1887

Abstract

SUMMARY

Two monoclonal antibodies (M-Ab) specific for different epitopes on particles of soybean mosaic virus (SMV) were used in a double-antibody sandwich ELISA (M-Ab ELISA). The non-isotopic immunoassay, which used a biotinylated second antibody and an avidin-alkaline phosphatase detection system to detect SMV in soybean seed extracts, was compared with a polyclonal antibody-based solid-phase radioimmunoassay (SPRIA). -Ab ELISA detected less than 10 ng of SMV/ml and was more sensitive than the SPRIA which detected 25 ng SMV/ml. Furthermore, -Ab ELISA required less than 36 h for seed sample assays and only 5.5 h for assays involving purified virus, whereas SPRIA required 3 or 4 days. When seeds from 33 field plots, in which 0%, 30% and 50% of the soybean plants had been inoculated with SMV, were assayed by both systems, results of the two tests correlated for 31 of 33 (94%) seed samples. This suggests that dual-site biotin-avidin -Ab ELISA systems have potential utility for routine screening of seed samples.

Keyword(s): biotin-avidin , ELISA , seeds and SMV
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/content/journal/jgv/10.1099/0022-1317-66-10-2089
1985-10-01
2024-04-25
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References

  1. Abney T. S., Sillings J. O., Richards T. L., Broersma D. B. 1976; Aphids and other insects as vectors of soybean mosaic virus. Journal of Economic Entomology 69:254–256
    [Google Scholar]
  2. Bayer E. A., Skutelsky E., Wilchek M. 1979; The avidin-biotin complex in affinity cytochemistry. Methods in Enzymology 62:308–326
    [Google Scholar]
  3. Bowers G. R., Goodman R. M. 1979; Soybean mosaic virus: infection of soybean seed parts and seed transmission. Phytopathology 60:569–572
    [Google Scholar]
  4. Briand J. P., Al Moudallal Z., Van Regenmortel M. H. V. 1982; Serological differentiation of tobamoviruses by means of monoclonal antibodies. Journal of Virological Methods 5:293–300
    [Google Scholar]
  5. Bryant G. R., Hill J. H., Bailey T. B., Tachibana H., Durand D. P., Benner H. I. 1982; Detection of Soybean mosaic virus in seed by solid-phase radioimmunoassay. Plant Disease 66:693–695
    [Google Scholar]
  6. Bryant G. R., Durand D. P., Hill J. H. 1983; Development of a solid-phase radioimmunoassay for detection of soybean mosaic virus. Phytopathology 73:623–629
    [Google Scholar]
  7. Chen L. -C., Durand D. P., Hill J. H. 1982; Detection of soybean mosaic virus pathogenic strains by enzyme-immunosorbent assay using polystyrene plates and beads as the solid phase. Phytopathology 72:1177–1181
    [Google Scholar]
  8. Diaco R., Lister R. M., Durand D. P., Hill J. H. 1983; Production of monoclonal antibodies against three isolates of barley yellow dwarf virus. Phytopathology 73:788
    [Google Scholar]
  9. Diaco R., Hill J. H., Lister R. M., Durand D. P. 1984; Use of monoclonal antibodies in serologically specific j electron microscopy of barley yellow dwarf virus. Phytopathology 74:860
    [Google Scholar]
  10. Fehr W. R., Caviness D. E. 1977; Stages of soybean development. Special Report. Iowa Cooperative Extension Service80
    [Google Scholar]
  11. Guesdon J. -L., Ternyck T., Avrameas S. 1979; The use of avidin-biotin interaction in immunoenzymatic techniques. Journal of Histochemistry and Cytochemistry 27:1131–1139
    [Google Scholar]
  12. Gugerli P., Fries P. 1983; Characterization of monoclonal antibodies to potato virus Y and their use for virus detection. Journal of General Virology 64:2471–2477
    [Google Scholar]
  13. Halk E. L., Hsu H. T., Aebig J., Franke J. 1984; Production of monoclonal antibodies against three ilarviruses and alfalfa mosaic virus and their use in serotyping. Phytopathology 74:367–372
    [Google Scholar]
  14. Hill E. K., Hill J. H., Durand D. P. 1984; Production of monoclonal antibodies to viruses in the potyvirus group: use in radioimmunoassay. Journal of General Virology 65:525–532
    [Google Scholar]
  15. Hill J. H., Benner H. I. 1980; Properties of soybean mosaic virus and its isolated protein. Phytopathologische Zeitschrift 97:272–281
    [Google Scholar]
  16. Hill J. H., Bryant G. R., Durand D. P. 1981; Detection of plant virus by using purified IgG in ELISA. Journal of Virological Methods 3:27–35
    [Google Scholar]
  17. Hoffmann K., Finn F. M., Kiso Y. 1978; Avidin-biotin affinity columns. General methods for attaching biotin to peptides and proteins. Journal of the American Chemical Society 100:3585–3590
    [Google Scholar]
  18. Hsu H. T., Aebig J., Rochow W. F. 1984; Differences among monoclonal antibodies to barley yellow dwarf virus. Phytopathology 75:600–605
    [Google Scholar]
  19. Koenig R. 1978; ELISA in the study of homologous and heterologous reactions of plant virus. Journal of General Virology 40:309–318
    [Google Scholar]
  20. Köhler G., Milstein C. 1975; Continuous cultures of fused cells secreting antibody of predefined specificity. Nature, London 256:495–497
    [Google Scholar]
  21. Lister R. M. 1978; Application of enzyme-linked immunosorbent assay for detecting viruses in soybean seeds and plants. Phytopathology 68:1393–1400
    [Google Scholar]
  22. Lucas B. S., Hill J. H. 1980; Characteristics of the transmission of three soybean mosaic virus isolates by Myzus persicae and Rhopalosiphum maidis. Phytopathologische Zeitschrift 99:47–53
    [Google Scholar]
  23. McLaughlin M. R., Bryant G. R., Hill J. H., Benner H. I., Durand D. P. 1980; Isolation of specific antibody to plant viruses by acid sucrose density gradient centrifugation. Phytopathology 70:831–834
    [Google Scholar]
  24. Purcifull D. E. 1966; Some properties of tobacco etch virus and its alkaline degradation products. Virology 29:8–14
    [Google Scholar]
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