【摘要】 利用丝状霉形体山羊亚种标准株PG3制备抗原,经纯化后作为包被抗原建立了间接ELISA方法,用于检测丝状霉形体山羊亚种血清抗体。经方阵滴定确定最佳抗原包被浓度为1.09μg/mL,血清样品最佳稀释度为1∶64,兔抗羊IgG辣根过氧化物酶结合物的最佳稀释度为1∶40 000,抗原抗体最佳结合时间为1 h。判定标准为样品D490 nm值与标准阴性血清D490 nm值之比(P/N)≥3为阳性,≤2.5为阴性,介于二者之间的为可疑。重复性和稳定性试验证明,建立的间接ELISA的稳定性和重复性良好,与间接血凝试验相比,其敏感性较高。更多还原

【Abstract】 Antigen against Mycoplasma mycoides subsp.capri was prepared and purified from stan-dard strain PG3 to develop an indirect ELISA for detection of serum antibodies against M.mycoides subsp.capri.Results of chessboard titration test showed that the optimal concentration of coating antigen was 1.09 μg/mL,the optimal dilution of serum was 1∶64,the optimal dilution of HRP-IgG was 1∶40 000,and the optimal combination of antigen with antibody was for one hour.The criteria for evaluation was as follows:the serum was positive if P/N ≥3,the serum was negative if P/N≤2.5,and the serum was suspicious if P/ N is between 2.5 and 3.Results of reproducibility and stability tests confirmed that the developed assay had good stability and reproducibility.Compared with indirect haemagglutination(IHA),the indirect ELISA was more sensitive.更多还原