中文摘要：

对布鲁氏菌黏附素SP41蛋白进行表达、纯化,以表达重组蛋白为包被抗原,建立了检测布鲁氏菌病绵羊血清抗体的间接ELISA方法。克隆布鲁氏菌SP41基因,PRC、酶切、测序鉴定正确后,构建pET-32a（＋）-SP41原核表达载体,转化表达菌E.coliBL21（DE3）,IPTG诱导表达重组SP41蛋白,纯化表达产物后包被酶标反应板,方阵滴定法进行最佳血清稀释度和最侍抗原浓度的筛选,结果显示重组蛋白SP41最佳抗原包被浓度为0.6mg/L,最佳血清稀释度为1：50。交叉试验、阻断试验、重复性试验表明,该方法重复性好、特异性强。利用此方法检测217份绵羊血清样本,结果表明该方法的阳性检出率要高于虎红平板试验和试管凝集试验。

英文摘要：

An indirect ELISA was developed for detecting serum antibody against Brucella in sheep,using the purified recombinant expression product of the SP41 gene as the coating antigen.The cloned SP41 gene,identified correctly by PCR,double enzyme and sequencing,was used to construct the prokaryotic expression vector pET-32a（＋）-SP41,and transformed into E.coli BL21 cells and induced by IPTG.The purified recombinant SP41 was coated on the ELISA plate for detecting the specific antibodies against Brucella,and the optimal condotions for coating antigen concentration and antisera dilution were 0.6 mg/L and 1：50.The crossing-test,reproductive test and blocking-test showed that the indirect ELISA was more speific and reproductive.217 lamb sera from the farms were examined by the indirect ELISA,and 29 positive sera were detected in all adult sheep sera,seropositivity rate was 13.36%.