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 µg/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 I-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.


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