Epidemiology of rotavirus electropherotypes determined by a simplified diagnostic technique with RNA analysis

Dolan, Kathleen T.; Twist, E. Michael; Horton-Slight, Paula; Forrer, Christine B.; Bell, Louis M.; Plotkin, S. A.; Clark, H Fred

doi:10.1128/jcm.21.5.753-758.1985

Cited by 73 publications

(22 citation statements)

References 32 publications

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“…Finally, rotaviruses from both patients displayed atypical RNA profiles as previously described for other immunodeficient patients chronically infected with group A rotaviruses [Pedley et al, 1984;Dolan et al, 1985;Eiden et al, 19851. The extra segments of ds RNA seen in these profiles have been shown to be concatemers of lower MW genome segments [Pedley et al, 19841, but how the complicated patterns of bands arise has not previously been clear.…”

Section: Discussionmentioning

confidence: 93%

“…can occur in other forms of immune deficiency, such as agammaglobulinemic patients and infants with SCID [Saulsbury et al, 1980;Pedley et al, 1984;Dolan et al, 1985;Eiden et al, 19851. The spectrum of immunodeficient children at risk from chronic enteric virus infection is, therefore, extended.…”

Section: Discussionmentioning

confidence: 99%

“…The genomes of group A rotaviruses obtained from chronically infected immunodeficient children show atypical profiles when analyzed by polyacrylamide gel electrophoresis (PAGE) [Pedley et al, 1984;Dolan et al, 1985;Eiden et al, 19851. Some normal RNA segments are missing from the profile and various additional bands of double-stranded (ds) RNA migrate between segments 1-9.…”

Section: Introductionmentioning

confidence: 99%

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Chronic enteric virus infection in two T‐cell immunodeficient children

Wood¹,

David

Chrystie

et al. 1988

Journal of Medical Virology

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Enteric virus infections were studied in two children with congenital T-cell immunodeficiency. One patient (LC) with cartilage hair hypoplasia developed persistent diarrhea and malabsorption following acute gastroenteritis. Electron microscope (EM) examination of feces revealed excretion of rotavirus for more than 450 days with concurrent astrovirus infection for at least 225 days, associated with the persistent diarrhea. Prolonged infection with poliovirus type 2 following vaccination had previously been noted in this patient. The second patient (DT), with the CHARGE association and DiGeorge syndrome, had two episodes of loose stools. EM of fecal extracts demonstrated rotavirus excretion for at least 66 days following the initial episode. Virus-specific immune responses were assayed in these two patients. LC showed a poor serum neutralizing antibody response to polio vaccination, no detectable antibody response (by immune EM and ELISA) to rotavirus, and no detectable antibody response to astrovirus (by immune EM). Rotavirus specific cell mediated immunity was also not detectable. DT showed no detectable serum antibody response to rotavirus (by ELISA). Rotavirus isolates from both patients were found to be group A viruses and were further analyzed by polyacrylamide gel electrophoresis. Atypical genome profiles, with multiple additional bands between segments 3-7 of the normal rotavirus profile, were obtained throughout the course of each illness, including the earliest specimens available (day 41, LC; day 7, DT). These results indicate that chronic virus infection of the gut can occur in patients with T-cell immunodeficiency. Such chronic infection may be associated with persistent diarrhea and can cause considerable problems of management.

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Section: Discussionmentioning

confidence: 93%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Chronic enteric virus infection in two T‐cell immunodeficient children

Wood¹,

David

Chrystie

et al. 1988

Journal of Medical Virology

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“…The genome pattern was characterized in 10% of polyacrylamide resolving gels (with an overlayered 4% of stacking gel) using Laemmli's discontinuous buffer system without SDS [Laemmli, 1970]. The RNA specimens were run at 16 mA for 20 hr at 48C and then stained with silver as described by Dolan et al [1985].…”

Section: Genome Pattern Analysis By Pagementioning

confidence: 99%

Dominating prevalence of P[8],G1 and P[8],G9 rotavirus strains among children admitted to hospital between 2000 and 2003 in Budapest, Hungary

et al. 2005

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Group A rotaviruses are the main cause of acute dehydrating diarrhea in children, responsible for high mortality in developing countries and a significant socio-economic burden associated with treating the disease in developed countries. Two rotavirus vaccine candidates predicated on either homotypic or heterotypic protection have undergone clinical trials recently and await licensure for routine use. In anticipation of a future vaccination campaign in Hungary, the diversity of rotaviruses collected from Budapest between 2000 and 2003 were analyzed by polyacrylamide gel electrophoresis (PAGE) of the viral genome and by serotyping and genotyping of the outer capsid genes, VP7 and VP4. Among 2,763 rotavirus positive specimens available for analysis, we were able to determine the electropherotype of 2,227, and, of these, 1,517 (68.1%) were subjected to G typing and 1,173 (52.7%) were subjected to P typing. We successfully G typed 1,481 (97.6%) and P typed 1,130 (96.3%) strains, respectively. A total of six G types (G1, 50.2%; G2, 2.2%; G3, 1.7%; G4, 5.8%; G6, 0.6%; and G9, 34.4%) and four P types (P[4], 3.0%; P[6], 0.7%; P[8], 89.9%; and P[9], 1.7%) were identified in nine individual combinations (P[8],G1; P[4],G2; P[8],G3; P[8],G4; P[8],G9; P[6],G4; P[4],G1; P[9],G3; and P[9],G6). The prevalence of VP7 and VP4 specificities varied from year to year. In this regard, a shift in serotype predominance from G1 in 2000-2001 (61.8%) and 2001-2002 (69.7%) to G9 in 2002-2003 (51.3%) was an intriguing observation that has been reported recently in some other countries, as well. The emergence of serotype G9 rotaviruses in Hungary and other parts of the world may have implications for future vaccine development and use, particularly, if current vaccine candidates cannot confer adequate homotypic or heterotypic protection against these strains.

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“…For PAGE, dsRNA was extracted from the infected CrFK cells using the guanidine thiocyanate/Glass Milk method (Gentsch et al, 1992), run in a 10% polyacrylamide gel at constant current of 10 mA for 10 h and visualized by silver staining as previously described (Dolan et al, 1985).…”

Section: Polyacrylamide Gel Electrophoresis (Page)mentioning

confidence: 99%

Virological and molecular characterization of a mammalian orthoreovirus type 3 strain isolated from a dog in Italy

Decaro¹,

Campolo²,

Desario³

et al. 2005

Veterinary Microbiology

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A mammalian orthoreovirus (MRV) strain was isolated from a pup with fatal diarrhea, which had a concurrent infection by canine parvovirus type 2. The reovirus isolate showed an atypical hemagglutination pattern and a retarded electrophoretic mobility of the S1 segment, which is characteristic of MRV type 3 (MRV-3). Assignment of the isolated virus to MRV-3 was confirmed by type-specific RT-PCR assays, targeting the S1 gene, and by subsequent sequence analysis of the PCR product. By phylogeny based on the S1 gene of several MRVs, the isolate fell into lineage E, along with the murine strain T3C9/61 and the bovine strains T3C18/61 and T3C31/59. Conversely, L1 sequences were found to segregate regardless of the viral type. A total of 110 fecal samples, 56 nasal and 31 ocular swabs from dogs with diarrhea or nasal/ocular discharge were tested by a nested-PCR assay specific for reoviruses, and no sample was found to contain MRV RNA, a finding that is apparently in contrast with the seroprevalence (25.77%) observed in dogs.

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Epidemiology of rotavirus electropherotypes determined by a simplified diagnostic technique with RNA analysis

Cited by 73 publications

References 32 publications

Chronic enteric virus infection in two T‐cell immunodeficient children

Chronic enteric virus infection in two T‐cell immunodeficient children

Dominating prevalence of P[8],G1 and P[8],G9 rotavirus strains among children admitted to hospital between 2000 and 2003 in Budapest, Hungary

Virological and molecular characterization of a mammalian orthoreovirus type 3 strain isolated from a dog in Italy

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