A monoclonal blocking-ELISA for detection of orthopoxvirus antibodies in feline sera

A double sandwich blocking-ELISA using a genus-specific neutralizing monoclonal antibody (MAb) against the vaccinia virus 32 kD adsorption protein (D8L open reading frame; ORF) was developed to detect orthopoxvirus (OPV) antibodies in sera. A collection of 2173 feline serum samples was examined in an epidemiological study. The blocking-ELISA revealed 44 (2%) sera with positive titres of 1:2-1:256. ELISA results were confirmed by the plaque-reduction test. A close correlation between titres of both assays could be observed ( r = 0.986). In general, the sensitivity of the blocking ELISA was two to four times higher. Neutralizing OPV-antibodies were found in nine sera with ELISA-titres > 1:4. Antibody specificity of OPV was also demonstrated by Western blotting analysis with selected feline sera. The epidemiographical distribution of the ELISA-positive sera and case histories of 37 seropositive cats available from the referring veterinarians are demonstrated. The blocking-ELISA enables a rapid serological diagnosis and can be used in veterinary and human medicine. It allows OPV-antibody screening in human and other animal species.