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  • 11
    Electronic Resource
    Electronic Resource
    Springer
    European journal of pediatrics 155 (1996), S. 954-958 
    ISSN: 1432-1076
    Keywords: Nosocomial infection ; Rotavirus gastro-enteritis ; SDS-PAGE ; Serotyping ELISA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Faecal samples were collected from patients with gastro-enteritis during two winter seasons on a paediatric ward. Three outbreaks of nosocomial rotavirus gastro-enteritis were identified by latex agglutination and the virus strains were characterized by polyacrylamide gel electrophoresis of the genome nucleic acid and by subgrouping and serotyping enzyme-linked immunosorbent assays (ELISA). One outbreak was caused by serotype 1 rotavirus, one by serotype 2 and the remaining outbreak was caused by a mixture of serotypes 1 and 4. Identical electrophoretic patterns of the rotavirus genome in each outbreak combined with the ELISA results indicate that these three outbreaks were hospital-acquired cases. The index cases in the three outbreaks were community-acquired and one of two index cases in the second outbreak was hospital-acquired. On each occasion, susceptible roommates were easily infected from the index cases and then cross-infection occurred in the paediatric ward. Possible vehicles were the medical staff, especially doctors, parents of infected patients and infected patients who were moved to other rooms. One patient who had been treated with a series of antitumour therapies excreted rotaviruses in faeces for a long time period and probably played a role as a source.of the outbreak. Moreover, some patients still excreted rotaviruses in their normal stool 1 week after recovery from gastro-enteritis. These findings indicate that continual examination of stool samples for rotaviruses until they are negative may be important to prevent the spread of rotavirus infection.
    Type of Medium: Electronic Resource
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  • 12
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary We studied the competitive growth among SA11-L2(G3) and its single-human VP7 gene-substitution reassortants SA11-L2/KU-R1(G1) and SA11-L2/DS1-R1(G2), which have the genetic background of SA11-L2, during sequential passages after mixed infection. When the same infectious units (m.o.i. of 5 p.f.u./cell) of SA11-L2 and a reassortant SA11-L2/KU-R1 were inoculated onto and passaged in MA104 cells, 88% of the virus clones isolated from the culture fluid at the 3rd passage belonged to G3, and all the clones from the 10th passage had G3 specificity. Even when SA11-L2/KU-R1 with titer 10 times higher than that of SA11-L2 was used in the coinfection, the predominance of clones with G3-VP7 was observed. Although G2 clones slightly surpassed G1 clones in number in the mixed culture of SA11-L2/KU-R1 and SA11-L2/DS1-R1, G3 clones predominated in the virus progeny from a mixed culture infected with the same titers of SA11-L2, SA11-L2/KU-R1, and SA11-L2/DS1-R1. However, no significant difference in viral growth was detected among SA11-L2 and the two reassortants.
    Type of Medium: Electronic Resource
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  • 13
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Four group A human rotaviruses having antigenic specificity of subgroup I and ‘long’ RNA electropherotype were isolated in MA104 cell cultures. Cross-neutralization tests with hyperimmune antisera suggested that they are serologically distinct from the six previously recognized human rotavirus serotypes.
    Type of Medium: Electronic Resource
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  • 14
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary VP7-specific neutralizing monoclonal antibodies (N-MAbs) to serotype 3 human rotavirus were produced to analyze serotype 3-specific and cross-reactive neutralization epitopes on VP7. On the basis of the reactivity patterns in neutralization tests with various human and animal strains, a total of 10 N-MAbs could be classified into four groups; five antibodies specific to serotype 3 were divided into two groups, and five antibodies consisted of two groups which are cross-reactive with strain 69M (serotype 8) or strain WI61 (serotype 9). Seven N-MAbs showed the same reactivity patterns to the virus strains in both neutralization tests and enzyme-linked immunosorbent assay (ELISA), while three N-MAbs specific to serotype 3 in neutralization showed a cross-reactivity with the serotype 8 strain in ELISA. Neutralization-resistant mutants of serotype 3 strains P and YO were selected by the N-MAbs. Cross-neutralization tests between the mutants and the MAbs indicated the presence of two serotype-specific (S1 and S2) and three cross-reactive (C1, C2, and C3) epitope groups. S1, S2, and C3 epitope groups overlapped operationally each other, and the S1 epitope group had an overlapping with the C1 epitope group. However, C2 epitope group identified by the MAbs which neutralized serotypes 3 and 9, had no operational overlapping with any other epitope groups.
    Type of Medium: Electronic Resource
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  • 15
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary A non-neutralizing monoclonal antibody (YO-60) against human rotavirus was found to be directed to VP2 (90,000-dalton protein), one of the two major components of the inner capsid. The reactivity patterns of the YO-60 antibody were very similar, though not identical, to those of subgroup II-specific YO-5 monoclonal antibody directed to VP6 (42,000-dalton protein), the other major component of the inner capsid. These results indicated the possible presence of a subgroup-specific antigen on VP2 in addition to the one on VP6.
    Type of Medium: Electronic Resource
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  • 16
    Electronic Resource
    Electronic Resource
    Springer
    Archives of virology 139 (1994), S. 209-215 
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Antigenic and genomic properties of equine rotavirus strain CH3 isolated in Japan were studied by cross-neutralization tests and nucleotide sequence determination of the VP4 and VP7 genes. It was shown that the strain CH3 belongs to G14 and shares VP4 genotype with strain H2.
    Type of Medium: Electronic Resource
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  • 17
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary We previously found the preferential selection of VP7 gene from a parent rotavirus strain SA11 with G serotype 3 (G3) in the sequential passages after mixed infection of simian rotavirus SA11 and SA11-human rotavirus single-VP7 gene-substitution reassortants with G1, G2, or G4 specificity. However, it has not been known whether or not VP7 genes derived from other strains with G3 specificity (G3-VP7 gene) are preferentially selected in the genetic background of SA11. To address this question, mixed infections followed by multiple passages were performed with a reassortant SA11-L2/KU-R1 (SKR1) (which possesses VP7 gene derived from G1 human rotavirus KU and other 10 genes of SA11 origin) and one of the five G3-rotaviruses, RRV, K9, YO, AK35, and S3. After the 10th passage, selection rates of SA11-L2/KU-R1 gene 9 (G1-VP7 gene) and gene 5 (NSP1 gene) reduced considerably (0 to 20.4%) in the clones obtained from all the coinfection experiments, while all or some of other segments were preferentially selected from SKR1 depending on the pairs of coinfection. When viral growth kinetics was examined, SKR1 exhibited better growth and reached a higher titer than any G3 viruses. Although the generated reassortants with VP7 gene and NSP1 gene derived from G3 viruses showed almost similar growth kinetics to that of SKR1 during the first 20 h of replication, the titers of these reassortants were higher than that of SKR1 after 36h postinfection. The results obtained in this study suggested that G3-VP7 gene is functionally more adapted to the genetic background of SA11.
    Type of Medium: Electronic Resource
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  • 18
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Two neutralizing monoclonal antibodies (4 C 4 and 4 F 2) against type 1 poliovirus, Mahoney strain, recognized distinct antigenic sites of the virus particles; 4 C 4 antibody bound to vertices of native and heated (56° C, 30 minutes) virus of Mahoney strain, while 4 F 2 antibody reacted with specific surface protrusions of native virus of Mahoney and Sabin strains. The difference in the location of neutralization epitopes with which the two antibodies react was confirmed in the neutralization reaction by the use of mutants resistant to 4 C 4 and 4 F 2 antibody. In immune electron microscopy, double immunodiffusion and sucrose density gradient analysis of virus-antibody complexes, the two antibodies showed a marked difference in their virus-precipitating activities. The 4 C 4 antibody recognizing vertices of the virus particle had little virus-precipitating activity. In contrast, the 4 F 2 antibody that bound to specific surface protrusions of native virus aggregated virus particle efficiently. In neutralization assays, however, the 4 C 4 antibody exhibited a slightly stronger neutralizing activity than the 4 F 2 antibody. Thus, it was suggested that the strength in precipitating activities of the two antibodies did not correlate with that in their neutralizing activities.
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  • 19
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary For selecting the neutralizing monoclonal antibodies (N-MAbs) directed to VP4 of rotavirus strain K8, which has unique VP4 neutralization epitopes, we prepared several reassortant viruses by mixed infection of two different strains K8 (serotype 1) and P (serotype 3) in vitro: three reassortant clones having VP4 of K8 and VP7 of P and four clones having VP4 of P and VP7 of K8. By using these reassortants in screening hybridomas, a N-MAb (K8-2C12) directed to strain K8-specific VP4 was obtained. The MAb K8-2C12 neutralized only K8 when tested against numerous strains of different serotypes, while in enzyme-linked immunosorbent assay this MAb reacted also with simian rotavirus SA11 (serotype 3), bovine rotavirus NCDV (serotype 6), and human rotavirus (HRV) strain 69M (serotype 8). Neutralization-resistant mutants of K8 were selected by the K8-2C12 antibody and VP4 amino acid sequences of the mutants were determined. Single amino acid substitution was detected in the three mutant clones at position 394, which is included in the major cross-reactive neutralization region identified in other rotaviruses.
    Type of Medium: Electronic Resource
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  • 20
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary In order to study the effect of selection pressure of anti-VP4 and anti-VP7 neutralizing monoclonal antibodies(N-MAbs) on reassortant formation, 424 reassortant clones were produced from mixed cultures of human rotavirus strains Wa and HN126 and their genotypes were analysed. Reassortant selection was done with four types of N-MAb: anti-VP4 MAb to Wa and anti-VP7 MAb to HN126(selection A); anti-VP4 MAb to HN126 and anti-VP7 MAb to Wa(selection B); anti-VP7 MAb to Wa(selection C); and anti-VP4 MAb to Wa(selection D). In each selection experiment, more than 100 clones were isolated, and the parental origin of RNA segments was identified by polyacrylamide gel electrophoresis. All clones isolated by selections A and B were found to be antigenic mosaic reassortants with the VP4 gene of HN126 and the VP7 gene of Wa and antigenic mosaic reassortants with the VP4 gene of Wa and the VP7 gene of HN126, respectively. Although in reassortants of both selections, RNA segments 2, 3, 5 and 6 were selected from strain Wa at considerably high rates, selection rates of RNA segments 1, 7, 8, and 11 were significantly different between selection A and B. In reassortants from selection C and D, selection rates of RNA segments 1, 3, 6, 7, 8, and 11 from Wa were significantly lower than those in selection A and B, whereas RNA segments 2 and 5 were almost exclusively selected from Wa as observed in selection A and B. These results indicated the presence of two types of nonrandom gene selection in reassortant formation, one strongly dependent on, and another irrespective of, the selection pressure with N-MAbs.
    Type of Medium: Electronic Resource
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