【摘要】 非洲猪瘟(African swine fever, ASF)是由非洲猪瘟病毒(African swine fever virus, ASFV)感染引起猪的一种急性、热性、高致死性疾病。鉴于当前尚缺乏商业化的疫苗，并且近年来ASFV不断演变，中等毒力基因Ⅱ型毒株的涌现以及基因Ⅰ型弱毒株的传入，导致猪仅出现了持续性和慢性感染的状况。因此，对ASFV的特异性抗体检测变得势在必行。本研究利用p30单克隆抗体建立了一种检测ASFV p30抗体的竞争化学发光法——p30-cCLIA。通过检测背景清晰的阴阳性血清，确定该方法的Cut-off值为50%时，诊断敏感性(Dsn)和诊断特异性(Dsp)均为100%。在最佳反应条件下筛选出适用于p30单抗16-5E7E8-HRP的酶标稳定剂。此外，本研究建立的p30-cCLIA方法的灵敏度比商品化阻断ELISA试剂盒更高(1∶2 048 vs 1∶512),并具有良好的重复性。通过检测其他病毒感染猪阳性血清，结果表明与这些血清无交叉反应性。该方法的建立为ASF早期诊断提供了有力工具。更多还原

【Abstract】 African swine fever(ASF) is an acute,febrile,and highly fatal disease caused by African swine fever virus(ASFV) in pigs.Given the current lack of commercial vaccines and the continuous evolution of ASFV in recent years,the emergence of moderately virulent genotype Ⅱ strains and the introduction of genotype Ⅰ attenuated strains have led to persistent and chronic infections in pigs.Therefore,the detection of specific antibodies against ASFV has become imperative.In this study,we established a competitive chemiluminescence immunoassay(p30-cCLIA) for detecting ASFV p30 antibodies using p30 monoclonal antibodies.By detecting sera with clear negative and positive background,we determined that the Cut-off value of this method was 50%,with both diagnostic sensitivity(Dsn) and diagnostic specificity(Dsp) reaching 100%.Under optimal reaction conditions,we screened out an enzyme-labeled stabilizer suitable for p30 monoclonal antibody 16-5E7E8-HRP.Furthermore,the sensitivity of the established p30-cCLIA method was higher than that of the commercial blocking ELISA kit(1:2 048 vs 1:512) and exhibited good repeatability.Detection of sera positive for other porcine virus infections showed no cross-reactivity.The establishment of this method provides a powerful tool for early diagnosis of ASF.更多还原