Abstract
The results of RNA analysis for the detection of rotavirus were compared with those of a standard enzyme-linked immunsorbent assay (ELISA) and electron microscopy using 212 faecal specimens obtained from 200 children with gastroenteritis. Rotavirus was extracted directly from faecal specimens and RNA segments were made visible by polyacrylamide gel electrophoresis using a silver staining technique. Of the 212 faecal specimens 137 were found to be positive in ELISA, 125 in RNA analysis and 121 in both methods. Forty-nine of the 212 specimens were also investigated by electron microscopy. Thirty-five were positive when examined by electron microscopy, 37 were positive in ELISA and 33 in RNA analysis. RNA analysis of 119 faecal samples in outbreaks and sporadic cases of rotavirus infection yielded 42 different rotavirus electrophoretypes. The results indicated that no one method was sufficient to detect all positive specimens and that RNA analysis is useful in epidemiological studies.
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Wyatt, R. G., Kalica, A. R., Mebus, C. A., Kim, H. W., London, W. T., Chanock, R. M., Kapikian, A. Z.: Reovoris-like agents (rotaviruscs) associated with diarrheal illness in animals and man. Perspectives in Virology 1978, 10: 121–145.
Holmes, J. H., Ruck, B. J., Bishop, R. F., Davidson, G. P.: Infantile enteritis viruses: morphogenesis and morphology. Journal of Virology 1975, 16: 937–943.
Baumeister, H. G., Balks, H. G., Maass, G.: Elektronenmikroskopischer Direktnachweis von Viruspartikel im Säuglings und Kleinkindalter. Klinische Wochenschrift 1976, 54: 445–558.
Yolken, R. H., Barbour, B., Wyatt, R. G., Kalica, A. R., Kapikian, A. Z., Chanock, R. M.: Enzyme-linked immunosorbent assay for identification of rotaviruses from different animal species. Science 1978, 201: 259–261.
Yolken, R. H., Wyatt, R. G., Zissis, G., Brandt, C. D., Rodriguez, W. J., Kim, H. W., Parrott, R. H., Urrutia, J. J., Mata, L., Greenberg, H. B., Kapikian, A. Z., Chanock, R. M.: Epidemiology of human rotavirus types 1 and 2 as studied by enzyme-linked immunosorbent assay. New England Journal of Medicine 1978, 299: 1156–1161.
Kalica, A. R., Greenberg, H. B., Espejo, R. T., Flore, J., Wyatt, R. G., Kapikian, A. Z., Chanock, R. M.: Distinctive ribonucleic acid patterns of human rotavirus subgroups 1 and 2. Infection and Immunity 1981, 33: 958–961.
Thouless, M. E., Beards, G. M., Flewett, T. H.: Serotyping and Subgrouping of rotavirus strains by the ELISA test. Archives of Virology 1982, 73: 219–230.
Zissis, G., Lambert, J. P.: Enzyme-linked immunosorbent assays adapted for serotyping of human rotavirus strains. Journal of Clinical Microbiology 1980, 11: 1–5.
Kalica, A. R., Garon, C. F., Wyatt, R. G., Mebus, C. A., van Kirk, D. H., Chanock, R. M., Kapikian, A. Z.: Differentiation of human and calf reoviruslike agents associated with diarrhea using polyacrylamide gel electrophoresis of RNA.. Virology 1976, 74: 86–92.
Rodger, S. M., Holmes, I. H.: Comparison of the genomes of simian, bovine, and human rotaviruses by gel electrophoresis and detection of genomic variation among bovine isolates. Journal of Virology 1979, 30: 839–846.
Sabara, M. M., Deregt, D., Babiuk, L. A., Misra, V.: Genetic heterogeneity within individual bovine rotavirus isolates. Journal of Virology 1982, 44: 813–822.
Herring, A. J., Inglis, N. F., Ojeh, C. K., Snodgrass, D. R., Menzies, J. D.: Rapid diagnosis of rotavirus infection by direct detection of viral nucleic acid in silber-stained polyacrylamide gels. Journal of Clinical Microciology 1982, 16: 473–477.
Theil, W, K., McCloskey, C. M., Saif, L. J., Redman, D. R., Bohl, E. H., Hancock, D. D., Kohler, E. M., Moorhead, P. D.: Rapid and simple method of preparing rotaviral double-stranded ribonucleic acid for analysis by polyacrylamide gel electrophoresis. Journal of Clinical Microbiology 1981, 14: 273–280.
Kalica, A. R., Sereno, M. M., Wyatt, R. G., Mebus, C. A., Chanock, R. M., Kapikian, A. Z.: Comparison of human and animal rotavirus strains by gel electrophoresis of viral RNA. Virology 1978, 87: 247–255.
Spencer, E. G., Avendano, L. F., Garcia, B. I.: Analysis of human rotavirus mixed electropherotypes. Infection and Immunity 1983, 39: 569–574.
Spencer, E. G., Avedano, F., Araya, M.: Characteristics and analysis of electropherotypes of human rotavirus isolated in Chile. Journal of Infectious Diseases 1983, 148: 41–48.
Espejo, R. T., Munoz, O., Serafin, F., Romero, R.: Shift in the prevalent human rotavirus detected by ribonucleic acid segment differences. Infection and Immunity 1980, 27: 351–354.
Estes, M. K., Graham, D. Y., Dimitrov, D. H.: The molecular epidemiology of rotavirus gastroenteritis. Progress in Medical Virology 1984, 29: 1–22.
Narang, H. K., Codd, A. A.: Preponderance of rotavirus in clumped form in patients with acute gastroenteritis. Lancet 1980, i: 1192–1193.
Narang, H. K., Codd, A. A.: Frequency of preclumped virus in routine fecal specimens from patients with acute nonbacterial gastroenteritis. Journal of Clinical Microbiology 1981, 13: 982–988.
Kirby, K. S.: Isolation of nucleic acids with phenoloc solvents. In: Colowick, S. P., Kaplan, N. O. (ed.): Methods in enzymology. Academic Press, New York, 1968, p. 87–98.
Laemmli, U. K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970, 227: 680–685.
Merril, C. R., Dunau, M. L., Goldman, D.: A parid sensitive stain for polypeptides in polyacrylamide gels. Analytical Biochemistry 1981, 110: 201–207
Gelderblom, H., Reupke, H., Warring, R.: Über den Einsatz der Airfuge in der elektronenmikroskopischen Virusdiagnostik. Fachzeitschrift für das Laboratorium 1978, 1: 17–19.
Follett, E. A. C., Desselberger, U.: Co-circulation of different rotavirus strains in a local outbreak of infantile gastroenteritis: monitoring by rapid and sensitive nucleic acid analysis. Journal of Medical Virology 1983, 11: 39–52.
Brandt, C. D., Kim, H. W., Rodriguez, W. J., Thomas, L., Yolken, R. H., Arrobio, J. O., Kapikian, A. Z., Parrott, R. H., Channock, R. M.: Comparison of direct electron microscopy and rotavirus enzyme-linked immunosorbent assay for detection of gastroenteritis viruses in children. Journal of Clinical Microbiology 1981, 13: 976–981.
Cheung, E. Y., Hnatko, S. I., Gunning, H., Wilson, J.: Comparison of Rotazyme and direct electron microscopy for detection of rotavirus in human stools. Journal of Clinical Microbiology 1982, 16: 562–653.
Chasey, D., Labram, J.: Electron microscopy of tubular assemblies associated with naturally occuring bovine rotavirus. Journal of General Virology 1983, 64: 863–872.
Follett, E. A. C., Sanders, R. C., Beards, G. M., Hundley, F., Desselberger, U.: Molecular epidemiology of human rotaviruses. Analysis of outbreaks of acute gastroenteritis in Glasgow and the west of Scotland 1981/82 and 1982/83. Journal of Hygiene 1984, 92: 209–222.
Young, J. F., Palese, P.: Evolution of human influenza A viruses in nature: recombination contributes to genetic variation of H1N1 strains. Proceedings of National Academy of Sciences of the United States of America 1979, 76: 6547–6551.
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Selb, B., Baumeister, H.G., Maass, G. et al. Detection of human rotavirus by nucleic acid analysis in comparison to enzyme-linked immunoassay and electron microscopy. Eur. J, Clin. Microbiol. 4, 41–45 (1985). https://doi.org/10.1007/BF02148658
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DOI: https://doi.org/10.1007/BF02148658