非洲猪瘟病毒p30抗体竞争化学发光法的建立  

Establishment of competitive chemiluminescence method for detection of African swine fever virus p30 antibody

作　　者：温升辉 邵军军[2] 高闪电[2] 彭德财 常惠芸[2] 丁嘉烽 刘伟 师铭咸 WEN Shenghui;SHAO Junjun;GAO Shandian;PENG Decai;CHANG Huiyun;DING Jiafeng;LIU Wei;SHI Mingxian(Laboratory of Clinical Veterinary Medicine,College of Animal Science and Technology,Guangxi University,Nanning 530004,China;National Key Laboratory of Animal Disease Prevention and Control/OIE/National Reference Laboratory for Foot-and-Mouth Disease,Gansu Provincial Research Center for Basic Discipline of Pathogenic Biology,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China)

机构地区：[1]广西大学动物科学技术学院临床兽医学实验室,广西南宁530004 [2]中国农业科学院兰州兽医研究所动物疫病防控全国重点实验室/甘肃省病原生物学基础学科研究中心OIE/国家口蹄疫参考实验室,甘肃兰州730046

出　　处：《中国兽医学报》2025年第1期1-7,共7页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(32273038);国家重点研发计划资助项目(2021YFD11801402)。

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∶2048 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。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 genotypeⅡstrains and the introduction of genotypeⅠattenuated 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(1:2048 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.

关 键 词：非洲猪瘟 p30抗体 抗体检测 化学发光法 

分 类 号：S852.65[农业科学—基础兽医学]