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R F Bishop

Publications and source records attributed to R F Bishop.

At least 19 recordsLinked to original sources
PubMed1992

Typing of human group A rotavirus with alkaline phosphatase-labeled oligonucleotide probes.

Rotavirus (RV) in stools of children less than 1 year of age with diarrhea in Bangkok in 1989 were serotyped by monoclonal enzyme immunoassay (MEIA). RNA extracted from these specimens was tested for hybridization with alkaline phosphatase (AP) and 32P-labeled oligonucleotides constructed from the nucleotide sequences of VP7 of human G types 1 (HuG1Ac), 2 (HuG2Ac), 3 (HuG3Ac), and 4 (HuG4Ac). Of 148 specimens that contained RV, 72% (106/148) hybridized with RV G type specific AP-labeled oligonucleotides compared to 47% (70/148) that were serotyped by MEIA (P less than 0.001). Of 68 specimens that contained only one VP7 serotype (G-type), as identified by MEIA, 94% (16/17) of G1, 90% (27/30) of G2, 57% (4/7) of G3, and 36% (5/14) of G4 RV hybridized with the AP-labeled HuG1Ac, HuG2Ac, HuG3Ac, and HuG4Ac oligonucleotides, respectively. The probes for G1, 2, 3, and 4 RV were specific for each G type. The results of hybridizing specimens with 32P- and AP-labeled oligonucleotides were similar. After transcription and amplification of cDNA of gene 9, AP-labeled RV G type specific oligonucleotides hybridized with 90% (134/148) of RV specimens. The high sensitivity of these nonimmunological techniques could be of value in identifying G types of RV during vaccine trials.

Alkaline Phosphatase↗
PubMed1992

Severity of rotavirus infection in relation to serotype, monotype and electropherotype.

The aim of this study was to determine whether the severity of symptoms associated with rotavirus infection was related to the serotype of the infecting virus. Severity of clinical symptoms in 108 children admitted to hospital for treatment of rotavirus diarrhoea was retrospectively assessed using a scoring system for frequency and duration of vomiting and diarrhoea, degree of fever, acidosis and dehydration, and presence of electrolyte imbalance. Children were 6-30 months old and were fully weaned at onset of symptoms prior to admission to hospital. No other enteric pathogens were detected during the course of the illness. Serotypes and monotypes were identified using a panel of monoclonal antibodies. Gel electrophoresis of rotavirus RNA was performed to determine electropherotypes. Children surveyed were infected with serotype 1 (47), serotype 2 (15) or serotype 4 (46) rotaviruses. Comparisons of severity of clinical symptoms according to infecting serotype revealed no statistically significant differences between serotype 1, 2 or 4 infections. In addition, no differences were detected between different rotavirus strains within each serotype (as judged by electropherotype) including monotypes 1a or 1c. This study failed to reveal differences in virulence between rotavirus strains of different VP7 serotypes infecting young children.

Child, Preschool↗
PubMed1992

Comparisons of rotavirus VP7-typing monoclonal antibodies by competition binding assay.

Three sets of neutralizing monoclonal antibodies (MAbs) used to type the outer capsid protein VP7 of four group A rotavirus serotypes (1 through 4) were compared in competition immunoassays. Reciprocal competition was observed for each of the VP7 type 2-, 3-, and 4-specific MAbs. The VP7 type 1 MAbs exhibited variable competition patterns with other VP7 type 1 MAbs. MAb RV4:3, which has been used to recognize antigenic variants within VP7 type 1 strains, showed reciprocal competition with the four VP7 type 3 MAbs (RV3:1, YO-1E2, 4F8, and 159) using a VP7 type 3 virus (SA11) as antigen. MAb 2C9, also prepared against VP7 type 1, reacted with VP7 type 3 strains and competed with a VP7 type 3 MAb, 159, using RRV as antigen. Use of the different sets of VP7 type-specific MAbs in the enzyme-linked immunosorbent assay permitted the recognition of six antigenic variants within VP7 types 1, 2, and 3 among specimens whose VP7 type could not be determined previously with only one set of typing MAbs. These results demonstrate differences of typing ability among these VP7-specific MAbs and emphasize the need to improve the sensitivity of typing systems by incorporating panels of MAbs reacting with several neutralizing epitopes.

Antibodies, Monoclonal↗
PubMed1992

Role of coproantibody in clinical protection of children during reinfection with rotavirus.

Rotavirus is the major cause of severe, dehydrating infantile gastroenteritis. Infection is limited to the gut, but the relative roles of serum and secretory copro-immunoglobulin A (IgA) in protection are unclear. Specific copro-IgA is predictive of duodenal antirotaviral IgA and correlates with virus-neutralizing coproantibody. Copro-IgA conversion is a more sensitive marker of rotavirus reinfection than seroconversion. We measured rotavirus reinfections by copro-IgA conversion prospectively in 35 children recruited at a time of severe rotavirus illness. The children were followed up longitudinally for 14 to 31 months to determine whether high coproantibody levels correlated with clinical protection against rotavirus disease. Ninety-four percent of the children experienced reinfection, and 38% developed persistent elevations in specific copro-IgA termed plateaus. Plateau children had a higher mean annual rate of rotavirus infection and a lower ratio of symptomatic to total number of rotavirus reinfections than did nonplateau children. The annual rates of rotavirus infection and disease were significantly higher outside the plateau than inside it in children experiencing antirotavirus copro-IgA plateaus. Frequent rotavirus infection of children appears to stimulate production of a specific copro-IgA plateau which correlates with protection against an excess of infection and symptomatic disease.

Age Factors↗
PubMed1991

Reactivities of serotyping monoclonal antibodies with culture-adapted human rotaviruses.

Rotaviruses collected in Bangladesh during 1985 to 1986 were culture adapted and used in a comparative serotyping study with three groups of monoclonal antibodies, all of which reacted with the major neutralization protein (VP7) of serotype 1, 2, 3, or 4. The goals were to determine which monoclonal antibodies most accurately predicted the serotype and why large variations in serotyping efficiencies have occurred with these monoclonal antibodies in previous studies. The 143 rotavirus isolates used in this study belonged to 69 different electropherotypes; and 44, 23, 21, and 55 isolates were identified as serotype 1 through 4, respectively, by neutralization with serotype-specific hyperimmune antisera. Serotyping specificity by enzyme-linked immunosorbent assay with monoclonal antibodies was 100% consistent with results found by neutralization with polyclonal antisera, but large differences were observed in the sensitivities of the different monoclonal antibodies. Monoclonal antibodies 5E8 (serotype 1), 1C10 (serotype 2), 159 (serotype 3), RV3:1 (serotype 3), ST-3:1 (serotype 4), and ST-2G7 (serotype 4) reacted with all the isolates of the corresponding serotype for which there were sufficient infectious particles. Monoclonal antibody 2F1 (serotype 2) was much less sensitive and reacted with only five serotype 2 isolates, but these were among those with the highest titers. Monoclonal antibodies RV4:2 (serotype 1), KU6BG (serotype 1), RV5:3 (serotype 2), and S2-2G10 (serotype 2), on the other hand, failed to react with between one and three isolates of the corresponding serotypes which had high titers, apparently because of epitope changes in these isolates. Effects of epitope variation were, however, most apparent with monoclonal antibodies 2C9 (serotype 1) and YO-1E2 (serotype 3), which reacted with one and no isolates of the corresponding serotypes, respectively. Cross-neutralization of escape mutants indicated that the serotype 1 monoclonal antibodies 5E8, 2C9, and RV4:2 reacted with different but probably overlapping epitopes, as did serotype 2 monoclonal antibodies 2F1, 1C10, and RV5:3, finding that were consistent with the enzyme-linked immunosorbent assay data. Because of epitope variations between rotavirus strains, serotyping with several monoclonal antibodies directed at different epitopes may increase the sensitivity of the method.

Antibodies, Monoclonal↗
PubMed1991

Epidemiology of rotavirus serotypes in Melbourne, Australia, from 1973 to 1989.

Fecal rotavirus strains collected between 1973 and 1989 from 943 children admitted with acute diarrhea to one hospital in Melbourne, Australia, were serotyped by using an enzyme-linked immunosorbent assay. The assay incorporated neutralizing monoclonal antibodies specific for VP7 of the four major human serotypes (1 through 4). A serotype could be assigned to 690 of 943 specimens (73.2%). Typeable strains comprised serotype 1 (72.5%), serotype 2 (6.8%), serotype 3 (2.9%), or serotype 4 (15.4%). Monotypes 1a and 1c comprised 52 and 44%, respectively, of serotype 1 strains. All serotypes and monotypes exhibited polymorphic genomic RNAs. Specimens reacting as mixed serotypes were rare (3.2%) and included intertypic strains (0.7%) and mixed infections (1.0%). Nontypeable strains for which an electropherotype could be determined appeared to be identical with typeable strains present concurrently in the community. Serotypes exhibited various epidemiological patterns. Serotype 1 strains were dominant except during three successive winters when 60 to 90% of the disease was caused by serotype 2. Serotype 4 strains showed an episodic pattern of appearance, recurring at peak incidence approximately every 3 years. Fecal rotavirus strains collected from 145 newborn babies housed in Melbourne obstetric hospitals between 1974 and 1986 were also serotyped. All 135 typeable strains (93.1%) belonged to serotype 3. It is hypothesized that endemic infection with serotype 3 rotaviruses in nurseries for the newborn influenced the epidemiology of rotavirus serotypes responsible for severe clinical disease in young children in the same community.

Age Factors↗
PubMed1990

Serotyping of human group A rotavirus with oligonucleotide probes.

Rotaviruses (RV) in stools of children with diarrhea in Thailand were serotyped by monoclonal enzyme immunoassay (MEIA), and RNA extracted from these specimens were tested for hybridization with oligonucleotides constructed from the nucleotide sequences of VP7 of human serotypes 1, 2, 3, and 4. Of 178 specimens that contained RV as identified with a monoclonal antibody to group A RV, 84% (149/178) hybridized with serotype-specific oligonucleotides, and 42% (74/178) were serotyped by MEIA (P less than .001). Of the 74 specimens that were serotyped by MEIA, 92% (35/38) of type 1, 97% (34/35) of type 2, and the one type 4 RV hybridized with the HuG1Ac, HuG2Ac, and HuG4Ac oligonucleotides, respectively. RV strains identified in children in Thailand in 1987 and 1988 to which a serotype could be assigned by either method were either type 1, type 2, or, less often, type 4. Testing RV for hybridization with oligonucleotides for genes encoding VP7 is an alternate method of determining RV serotypes.

Antibodies, Monoclonal↗
PubMed1990

Comparison of rotavirus immunoglobulin A coproconversion with other indices of rotavirus infection in a longitudinal study in childhood.

In order to determine the sensitivity and reliability of antirotaviral fecal immunoglobulin A (IgA) as an indicator of rotavirus reinfection, the antibody responses to rotavirus of 44 infants with severe rotavirus gastroenteritis recruited on admission to a hospital were studied. Feces were collected daily during hospitalization and weekly thereafter, and sera were obtained every 4 months, for 6 to 32 months (median, 17 months). Antirotaviral IgG, IgA, and IgM were measured by enzyme immunoassay in all samples. Rotavirus antigen, rotavirus-neutralizing antibody, and total IgA were measured in feces. The results showed that use of an IgA index (ratio of specific IgA to total IgA) was unnecessary to identify copro-IgA conversion to rotavirus. The other markers of rotavirus infection tested showed a high level of predictive accuracy of coproconversion in rotavirus-neutralizing antibody. Copro-IgM, serum IgM, and virus in feces were insensitive measures of neutralizing antibody coproconversion. Seroconversion in IgG or IgA was detected in 46% of neutralizing coproconversions. The most sensitive marker, present in 92% of neutralizing coproconversions, was antirotaviral fecal IgA conversion. This correlation of fecal IgA with fecal neutralizing antibody suggests that coproconversions in IgA represent true elevations in antirotaviral IgA with neutralizing capacity. A coproconversion in IgA appears to indicate genuine rotavirus infection. Copro-IgA conversions in feces collected weekly are likely to be more sensitive markers of rotavirus reinfection than are seroconversion and virus detection combined in epidemiological studies of acute diarrhea in children and in rotavirus vaccine trials.

Antibodies, Viral↗
PubMed1990

Homotypic serum antibody responses to rotavirus proteins following primary infection of young children with serotype 1 rotavirus.

The class-specific antibody responses to serotype 1 rotavirus structural proteins were examined by immunoblotting with sera obtained from young children hospitalized with acute rotavirus diarrhea caused by serotype 1. All were believed to be primary infections. Three consecutive samples were obtained from 16 patients during the acute and convalescent phases of the disease and then approximately 4 months later. Immunoglobulin G (IgG)-class antibody responses to two inner capsid proteins (VP2 and VP6) and to the major homologous outer capsid protein (VP7) were detected in all patients. Antibody responses to VP6 were rapid, increased in intensity during 20 to 40 days after the onset of symptoms, and persisted for more than 4 months. Responses to VP2 and VP7 were more delayed, were maximal in convalescent-phase sera, and decreased markedly in intensity 4 months after the onset of symptoms in the majority of children. Two patients with evidence of mixed infection showed persisting high levels of antibody to VP7. Responses to the outer capsid protein VP4 were detected in 67% of patients, peaked at 20 to 40 days after the onset of symptoms, and were no longer detected at 4 months in the majority of patients. It is likely that the immunoblotting technique underestimated responses to VP4. Acute- and convalescent-phase sera (known to contain antirotavirus IgM or IgA measured by enzyme immunoassay) were also examined by immunoblotting. IgM- and IgA-class antibody responses to viral proteins VP2, VP4, and VP7 appeared to be qualitatively identical to those observed for IgG in the same serum samples.

Antibodies, Viral↗
PubMed1979

Diarrhea and rotavirus infection associated with differing regimens for postnatal care of newborn babies.

Surveillance of 2,041 babies born during 4 winter months in one obstetric hospital in Melbourne, Australia, showed that 215 developed acute diarrhea during the first 2 weeks of life. Babies requiring special care from birth had a high incidence of sporadic diarrhea (36%). The incidence of diarrhea among healthy full-term babies was low if they were "rooming-in" with their mothers (2 to 3%) but high if they were housed in communal nurseries (29%). The most important factor influencing incidence of diarrhea was proximity to other newborn babies and frequency of handling by related adults. Breast feeding did not always protect babies from diarrhea. Excretion of rotaviruses was temporally retlated to diarrhea in 61 to 76% of healthy full-term babies and in 44% of babies requiring special care. Other eneteric pathogens, including enerotoxigenic Escherichia coli, were occasionally isolated. Calculation of the ratios of symptomatic to asymptomatic infection suggests that babies requiring special care are much more likely to develop symptomatic illness after rotavious infection than are full-term babies.

Australia↗
PubMed1978

Enterotoxin-producing Escherichia coli and diarrhoea among newborn babies in Darwin.

Specimens of Escherichia coli O119B14 isolated from 13 newborn babies during an outbreak of diarrhoea in Darwin Hospital were tested for their ability to produce enterotoxin. Of the 13 isolates tested none was found to produce heat-labile enterotoxin. However, isolates from two of the five diarrhoeal faecal specimens were found to produce moderate levels of heat-stable enterotoxin (ST). It seems like that ST-producing strains of E. coli were responsible for the diarrhoea.

Australia↗
PubMed1978

Pattern of shedding of two noncultivable viruses in stools of newborn babies.

Noncultivable viruses have been associated with diarrhea affecting newborn babies in obstetric hospital nurseries. Persisting infection in a special care nursery in Melbourne, Australia, permitted a study of the pattern of excretion of these viruses. Ten babies admitted to the nursery within 2 hr of birth were randomly selected for prospective study. Feces were collected daily for 14 days and were examined by electron microscopy. All ten babies excreted detectable amounts of duovirus (rotavirus, HRVL agent, IGV) for at least 1 day. Age at onset of excretion varied from 2 to 13 days. Eight of the ten babies developed diarrhea. Excretion of duovirus preceded the onset of diarrhea by 12--72 hr and persisted for at least 3 days. Seven of the ten babies also excreted detectable amounts of 28-nm virus-like particle for 3--8 days. The identity of this particle is unknown. Morphologically it resembles Norwalk agent and "astrovirus." Excretion of this 28-nm particle coincided with symptoms of diarrhea in four babies, all of whom were also excreting duovirus. It is concluded that most newborn babies admitted to a nursery where duovirus infection is endemic will excrete this virus at least once during the first 2 weeks of life. Excretion of virus particles will either precede development of diarrhea or be asymptomatic. Selective isolation of babies with diarrhea is thus unlikely to control spread of duovirus infection within a hospital nursery.

Australia↗
PubMed1978

Noncultivable viruses and neonatal diarrhea: fifteen-month survey in a newborn special care nursery.

During a 15-month period of surveillance, diarrhea developed in 257 of 913 babies (28%) admitted within 2 hours of birth to a special care nursery in Melbourne, Australia. Diarrhea was seasonal, affecting a maximum of 43% of babies admitted during one winter month (July) and a minimum of 13% of babies admitted during one summer month (December). Diarrhea was no more frequent nor more severe in babies of very low birth weight or of very early gestational age. Two noncultivable viruses were located by electron microscopy in feces from babies with or without diarrhea. Excretion of a reovirus-like particle (rotavirus, duovirus, human reovirus-like agent, infantile gastroenteritis virus) was temporally related to diarrheal symptoms. Asymptomatic infection with this virus also occurred. A 28-nm virus-like particle was excreted by some babies, but it could not be implicated on epidemiological grounds in the etiology of the diarrhea. Rotavirus infection may be an important cause of endemic diarrhea in nurseries for the newborn. Infection may be difficult to control or eradicate, since it is often asymptomatic and may be influenced by infection in the community at large.

Australia↗
PubMed1977

Complement-fixing antibody response to rotavirus infection.

A human rotavirus complement-fixing (CF) antigen, prepared by purification of large volumes of fluid feces collected from children with winter diarrhea, was used to study the development and persistence of antibody in children with diarrhea and the prevalence of rotavirus antibody in Melbourne. In children with diarrhea, antibody rises were detectable within 4 to 6 weeks of the onset of illness, and the titers usually remained elevated for the next 1 to 2 years. CF antibody did not develop in two children with proven rotavirus infection aged less than 6 months, an age at which poor CF responses to other viruses have also been observed. A study of CF antibody levels in the general community showed that in Melbourne, most children have been infected with human rotavirus by the age of 3 years.

Age Factors↗
PubMed1976

Is lactase the receptor and uncoating enzyme for infantile enteritis (rota) viruses?

Rotaviruses are now regarded as important causes of diarrhoea in man, cattle, pigs, mice, and possibly other animals. Characteristically, disease occurs in newborn and young animals, and infection seems limited to the differentiated gut epithelial cells. The major surface polypeptide of the calf scours rotavirus is glycosylated, and highly purified beta-galactosidase (lactase) interacts with the virus in vitro causing removal of the outer shell of the capsid (uncoating). It is suggested that lactase present in the brush border of the intestinal epithelial cell performs a similar function in vivo by acting as a combined receptor and uncoating enzyme for the rotavirus. This hypothesis is consistent with the observations that rotaviruses seem to infect only gut epithelial cells, and that infant animals, whose lactase concentrations are generally higher than those of adult animals, seem more susceptible to rotavirus infections. Implications of the hypothesis include possible new approaches to laboratory cultivation of rotaviruses, which should be more successful in cells selected for surface lactase activity, and the suggestion that the epidemiology of human rotavirus infections may be influenced by the fact that different ethnic groups have different lactase levels (and hence lactose intolerance) in adulthood.

Acute Disease↗
PubMed1976

New virus associated with diarrhoea in neonates.

Since December, 1974, there has been an increase in the incidence of acute diarrhoea in the neonatal nurseries of five Melbourne metropolitan hospitals. Four of these have had epidemics, and the incidence of endemic diarrhoeal disease has increased. Extracts of faeces from 148 patients from the five hospitals were examined by electron microscopy. "Duovirus" particles were detected in 82 of these extracts, including at least one from each hospital. No bacterial pathogens were isolated. It seems likely that "duovirus" is an important cause of sporadic and epidemic acute diarrhoeal disease in neonates. It is important to note that the absence of a recognized bacterial pathogen does not exclude an infective cause, especially when sugar intolerance is present. Appropriate measures to minimize the spread of infection must be employed.

Acute Disease↗