Studies On A Dot-PPA-ELISA Method For The Detection Of Antibody Against Pasteurellosis

A Dot-PPA-ELISA method has been developed to detect the antibody in swine serum against pasteurellosis. The capsule polysaccharide and outer membrane protein of pasteurella multocida was prepared as the diaphragm antigen. Applying the x2 test,optimal concentration of the diaphragm antigen was determined,and the optimum dilution of enzyme HRP-SPA 1:10. The high specificity of the Dot-PPA-ELISA was confirmed by specific blocking test and also by cross-reaction test. The diaphragm did not react with the antibodies against Salmonellosis,Streptococcosis,Colibacillosis,Chlamydiosis,HCV,PPV,PRV,Brucellosis. Erysipelas,suis and Chlamydiosis in cross-reaction test.The diaphragm has good sensitivity and could detect some pasteurella-positive test serum which has been diluted to2-". In comprision with GDPT ,the sensetivity is improved by 128 times . When 10 positive serum was detected 3 times,and the result was reproducible. The diaphragm has good stability as well,stored at 4 for at least 5 months or at 10-25 for 3 months,the sensitivity and specificity of the diaphragm did not change.This study endicates that the Dot-PPA-ELISA was a convenient,rapid,sensitive and specific method for the detection of antibody. Apart from these,all of the materials used in Dot-PPA-ELISA can be standardized. So it can be used for the surveillance of pasteurella multocida antibody in herd ,for the epidemiologic investigation of swine pasteurellosis ,and it provides a reference for a ratinal immunity schedule.