摘要
应用ＥＤＴＡ处理和超声波裂解法提取多杀性巴氏杆菌的荚膜多糖和外膜蛋白抗原，包被酶标板，用ＨＲＰ－ＳＰＡ代替酶标抗体，建立了检测兔多杀性巴氏杆菌病抗体的间接法ＥＬＩＳＡ。试验确定抗原的包被浓度为６．０５μｇ／ｍｌ，ＨＲＰ－ＳＰＡ的适宜稀释度为１１００００，血清ＥＬＩＳＡ滴度在１４００以上判为阳性。测定了多杀性巴氏杆菌灭活菌苗免疫兔血清１０份，均呈阳性，抗体滴度在１１２８０～１４０９６０范围内呈正态分布；血清几何平均效价为１５１２０。重复４次测定１０份阳性血清，其ＯＤ值之标准差（ＳＤ）＜０．１０，变异系数（ＣＶ）＜１０．０％。本法与琼脂扩散沉淀试验相比，敏感度提高约１５７５倍。ＥＬＩＳＡ不仅敏感、特异、快速、简便，而且经济、易于重复，适于推广应用。
An enzyme-linked immunosorbent assay(ELISA) was developed to detect the rabbit antibody against Pasteurellosis. The capsule polysaccharide and outer membrane protein of pasteurella multocida was prepared as antigen by subjecting the bacterium to 10mM EDTA ultrasonicated, and collected the supernatant by centrifugation. The antigen was used to coat microdilution plates for binding rabbit antibody against Pasteurella multocida optimal concentration of 6.05μg/ml. Staphyloccocus protein A (SPA) was labelled with horserodish peroxidae (HRO) by the modified periodate sodium method. An ELISA titer of 1∶400 or greater was considered positive. Serum of ten rabbits immunized were examined and all gave positive results with a ELISA geometric mean tites 1∶5120. It was showed that the indirect ELISA was sensitive, spectific, rapid, simple, accurate and reproducible. The authors suggest that this method may be valuable for diagnosing Pasteurellosis.
出处
《中国兽药杂志》
北大核心
1997年第1期1-5,共5页
Chinese Journal of Veterinary Drug
关键词
多杀性巴氏杆菌
抗体
ELISA
检测
兔病
Pasteurella multocida , capsule polysaccharide and outer membrane protein of pasteurella multocida , antibody, ELSA, detection
