DETECTION OF ANTIBODIES TO RABBIT HEMORRHAGIC-DISEASE VIRUS - AN IMMUNOBLOTTING METHOD USING VIRUS-COATED HUMAN ERYTHROCYTE-MEMBRANES

The virus of rabbit haemorrhagic disease (RHDV) was purified from infe cted rabbit liver homogenate by using its property to bind to human re d blood cells. Lysates from virus coated cells contained a 60 kDa prot ein identified as the major viral protein, Immunoblots prepared with t hat preparation were proved to be useful for immunochemical analysis s ince the 60 kDa component was intensively stained by subsequent incuba tion with rabbit sera from infected rabbits and with a secondary label led antibody. The sera from 114 rabbits were analysed with this test a nd the data were compared with those obtained by using the haemaggluti nation inhibition test (HIT). Among the 114 field sera tested by Weste rn blot, 86 contained antibodies to the 60 kDa RHDV antigen whereas on ly 76 showed positive reaction by HIT. The sensitivity and the specifi city of the Western blot were 0.85 and 0.45, respectively, with a conc ordance between the two techniques of 0.72. Additionally, the European brown hare syndrome virus antibodies reacted with the 60 kDa RHDV pro tein on immunoblots.