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2025年8月12日 星期二
农业生物技术学报  2022, Vol. 30 Issue (1): 185-193    DOI: 10.3969/j.issn.1674-7968.2022.01.017
  研究资源与技术改进 本期目录 | 过刊浏览 | 高级检索 |
非洲猪瘟病毒抗体快速检测试纸条的初步研制
江地科1,*, 邬旭龙2,*, 陈弟诗3, 王印1,**, 姚学萍1, 罗燕1, 杨泽晓1, 张鹏飞1
1四川农业大学 动物医学院/动物疫病与人类健康四川省重点实验室,成都 611130;
2成都农业科技职业学院,成都 611130;
3四川省动物疫病预防控制中心,成都 610041
Preliminary Development of a Test Strip for Rapid Detection of Antibodies Against African swine fever virus
JIANG Di-Ke1,*, WU Xu-Long2,*, CHEN Di-Shi3, WANG Yin1,**, YAO Xue-Ping1, LUO Yan1, YANG Ze-Xiao1, ZHANG Peng-Fei1
1 College of Veterinary Medicine/Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu 611130, China;
2 Chengdu Agricultural College, Chengdu 611130, China;
3 Sichuan Center for Animal Disease Control and Prevention, Chengdu 610041, China
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摘要 为建立一种能快速、高效、简便检测非洲猪瘟病毒(African swine fever virus, ASFV)抗体的方法,本研究采用原核表达系统表达ASFV衣壳蛋白(p72)作为捕捉抗原,以纯化的抗ASFV p72蛋白多克隆抗体和兔抗猪IgG分别包被于硝酸纤维素膜上,分别作为质控线(C线)和检测线(T线),优化反应条件,制备胶体金免疫层析试纸条(colloidal gold immunochromatographic test strip, ICTS),用于临床样品检测。结果表明,本研究成功构建pCold-TF-p72重组蛋白,经Western blot和琼脂扩散实验表明,制备的抗p72多克隆抗体具有良好的反应原性与免疫原性;制备的胶体金试纸条与针对猪伪狂犬病病毒(Pseudorabies virus, PRV)、猪圆环病毒2型(Porcine circovirus 2, PCV2)、猪繁殖与呼吸综合征病毒(Porcine reproductive and respiratory syndrome virus, PRRSV)、猪瘟病毒(Classical swine fever virus, CSFV)阳性血清无交叉反应,具有良好的特异性;临床样品阳性检出率为55.17% (48/87),商品ELISA试剂盒阳性检出率为59.77% (52/87),两者阳性符合率为92.31% (48/52)。本研究制备的胶体金试纸条为临床ASFV抗体检测及疫病预防具有一定的参考意义。
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江地科
邬旭龙
陈弟诗
王印
姚学萍
罗燕
杨泽晓
张鹏飞
关键词 非洲猪瘟病毒(ASFV)p72蛋白原核表达胶体金    
Abstract:In order to establish a rapid, efficient and simple method for detecting African swine fever virus (ASFV) antibodies, this study used a prokaryotic expression system to express ASFV capsid protein (p72) as a capture antigen, and anti-p72 polyclonal antibody and rabbit anti-pig IgG were coated on nitrocellulose membrane, as the quality control line (C line) and detection line (T line), respectively. The reaction conditions were optimized to prepare colloidal gold immunochromatographic test strip (ICTS), used for clinical sample detection. The results showed that the pCold-TF-p72 recombinant protein was successfully constructed in this study. Western blot and agar expansion experiments showed that the prepared anti-p72 polyclonal antibody had good reactogenicity and immunogenicity. The test strips could specifically detect anti-ASFV antibody and had no cross-reaction with positive sera against Pseudorabies virus, Porcine circovirus 2, Porcine reproductive and respiratory syndrome virus and Classical swine fever virus; In the detection of clinical samples, the positive detection rate of ICTS was 55.17% (48/87), the positive detection rate of a commercial ELISA kit was 59.77% (52/87), respectively. The agreement between ICTS and a commercial ELISA kit (IDVET) was 92.31% (48/52). The ICTS prepared in this study provides a reference for clinical ASFV antibody detection and disease prevention.
Key wordsAfrican swine fever virus (ASFV)    p72 protein    Prokaryotic expression    Colloidal gold
收稿日期: 2021-03-05     
ZTFLH:  S855.3  
基金资助:四川省科技计划(2021YJ0270); 2020年成都市科技项目技术创新研发项目(2019-YF05-00119-SN); 成都农业科技职业学院2020年院级科研基金(20ZR101)
通讯作者: **yaanwangyin@tom.com   
作者简介: * 同等贡献作者
引用本文:   
江地科, 邬旭龙, 陈弟诗, 王印, 姚学萍, 罗燕, 杨泽晓, 张鹏飞. 非洲猪瘟病毒抗体快速检测试纸条的初步研制[J]. 农业生物技术学报, 2022, 30(1): 185-193.
JIANG Di-Ke, WU Xu-Long, CHEN Di-Shi, WANG Yin, YAO Xue-Ping, LUO Yan, YANG Ze-Xiao, ZHANG Peng-Fei. Preliminary Development of a Test Strip for Rapid Detection of Antibodies Against African swine fever virus. 农业生物技术学报, 2022, 30(1): 185-193.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2022.01.017     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2022/V30/I1/185
 
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