1887

Abstract

Summary

A time-resolved fluorimmunoassay (TRFIA) with europium-labelled monoclonal antibodies (MAbs) was used to detect potato virus X (PVX) at a concentration of 5 pg/ml, in potato tuber extracts diluted up to 7 × 10-fold and leaf extracts diluted up to 2 × 10-fold. When mixed with potato leaf sap, PVX was detected at 100 pg/ml. TRFIA with simultaneous incubation of antigen and labelled antibody (one incubation) was two orders of magnitude more sensitive for PVX detection than the conventional double antibody sandwich ELISA. The fluorescence signal in TRFIA with separate incubations of antigen and labelled antibody (two incubations) with MAbs was linearly related to the virus concentration between 1.6 ng/ml and 1000 ng/ml; the method can therefore be used for the quantification of PVX. PVX detection in plant specimens by one incubation ELISA with MAbs was generally 10 times more sensitive than the standard two incubation procedure. Europium TRFIA with MAbs to potato viruses was found to be a very sensitive method for the detection of PVX, potato virus M, potato virus S, potato virus Y and potato leaf roll virus.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-69-5-991
1988-05-01
2024-05-03
Loading full text...

Full text loading...

/deliver/fulltext/jgv/69/5/JV0690050991.html?itemId=/content/journal/jgv/10.1099/0022-1317-69-5-991&mimeType=html&fmt=ahah

References

  1. BOBKOVA A. F., NACVLISHVILI N. M., SOROKINA N. B., EGOROV A. M., ATABEKOV I. G. 1983; Application of immunoferment method to diagnosis of potato X-virus diseases in potato plants and tubers. Sel’sko-hozyaistvennaya biologiya 18:115–118 in Russian
    [Google Scholar]
  2. CLARK M. F., ADAMS A. N. 1977; Characteristics of the microplate method of enzyme-linked immunosorbent assay for the detection of plant viruses. Journal of General Virology 34:475–483
    [Google Scholar]
  3. CLARK M. F., LISTER R. M., BAR-JOSEPH M. 1986; ELISA technique. Methods in Enzymology 118:742–766
    [Google Scholar]
  4. EKINS R. P. 1985 Current concepts and future developments. Alternative Immunoassays219–237 Edited by Collins W. P. Chichester & New York: John Wiley & Sons;
    [Google Scholar]
  5. ENGVALL E., PERLMANN P. 1971; Enzyme-linked immunosorbent assay (ELISA). Quantitative assay of IgG. Immunochemistry 8:871–874
    [Google Scholar]
  6. FLEGG C. L., CLARK M. F. 1979; The detection of apple chlorotic leafspot virus by a modified procedure of enzyme-linked immunosorbent assay (ELISA). Annals of Applied Biology 91:61–65
    [Google Scholar]
  7. HALONEN P., BONFANTI C, LÖVGREN T., HEMMILä I., SOINI E. 1985 Detection of viral antigens by time-resolved fluoroimmunoassay. Rapid Methods and Automation in Microbiology and Immunology429–437 Edited by Habermehl K.-O. Heidelberg: Springer-Verlag;
    [Google Scholar]
  8. 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]
  9. HALK E. L., DE BOER S. H. 1985; Monoclonal antibodies in plant disease research. Annual Review of Phytopathology 23:321–350
    [Google Scholar]
  10. HEMMILÄ I. 1985; Fluoroimmunoassays and immunofluorometric assays. Clinical Chemistry 31:359–370
    [Google Scholar]
  11. HEMMILä I., DAKUBU S., MUKKALA V.-M., SIITARI H., LöVGREN T. 1984; Europium as a label in time-resolved immunofluorometric assays. Analytical Biochemistry 137:335–343
    [Google Scholar]
  12. HSU H. 1982 Plant viruses and antisera. ATCC Catalog of Strains I545–571 Edited by Daggett P.-M., Gherna R. L., Pienta P., Nierman W., Jong S.-C., Hsu H., Brandon B., Alexander M. T. Rockville: American Type Culture Collection;
    [Google Scholar]
  13. JäRVEKüLG L., SOBER J., SINUÄRV R., SAARMA M. 1987; Use of time-resolved fluoroimmunoassay for testing of plant viruses. Doklady VASHNIL 12:15–18 in Russian
    [Google Scholar]
  14. LAEMMLI U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  15. LÖVGREN T., HEMMILä I., PETTERSON K., HALONEN P. 1985 Time-resolved fluorometry in immunoassay. Alternative Immunoassays203–217 Edited by Collins W. P. Chichester & New York: John Wiley & Sons;
    [Google Scholar]
  16. RADAVSKY I. L., GRAMA D. L., ZAIKIN A. A., WITER S. S., TUROVA I. P., BOBKOVA A. F., GOLDSTEIN M. I., JÄRVEKÜLG L. V., SOBER J. P., SAARMA M. J. 1988; The antigenic structure of potato virus X coat protein. I. The immunochemical analysis of cyanogen bromide fragments. Bioorganicheskaya khimiya 14:20–26 in Russian
    [Google Scholar]
  17. SHALLA T. A., SHEPARD J. F. 1970; An antigenic analysis of potato virus X and its degraded protein. II. Evidence for a conformational change associated with the depolymerization of structure protein. Virology 42:835–847
    [Google Scholar]
  18. SIITARL H., KURPPA A. 1987; Time-resolved fluoroimmunoassay in the detection of plant viruses. Journal of General Virology 68:1423–1428
    [Google Scholar]
  19. SIITARI H., HEMMILä I., SOINI E., LöVGREN T., KOISTINEN V. 1983; Detection of hepatitis B surface antigen using time-resolved fluoroimmunoassay. Nature, London 301:258–260
    [Google Scholar]
  20. SOBER J., JÄRVEKÜLG L., TOOTS I., RADAVSKY J., VILLEMS R., SAARMA M. 1988; Antigenic characterization of potato virus X with monoclonal antibodies. Journal of General Virology in press
    [Google Scholar]
  21. SOINI E., KOJOLA H. 1983; Time-resolved fluorometer for lanthanide chelates - a new generation of nonisotopic immunoassays. Clinical Chemistry 29:65–68
    [Google Scholar]
  22. TORRANCE L. 1980; Use of bovine Clq to detect plant viruses in an enzyme-linked immunosorbent-type assay. Journal of General Virology 51:229–232
    [Google Scholar]
  23. TORRANCE L., LARKINS A. P., BUTCHER G. W. 1986; Characterization of monoclonal antibodies against potato virus X and comparison of serotypes with resistance groups. Journal of General Virology 67:57–67
    [Google Scholar]
  24. VAN WEEMEN B. K., SCHUURS A. H. W. M. 1971; Immunoassay using antigen-enzyme conjugates. FEBS tetters 15:232–236
    [Google Scholar]
  25. VAN REGENMORTEL M. H. V. 1986 The potential for using monoclonal antibodies in the detection of plant viruses. Developments and Applications in Virus Testing 189–101 Edited by Jones R. A. C., Torrance L. Wellesbourne: Lavenham Press;
    [Google Scholar]
  26. WILSON M. B., NAKANE P. K. 1978 Developments in the periodate method of conjugating horseradish peroxidase to antibodies. Immunofluorescence and Related Staining Techniques215–244 Edited by Knapp W., Holubar K., Wick G. Amsterdam & New York: Elsevier/North-Holland;
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-69-5-991
Loading
/content/journal/jgv/10.1099/0022-1317-69-5-991
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error