【作者】 李金海；

【导师】 文心田；

【作者基本信息】 四川农业大学 ， 预防兽医学， 2002， 硕士

【摘要】 本研究选取C、D、E、R群及2型链球菌标准菌株，以高压法提取抗原建立了检测猪链球菌病血清抗体的Dot-PPA-ELISA方法。试验中对多种方法提取的诊断抗原进行了比较、筛选；确定了Dot-PPA-ELISA的最佳工作条件；测定了ST-171株的免疫猪血清抗体及C群、2型人工感染猪血清抗体效价，并初步确定了该方法的阳性标准。该法以硝酸纤维素膜为固相载体，包被膜载抗原制成的诊断膜片具有良好的特异性：不与仔猪副伤寒、猪巴氏杆菌病、猪大肠杆菌病、猪衣原体病、猪瘟、猪细小病毒病、猪伪狂犬病、猪布氏杆菌、猪丹毒阳性血清反应；膜片具有良好的灵敏性，阳性血清作2^-11稀释亦能检出；膜片具有良好的稳定性，在4℃至少能保存7个月，其灵敏性不变。该方法操作简便快捷，2.5小时即可得出结果；结果客观，易于判定；重复性好；各种试剂材料均可做到标准化。该方法适合规模化养猪场链球菌病的免疫监测，疾病诊断，以及流行病学调查。更多还原

【Abstract】 A Dot-PPA-ELISA has been developed to detect the specific antibody against streptococcus suis .The antigen was isolated from group C D E R and type 2 streptococcus suis by three different method:(A)autoclave extraction method,(B) HCL-extraction method ,and(C) Fuller’s method. Experiment shows that the antigen extracted by autoclave is best for Dot-PPA-ELISA . Applying the x2 test ,optimal concentration of antigen was determined,and the optimum dilution of enzyme HRP-SPA was 1:10. The high specificity of Dot-PPA-ELISA was proved by the specific blocking test,and also by the cross-reaction test in which the diaphragm does not react with the antibodies against Salmonellosis,Pasteurellosis,Chlamydiosis,HCV,PPV,Brucellosis,Erysipelas suis,Colibacillosis,PRV. The diaphragm had the ability to detect the positive serum when it was diluted to 2’11 and so it has good sensitivity;Stored at 4 for at least 7 months,the sensitivity and specificity of the diaphragm did not change,so it has good stability ;When 10 positive serum was detected 3 times,the result is reproducible,so the diaphragm has good reproducible.Serums from experimental inoculated piglets was detected .The results showed that when the titer is l:16,the pigs were infected with streptococcus suis;and when 1:64,the pigs could survive after challege with streptococcus suis.All the results have shown that Dot-PPA-ELISA was a convenient,rapid,sensitive specific useful method for the detection of antibody. It could be used for the surveillance of herd immunity and for the examination of immunization quality .It could aslo be used for diagnosis and epidemiologic investigation of streptoccus suis.更多还原