RNA exhibited three different RNA electropherotypes. In RNA-RNA hybridization experiments using cell culture-adapted three strains as well as a reference bovine strain (NCDV), RNA from the Thai bovine strains showed very low homology to that from NCDV; only three or four RNA segments were hybridized between the RNAs from Thai samples and NCDV. These results suggested that bovine rotaviruses isolated in Thailand are serologically and genetically distinct from a reference serotype 6 bovine strain, NCDV.

Animals↗
PubMed1989

Unusual human rotavirus strains having subgroup I specificity and "long" RNA electropherotype.

During an epidemiological study of human rotavirus in Metro Manila, Philippines, we found 20 unusual strains which belong to subgroup I but have "long" RNA pattern typical of subgroup II human rotavirus. The RNA patterns of the 20 strains were classified into four groups though they were very similar to each other. Four strains, designated L4, L26, L27, and L34 were isolated in MA104 cells from stool specimens. They possessed subgroup I specificity and long RNA pattern identical to that of the viruses in the original stool samples. The serotype specificity of these strains could not be determined by either enzyme-linked immunosorbent assay (ELISA) or neutralization test, while one strain (L27) was neutralized by serotype 2-specific anti-VP4 monoclonal antibody. These strains were suggested to have an unusual antigenicity on VP7.

Electrophoresis, Polyacrylamide Gel↗
PubMed1989

Survey of human rotavirus serotypes in different locales in Japan by enzyme-linked immunosorbent assay with monoclonal antibodies.

To investigate the relative frequency of individual human rotavirus serotypes prevailing in Japan, 562 stool specimens collected from patients with rotavirus gastroenteritis between November 1986 and March 1988 in seven districts were examined by an enzyme-linked immunosorbent assay (ELISA) with serotype 1-, 2-, 3-, and 4-specific monoclonal antibodies. Serotype 1 was the predominant serotype in the winter of 1986-1987; however, both serotypes 1 and 2 were detected frequently in the winter of 1987-1988. The results showed the relative frequency of individual serotypes by locale and the yearly change in the prevalence of each serotype in the same area. The result of subgroup specificity of rotavirus obtained by using ELISA with subgroup I- and II-specific monoclonal antibodies confirmed the general finding that rotavirus strains having subgroup I specificity are serotype 2 and those having subgroup II specificity are either serotype 1, 3, or 4. Unusual strains having both subgroup I and II specificity or neither specificity and strains presumed to represent new serotypes were also found.

Adolescent↗
PubMed1989

Dot hybridization assay for distinction of rotavirus serotypes.

We have developed a hybridization assay that permits distinction of rotavirus serotypes 1, 2, 3, and 4. The serotype of rotaviruses from stool samples or tissue culture was recognized by hybridization of specific probes to (i) blots of viral double-stranded RNAs electrophoresed in agarose gels (Northern blots) or (ii) heat-denatured double-stranded RNAs directly dotted on nylon membranes. The probes consisted of 32P-labeled cDNA synthesized by reverse transcription of in vitro derived rotavirus mRNA from rotavirus serotypes 1 to 4. To prepare these probes, mRNAs were primed with a 17-mer nucleotide common to all four serotypes whose sequence is complementary to bases 375 to 391 of the rotavirus gene encoding the VP7 glycoprotein (gene 8 or 9 depending on the rotavirus strain). The resulting downstream transcripts encompassed areas of major sequence divergence among the four serotypes. Hybridization at high stringency (50 degrees C, 50% formamide, 4 x SSC [1 x SSC is 0.15 M NaCl plus 0.015 M sodium citrate]) was performed for 16 to 48 h. Autoradiograms of the washed membranes allowed recognition of the rotavirus serotype present in the blotted or dotted specimens since each of them hybridized preferentially to one of the four probes. Twenty-four laboratory specimens and 103 clinical specimens from Washington, D.C., Venezuela, and Chile were "serotyped" with this assay. The results were similar to those obtained with a monoclonal antibody serotyping assay.

Animals↗
PubMed1989

Characterization by enzyme-linked immunosorbent assay using subgroup- and serotype-specific monoclonal antibodies of human rotavirus obtained from diarrheic patients in Bangladesh.

By enzyme-linked immunosorbent assay with group A-, subgroup-, and serotype-specific monoclonal antibodies (MAbs), we tested 414 stool specimens collected from pediatric and adult patients hospitalized with acute gastroenteritis between January and June 1988. Of 414 specimens tested, 124 (30%) were positive for group A rotavirus. The subgroup was determined in 110 specimens (88.7%); 16.1% were subgroup I, and 72.6% were subgroup II. Two specimens reacted with both subgroup I- and subgroup II-specific MAbs. Serotype determinations showed that serotype 1 (38.4%) was predominant over serotypes 2 (28.2%), 3 (2.5%), and 4 (23%). Three specimens reacted with more than one serotype-specific MAb. While the frequency of serotype 1 was highest in the two hospitals in Mymensingh, serotype 2 was most prevalent in one hospital in Dhaka. All human rotavirus strains with subgroup I and serotype 2 specificities showed a short electropherotype, and all but one strain with subgroup II and serotype 1, 3, or 4 specificities exhibited a long electropherotype.

Acute Disease↗
PubMed1989

Subgroup and serotype distributions of human, bovine, and porcine rotavirus in Thailand.

The subgroup and serotype specificities of human, bovine, and porcine group A rotaviruses in stool specimens collected in Thailand were examined by an enzyme-linked immunosorbent assay by using subgroup- and serotype-specific monoclonal antibodies. A clear yearly change was observed in the serotype distribution of human rotavirus. Between 1983 and 1984, serotype 4 was the most prevalent, while the highest frequency of serotype 2 was found between 1987 and 1988. All the bovine and porcine rotaviruses examined showed subgroup I specificities and long RNA patterns. It was of note that serotype 3 porcine rotaviruses were found at a high frequency.

Animals↗
PubMed1989

Complete nucleotide sequence of the gene encoding VP4 of a human rotavirus (strain K8) which has unique VP4 neutralization epitopes.

In our previous study (K. Taniguchi, Y. Morita, T. Urasawa, and S. Urasawa, J. Virol. 62:2421-2426, 1987) in which the cross-reactive neutralization epitopes on VP4 of human rotaviruses were analyzed, one strain, K8, was found to bear unique VP4 neutralization epitopes. This strain, which belongs to subgroup II and serotype 1, was not neutralized by any of six anti-VP4 neutralizing monoclonal antibodies which reacted with human rotavirus strains of serotypes 1, 3, and 4 or serotypes 1 through 4. We determined the complete nucleotide sequence of the gene encoding VP4 of strain K8 by primer extension. The VP4 gene is 2,359 base pairs in length, with 5' and 3' noncoding regions of 9 and 25 nucleotides, respectively. The gene contains a long open reading frame of 2,325 bases capable of coding for a protein of 775 amino acids. When compared with those of other human rotaviruses, VP4 of strain K8 had an insertion of one amino acid after residue 135, as found in simian rotavirus strains, and in addition, it had a deletion of one amino acid (residue 575). The amino acid homology of VP4 of strain K8 and those of other virulent human rotaviruses was only 60 to 70%. This was unusual, since over 90% VP4 homology has been found among the other virulent human rotavirus strains. In contrast, the VP7 amino acid sequence of the K8 strain was quite similar (over 98% homology) to those of other serotype 1 human rotaviruses. Thus, the K8 strain appears to have a unique VP4 gene previously not described.

Amino Acid Sequence↗
PubMed1988

Two sequential outbreaks of rotavirus gastroenteritis: evidence for symptomatic and asymptomatic reinfections.

In two sequential outbreaks of rotavirus gastroenteritis that occurred in a kibbutz in southern Israel (the Negev), 32 persons (9% of the population) were ill in the first and 45 (13% of the population) in the second. Excretion of virus, changes in titers of rotavirus-specific serum IgG, or both implicated rotavirus in 72% of the illnesses in outbreak 1 and in 56% of the illnesses in outbreak 2. In both outbreaks the age-specific morbidity rate decreased with increasing age. Half (six of 12) of the children six to 27 months of age who were ill with rotavirus in outbreak 1 were ill with rotavirus again in outbreak 2, whereas two were asymptomatically infected; older children who were ill in outbreak 1 were not ill in outbreak 2. Serotype determination by enzyme-linked immunosorbent assay using monoclonal antibodies to VP7 implicated a serotype 3 virus in outbreak 1 and a serotype 1 virus in outbreak 2.

Adolescent↗
PubMed1988

Analysis of serotype-specific neutralization epitopes on VP7 of human rotavirus by the use of neutralizing monoclonal antibodies and antigenic variants.

We analysed serotype-specific antigens of human rotavirus (HRV) by the use of neutralizing monoclonal antibodies (N-MAbs). The reactivity patterns of 12 serotype-specific N-MAbs against 15 HRV strains in a neutralization test revealed great intraserotypic antigenic variation especially those belonging to serotypes 2,3 and 4. On the basis of the protein specificity of the antibodies examined, it was suggested that whereas serotype 2-specific neutralization epitopes were present on both VP3 and VP7 outer capsid proteins, serotype 1-, 3- and 4-specific neutralization epitopes were located on VP7. Serotype-specific neutralization epitopes on VP7 of the serotype 1 HRV KU strain were analysed further using mutants of the KU strain resistant to different serotype 1-specific N-MAbs. The result suggested the presence of at least five operationally overlapping neutralization epitopes on VP7 of the KU strain, which collectively constituted a single large neutralization domain.

Antibodies, Monoclonal↗
PubMed1988

Validity of an enzyme-linked immunosorbent assay with serotype-specific monoclonal antibodies for serotyping human rotavirus in stool specimens.

We recently developed a method for serotyping human rotavirus (HRV) by an enzyme-linked immunosorbent assay with HRV serotype-specific neutralizing monoclonal antibodies (ELISA serotyping). In the present study this method was compared with the fluorescent focus neutralization test with serotype-specific rabbit antisera (NT serotyping) in the sensitivity and specificity of the test. Direct serotyping of HRVs which were contained in stool specimens indicated that while only 37% of the samples were successfully serotyped in NT, 78% of the samples could be serotyped in ELISA. Regarding the samples whose serotype could be determined in the two tests, the assigned serotypes were identical in both tests. The results obtained indicated the utility of ELISA serotyping in clinical and epidemiologic studies of HRV infection.

Antibodies, Monoclonal↗
PubMed1988

Relative frequencies of rotavirus serotypes 1, 2, 3, and 4 in Venezuelan infants with gastroenteritis.

We have used a recently developed monoclonal antibody enzyme-linked immunosorbent assay (K. Taniguchi, T. Urasawa, Y. Morita, H. B. Greenberg, and S. Urasawa, J. Infect. Dis. 155:1159-1166, 1987) for serotyping rotaviruses recovered from 134 Venezuelan infants over a period of 15 months. One hundred and nine of the specimens were typed with the following distribution: serotype 1, 48%; serotype 2, 16%; serotype 3, 22%; and serotype 4, 14%. Three specimens reacted with two different monoclonal antibodies. In addition, 6 specimens (5%) containing enough outer capsid antigen could not be typed; partial RNA sequence analysis of the glycoprotein gene from three of these six strains failed to reveal sequence differences with prototype strains that could be serotyped with the monoclonal antibodies. Variations in the recovery rates of the different serotypes were observed. Serotypes 2, 3, and 4 predominated at the beginning of the study, and serotype 1 predominated at the end of the study. Diarrheal illness appeared to be more prolonged in infants shedding rotavirus serotypes 1 and 3 than in those shedding serotypes 2 and 4.

Base Sequence↗