单抗竞争ELISA检测猪沙门氏菌感染方法的建立与应用  

Establishment and Application of a MAb-based Competitive ELISA for Detection of Piglet's Paratyphoid

作　　者：殷月兰[1] 潘志明[1] 孟书霞[2] 黄金林[1] 焦新安[1] 

机构地区：[1]扬州大学生物科学与技术学院,江苏扬州225009 [2]扬州大学畜牧兽医学院,江苏扬州225009

出　　处：《中国预防兽医学报》2003年第1期65-68,76,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：高校优秀青年教师教学科研奖励计划资助;扬州大学生物科学与技术学院青年教师科研启动基金资助

摘　　要：以猪霍乱沙门氏菌作为包被抗原 ,利用沙门氏菌O7单克隆抗体酶结合物建立了竞争ELISA方法检测猪霍乱沙门氏菌抗体。经过研究确定抗原包被浓度为 2×1 0 8CFU/ml,血清检测稀释度为 1∶1 6,酶标抗体浓度为 1∶70 0 ,包被液为碳酸盐缓冲液 ( 0 .0 5MpH9.6)。应用单抗竞争ELISA和平板凝集试验 (PAT)同时对 5 0 6份血清进行猪霍乱抗体检测 ,PAT的检出率为 3 .60 % ,ELISA检出率为 5 .75 % ,两者符合率达 96.4%。试验结果表明 ,ELISA方法敏感性高 ,特异性强 ,稳定性和重复性好 ,操作简便。本方法的建立在猪群沙门氏菌感染的检疫、实验室诊断的标准化及流行病学调查方面具有应用价值。Optimum conditions of cELISA were developed as follows;2×10 8CFU/ml of killed Samlmonella choleraesuis in carbonate sodium buffer(0.05M,pH9.6)used to coat ELISA plated,1:700 dilution of HRP-labelled MAb O 7 as detecting regent,and 1:16 of sera as detecting samples.A total of 506 sera wre detected in parallel by both the cELISA and plate agglutination test(PAT).5.75% of them were detected positively as piglet's paratyphoid,while 3.60% of them wrer positive by PAT.These results showed that the cELISA has the advantage of higher sensitivity,specificity,reproducibility and stability.It would be very useful in diagnosis,surveillance of piglet's paratyphoid.

关 键 词：单抗竞争ELISA 应用 猪 霍乱沙门氏菌 检测方法 副伤寒 

分 类 号：S858.28[农业科学—临床兽医学]