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 to 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.
Czerny
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Institute for Medical Microbiology, Infectious and Epidemic Diseases, Ludwig-Maximilians-University, Munich, Germany.
Wagner
K
K
Gessler
K
K
Mayr
A
A
Kaaden
O R
OR
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000097:blood* / Q000276:immunology; D000918:Antibody Specificity; D001667:Binding, Competitive; D015153:Blotting, Western; D002371:Cat Diseases / Q000453:epidemiology*; D002415:Cats; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary*; D005858:Germany / Q000453:epidemiology; D006825:Hybridomas; D051379:Mice; D009500:Neutralization Tests / Q000662:veterinary; D018155:Orthopoxvirus / Q000276:immunology*; D011213:Poxviridae Infections / Q000453:epidemiology / Q000662:veterinary*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D016036:Seroepidemiologic Studies
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral