Immunoassay of Poliovirus Antigens by Single-Radial-Diffusion: Development and Characteristics of a Sensitive Autoradiographic Zone Size Enhancement (ZE) Technique Free

Abstract

SUMMARY

The reactions of polioviruses in single-radial-immunodiffusion (SRD) tests were investigated with a view to developing accurate and sensitive antigen assay systems. In direct SRD tests, employing high concentrations of immune poliovirus serum in agarose gels, poliovirus D-antigens produced clear reaction zones demonstrated by protein staining. The reactions were type-specific for polioviruses of types 1, 2 and 3 but the tests were of low sensitivity, being applicable only to the assay of virus concentrates.

A novel autoradiographic zone size enhancement (ZE) test was developed which increased the sensitivity of the SRD assay 40- to 100-fold. The ZE test was dependent upon the ability of unlabelled poliovirus to co-migrate with the radioactive marker virus and so enhance the zone size detected autoradiographically. The areas of the autoradiographic zones were directly proportional to the concentration of unlabelled antigen. The ZE test was capable of detecting poliovirus D antigens in diluted cell culture fluid harvests in amounts corresponding to 10 to 10 TCID of infectious virus.

Studies with poliovirus type 3 strains indicated that the ZE test was narrowly strain-specific for the D-antigen of poliovirus type 3 strains when homologous type 3 D-antigen was used as radioactive marker, but broadly cross-reactive for the D-antigen of type 3 viruses when heterologous poliovirus type 3 D-antigen was used as marker.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-51-1-157
1980-11-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/51/1/JV0510010157.html?itemId=/content/journal/jgv/10.1099/0022-1317-51-1-157&mimeType=html&fmt=ahah

References

  1. Beale A. J., Mason P. J. 1962; The measurement of the D-antigen in poliovirus preparations. Journal of Hygiene 60:113–120
    [Google Scholar]
  2. Cowan K. M., Wagner G. G. 1970; Immunochemical studies of foot and mouth disease. VIII. Detection and quantitation of antibodies by radial immunodiffusion. Journal of Immunology 105:557–566
    [Google Scholar]
  3. Dömök I., Magrath D. I. 1979; Poliovirus isolation and serological techniques. World Health Organization publication no. 46
    [Google Scholar]
  4. Esposito J. J. 1976; Detection of poliovirus antigens and antibodies: micro-indirect haemagglutination and haemagglutination inhibition tests for poliovirus types 1, 2 and 3. Microbios 16:29–36
    [Google Scholar]
  5. Gard S., Wesslen T., Fagraeus A., Svedmyr A., Olin G. 1956; The use of guinea pigs in tests for immunogenic capacity of poliomyelitis virus preparations. Archiv für die Gesamte Virusforschung 6:401–411
    [Google Scholar]
  6. Ghendon Y. Z., Yakobson E. A. 1971; Antigenic specificity of poliovirus-related particles. Journal of Virology 8:589–590
    [Google Scholar]
  7. Le Bouvier G. L. 1955; The modification of poliovirus antigens by heat and ultraviolet light. Lancet ii:1013–1016
    [Google Scholar]
  8. Le Bouvier G. L. 1959; Poliovirus D and C antigens–their differentiation and measurement by precipitin in agar. British Journal of Experimental Pathology 40:452–460
    [Google Scholar]
  9. Mancini G., Carbonara O. A., Heremans J. F. 1965; Immunochemical quantitation of antigens by single-radial-immunodiffusion. Immunochemistry 2:235–254
    [Google Scholar]
  10. Mayer M. M., Rapp H. J., Roizman B., Klein S. W., Cowan K. M., Lukens D., Schwerdt C. E., Schaffer F. L., Charney J. 1957; The purification of poliomyelitis virus as studied by complement fixation. Journal of Immunology 78:435–455
    [Google Scholar]
  11. Minor P. D. 1980; Comparative biochemical studies of type 3 poliovirus. Journal of Virology 34:73–84
    [Google Scholar]
  12. Minor P. D., Schild G. C., Wood J. M., Dandawate C. N. 1980; The preparation of specific immune sera against type 3 poliovirus D-antigen and C-antigen and the demonstration of two C-antigen components in vaccine strain populations. Journal of General Virology 51:147–156
    [Google Scholar]
  13. Pereira H. G., Machado R. D., Schild G. C. 1972; Study of adenovirus hexon antigen-antibody reactions by single-radial-diffusion techniques. Journal of Immunological Methods 2:121–128
    [Google Scholar]
  14. Roizman B., Hopken W., Mayer M. M. 1957; Immunochemical studies of poliovirus. II. Kinetics of formation of infectious and non-infectious type 1 poliovirus in three cell strains of human derivation. Journal of Immunology 80:386–395
    [Google Scholar]
  15. Roizman B., Mayer M. M., Rapp H. 1958; Immunochemical studies of polioviruses. VI. Further studies on the immunologic and physical properties of poliovirus particles produced in cell culture. Journal of Immunology 81:419–425
    [Google Scholar]
  16. Rumke Ph., Thung P. J. 1964; Immunological studies on the sex-dependent prealbumin in mouse urine and its occurrence in the serum. Acta Endocrinologica 47:156–164
    [Google Scholar]
  17. Schild G. C., Wood J. M., Newman R. W. 1975; A single-radial-immunodiffusion technique for the assay of influenza haemagglutinin antigen. Bulletin of the World Health Organization 52:223–230
    [Google Scholar]
  18. Van Wezel A. L., Hazendonk A. G. 1979; Intratypic serodifferentiation of poliomyelitis virus strains by specific antisera. Intervirology 11:2–8
    [Google Scholar]
  19. Wood J. M., Schild G. C., Newman R. W., Seagroatt V. 1977; A single-radial-immunodiffusion technique for the assay of influenza haemagglutinin. Bulletin of the World Health Organization 52:223–230
    [Google Scholar]
  20. Wood J. M., Schild G. C., Newman R. W. 1980; A sensitive, single-radial-diffusion autoradiographic zone size enhancement technique for the assay of influenza haemagglutinin. Journal of General Virology 47:355–363
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-51-1-157
Loading
/content/journal/jgv/10.1099/0022-1317-51-1-157
Loading

Data & Media loading...

Most cited Most Cited RSS feed