The P54 protein of African swine fever virus（ASFV） was prepared with gene recombination technique and used to immune Balb/C mice, the spleen cells of immunized mice was fused with SP2/0, cloned by limiting dilution thus obtaining hybridoma cell lines secreting anti-ASFV antibody stably. An ELISA for the detection of ASFV was established using the monoclonal antibody. The results showed that the ELISA kit could be used for the detection of African swine fever antibody with high specificity and no cross-reaction. The development of the ELISA method had important practical significance for the laboratory diagnosis and epidemiological investigation of ASFV.%用基因重组技术制备的非洲猪瘟蛋白P54免疫BALB/C小鼠,将免疫鼠的脾细胞与SP2/0骨髓瘤细胞融合,采用有限稀释法进行克隆,获得能稳定分泌抗ASFV单克隆抗体的杂交瘤细胞株。使用该单克隆抗体,建立了检测血清中非洲猪瘟抗体的竞争法ELISA。实验结果表明：ELISA竞争法特异性高,无交叉反应,灵敏度高于间接免疫荧光法,可用于猪血清的非洲猪瘟抗体检测。该法的建立对非洲猪瘟实验诊断的标准以及流行病学调查具有重要的现实意义。
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