Detection of Actinobacillus pleuropneumoniae serotype 2 antibodies in pig sera by an inhibition enzyme immuno assay (EIA).
An inhibition Enzyme Immuno Assay (EIA) for detection of antibodies against A. pleuropneumoniae serotype 2 (App-2) in pig sera, based on the inhibition of the binding of an App-2 specific monoclonal antibody was established. The monoclonal antibody (mAb 102-G02) was found to be directed against an epitope on the O-chain of App-2 LPS. Some App-2 isolates did not react with the mAb 102-G02. These isolates are referred to as App-2X. In the inhibition EIA highly purified App-2 LPS ws used to coat microtiter plates. Serial dilutions of pig sera were added to the plates prior to the mAb 102-G02. The degree of binding of App-2 antibodies from pig sera was determined as the percentage inhibition of the mAb 102-G02. Pig sera from specific pathogen free (SPF) herds, from experimentally infected animals, and from conventionel herds were tested. A serum dilution of 1/200 was found to be optimal, when using 50% inhibition as the discriminating inhibition percentage. Serum from App-2X infected herds showed a lower reactivity as compared to serum from App-2 infected herds. No crossreactivity was observed with serum from pigs infected with other App serotypes. The sensitivity and specificity were 100% and 98.9%, respectively.
Stenbaek
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EI
National Veterinary Laboratory, Copenhagen, Denmark.
Schirmer
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D000189:Actinobacillus Infections / Q000175:diagnosis / Q000662:veterinary*; D016977:Actinobacillus pleuropneumoniae / Q000276:immunology*; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology; D003168:Complement Fixation Tests; D003429:Cross Reactions; D007106:Immune Sera / Q000276:immunology; D007124:Immunoenzyme Techniques / Q000662:veterinary; D051379:Mice; D011001:Pleuropneumonia / Q000175:diagnosis / Q000662:veterinary*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013047:Specific Pathogen-Free Organisms; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis*
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D007106:Immune Sera