A sensitive, single-radial diffusion autoradiographic zone size enhancement technique for the assay of influenza haemagglutinin

J Gen Virol. 1980 Apr;47(2):355-63. doi: 10.1099/0022-1317-47-2-355.

Abstract

A novel radial-diffusion technique in agarose gels is described which is applicable to the assay of small concentrations of influenza haemagglutinin (HA) antigen. The method is based on single-radial diffusion zone size enhancement (ZE) using autoradiography to demonstrate antigen--antibody reactions. The ZE technique is capable of reliably assaying concentrations of haemagglutinin as low as 0.1 microgram/ml, which represents a 20- to 40-fold increase in sensitivity over conventional single-radial-diffusion techniques. The ZE response is dependent upon the antigenic identity of the 125I-labelled 'marker' HA and the HA antigen being assayed. Haemagglutinins showing variation within a subtype (e.g. H3) show reduced ZE responses and thus the method has potential value for distinguishing between closely related HA antigens.

MeSH terms

  • Autoradiography
  • Hemagglutinins, Viral / analysis*
  • Hemagglutinins, Viral / classification
  • Immunodiffusion / methods*
  • Influenza A virus / immunology*
  • Orthomyxoviridae / immunology

Substances

  • Hemagglutinins, Viral