河南农业科学 ›› 2024, Vol. 53 ›› Issue (1): 125-132.DOI: 10.15933/j.cnki.1004-3268.2024.01.014

• 畜牧·兽医 • 上一篇    下一篇

减蛋综合征病毒抗体胶体金免疫层析试纸条的研制及初步应用

宋亚鹏1,孙亚宁2,刘琳1,杨继飞2,李新生1,魏蔷2,张改平1,2   

  1. (1. 河南农业大学动物医学院,河南 郑州 450046;2.河南省农业科学院动物免疫学重点实验室,河南 郑州 450002)
  • 收稿日期:2023-07-12 出版日期:2024-01-15 发布日期:2024-02-27
  • 通讯作者: 张改平(1960-),男,河南内黄人,研究员,博士,主要从事重大疫病快速检测技术与疫苗研究。E-mail:zhanggaiping2003@163.com 魏蔷(1988-),女,河南驻马店人,助理研究员,博士,主要从事病原分子生物学研究。E-mail:chrysqiang@126.com
  • 作者简介:宋亚鹏(1984-),女,河南荥阳人,助理研究员,博士,主要从事动物分子病原学及免疫学研究。E-mail:songyapeng0371@163.com
  • 基金资助:
    国家自然科学基金青年基金项目(32002284,32102653);河南省农业科学院优秀青年科技基金项目(2022YQ22)

Development and Preliminary Application of Colloidal Gold Immunochromatography Test Strips for Egg Drop Syndrome Virus   Antibodies

SONG Yapeng1,SUN Yaning2,LIU Lin1,YANG Jifei2,LI Xinsheng1,WEI Qiang2,ZHANG Gaiping1   

  1. (1.College of Veterinary Medicine,Henan Agricultural University,Zhengzhou 450046,China;2.Key Laboratory of Animal Immunology,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China)
  • Received:2023-07-12 Published:2024-01-15 Online:2024-02-27

摘要: 为建立快速检测减蛋综合征病毒(EDSV)抗体的方法,基于EDSV的Fiber蛋白,制备快速检测EDSV抗体的免疫层析试纸条,为养禽业提供针对病原微生物的感染预警及免疫评估。利用胶体金标记EDSV Fiber蛋白抗原为探针,将羊抗鸡IgG和抗Fiber蛋白的抗体包被到硝酸纤维膜上,分别作为质控线和检测线,对家禽EDSV抗体水平进行监测。结果表明,制备的EDSV免疫层析试纸条具有高度特异性,与其他家禽病毒抗体无交叉反应,仅与EDSV抗体反应,在试纸条上产生可见的检测线;并且制备的试纸条具有高度稳定性,在室温条件下可至少保存6个月。应用制备的检测EDSV抗体的免疫层析试纸条对从河南、安徽、山东等地养鸡场采集的576份鸡血清样本检测,与血凝抑制试验(HI)结果进行平行比较,二者的Kappa值为0.878,具有良好一致性。综上,制备的检测ESDV抗体的免疫层析试纸条具有高度的特异性、敏感性、稳定性和易操作性,对EDSV抗体的临床检测具有实际应用价值。

关键词: 减蛋综合征病毒, Fiber蛋白, 免疫层析试纸, 血凝抑制试验

Abstract: In order to establish a method for rapid detection of egg drop syndrome virus(EDSV)antibody,an immunochromatographic test strip for rapid detection of EDSV antibody was prepared based on the EDSV Fiber protein,which could provide infection warning and immune evaluation for pathogenic microorganisms in poultry industry. Colloidal gold‑labeled EDSV Fiber protein was used as a probe,the goat anti‑chicken IgG and anti‑Fiber polyclonal antibody were blotted onto the nitrocellulose membrane as quality control lines and detection lines respectively to test EDSV antibody. The results showed that the EDSV immunochromatographic test strip was highly specific,had no cross‑reactivity with other viral antibodies,it only reacted with EDSV antibodies and produced a visible detection line on the test strip;and the test strips were highly stable and can be stored for at least 6 months at room temperature. The clinical application effect of the strips was tested by detecting EDSV antibodies in 576 serum samples collected from chickens farms in Henan,Anhui and Shandong,and compared with the hemagglutination inhibition test(HI)in parallel,and the Kappa value of the two was 0. 878,which was in good agreement. In summary,theimmunochromatographic test strips developed in this study are highly specific,sensitive,stable and easy to operate,and have practical application value for the clinical detection of EDSV antibodies.

Key words: word:Egg drop syndrome virus, Fiber protein, Immunochromatographic strip, Hemagglutination inhibition test(HI)

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