Detection of antibodies to rabbit haemorrhagic disease virus: an immunoblotting method using virus-coated human erythrocyte membranes.

The virus of rabbit haemorrhagic disease (RHDV) was purified from infected rabbit liver homogenate by using its property to bind to human red blood cells. Lysates from virus coated cells contained a 60 kDa protein identified as the major viral protein. Immunoblots prepared with that preparation were proved to be useful for immunochemical analysis since the 60 kDa component was intensively stained by subsequent incubation with rabbit sera from infected rabbits and with a secondary labelled antibody. The sera from 114 rabbits were analysed with this test and the data were compared with those obtained by using the haemagglutination inhibition test (HIT). Among the 114 field sera tested by Western blot, 86 contained antibodies to the 60 kDa RHDV antigen whereas only 76 showed positive reaction by HIT. The sensitivity and the specificity of the Western blot were 0.85 and 0.45, respectively, with a concordance between the two techniques of 0.72. Additionally, the European brown hare syndrome virus antibodies reacted with the 60 kDa RHDV protein on immunoblots.