数字农科院2.0

Development and Application of Indirect ELISA for IBDV VP2 Antibodies Detection in Poultry

文献类型： 外文期刊

作者： Wenying Zhang;Yulong Wang;Guodong Wang;Hangbo Yu;Mengmeng Huang;Yulong Zhang;Runhang Liu;Suyan Wang;Hongyu Cui;Yanping Zhang;Yuntong Chen;Yulong Gao;Xiaole Qi

作者机构：

关键词： antibody detection;ELISA;IBDV;VLP;VP2

期刊名称： Viruses

ISSN： 1999-4915

年卷期： 2025 年 17 卷 7 期

页码：

收录情况： SCIE（2025版）

摘要： Infectious bursal disease virus (IBDV) is one of the most important immunosuppressive viruses in poultry, causing the global spread of infectious bursal disease (IBD). It poses a significant threat to the healthy development of the poultry industry. Vaccination is an effective approach for controlling IBDV infection. Therefore, reliable immune monitoring for IBDV is critical for maintaining poultry health. The enzyme-linked immunosorbent assay (ELISA) is a common technique used to detect specific antibodies in clinical serum testing and for the serological evaluation of IBDV vaccines. Among the currently available and under development IBDV vaccines, IBD VP2 subunit-based vaccines account for a considerable proportion. These vaccines stimulate the production of antibodies that are specific only to VP2. However, most IBDV antibody ELISA kits approved for use have applied the whole virus as the coating antigen, which does not adequately meet the diverse requirements for IBDV detection across different conditions. This study utilized a prokaryotic expression system to express the VP2 protein of the IBDV epidemic strain, assembling it into virus-like particles to be used as coating antigens. This approach enabled the establishment of an indirect ELISA method for detecting IBDV VP2 antibody (VP2-ELISA). The optimal coated antigen concentration was determined to be 2.5 μg/mL, with overnight coating at 4 °C; sealing with 5% skim milk at 37 °C for 4 h; serum dilution at 1:500 with incubation at 37 °C for 30 min; secondary antibody dilution at 1:4000 with incubation at 37 °C for 40 min; and then incubation with the substrate solution 3,3′,5,5′-tetramethylbenzidine at room temperature for 20 min. The criterion for interpreting the detection results was OD450nm ≥ 0.111 indicates IBDV antibody positivity, while OD450nm < 0.111 indicates negativity. The established VP2-ELISA can specifically detect IBDV-positive sera at the lowest serum dilution of 1:6400, with intra- and inter-batch coefficients of variation of <2%. This indicates that the VP2-ELISA exhibits good specificity, sensitivity, and stability. Detection experiments using 20 laboratory-immunized chicken serum samples and 273 clinical serum samples demonstrated that the results of VP2-ELISA were consistent with those of commercial ELISA kits coated with whole virus. In summary, the VP2-ELISA developed in this study offers advantages in immune response detection for IBD VP2 subunit-based vaccines and is appropriate for evaluating the efficacy of IBD vaccines and detecting clinical serum samples.

分类号：

  • 相关文献

[1]IBDV超强毒HZ株VP2基因的克隆和真核表达质粒的构建. 许信刚,李健强,王笑梅,童光志. 2000

[2]IBDV超强毒HZ株VP2基因的克隆和真核表达质粒的构建. 许信刚,李健强,王笑梅,童光志. 2000

[3]酪蛋白激酶CK1α通过与病毒蛋白VP2互作影响IBDV复制. 陈玉明,张礼洲,王念,卢珍,高立,王永强,高玉龙,高宏雷,刘长军,崔红玉,张艳萍,王笑梅,祁小乐. 2014

[4]The down-regulation of casein kinase 1 alpha as a host defense response against infectious bursal disease virus infection. Zhang, Lizhou,Li, Hui,Chen, Yuming,Gao, Xiang,Lu, Zhen,Gao, Li,Wang, Yongqiang,Gao, Yulong,Gao, Honglei,Liu, Changjun,Cui, Hongyu,Zhang, Yanping,Pan, Qing,Qi, Xiaole,Wang, Xiaomei,Wang, Xiaomei.

[5]OASL suppresses infectious bursal disease virus replication by targeting VP2 for degrading through the autophagy pathway. Wang, Suyan,Xu, Zhuangzhuang,Liu, Yongzhen,Yu, Mengmeng,Zhang, Tao,Liu, Peng,Qi, Xiaole,Chen, Yuntong,Meng, Lingzhai,Guo, Ru,Zhang, Li,Fan, Wenrui,Gao, Li,Duan, Yulu,Zhang, Yanping,Cui, Hongyu,Gao, Yulong. 2024

[6]Immunogenicity and protection against infectious bursal disease via a transgenic Eimeria acervulina expressing IBDV VP2-2C3d fusion protein. Yingying Sun,Yuehui Liu,Haodi Wang,Qingbin Guo,Jingxia Suo,Sixin Zhang,Xinming Tang,Guangwen Yin,Xun Suo,Xianyong Liu. 2025

[7]A recombinant Marek's disease vaccine candidate provides complete protection against infectious bursal disease virus and H9 subtype avian influenza virus in chickens. Fan, Wenrui,Zeng, Xianying,Chen, Yuntong,Yu, Qingqing,Zhang, Zibo,Tian, Guobin,Liu, Changjun,Bao, Hongmei,Qi, Xiaole,Wu, Longbo,Zhang, Yanping,Liu, Yongzhen,Wang, Suyan,Cui, Hongyu,Duan, Yulu,Chen, Hualan,Gao, Yulong. 2025

[8]Development of an Indirect ELISA for REV gp90 Antibody Detection Using the gp90 Protein Expressed in Suspended Cells. Erjing Ke,Mengmeng Huang,Guodong Wang,Jingzhe Han,Yulong Zhang,Runhang Liu,Hangbo Yu,Ziwen Wu,Dan Ling,Xianyun Liu,Tengfei Xu,Suyan Wang,Yuntong Chen,Yongzhen Liu,Yanping Zhang,Hongyu Cui,Yulu Duan,Liuan Li,Xiaoxue Yu,Yulong Gao,Xiaole Qi. 2026

[9]Development of an indirect ELISA using a novel linear epitope at the C-terminal region of the VP2 protein to specifically detect antibodies against Senecavirus A. Zhongyuan Ma,Jianliang Lv,Zhongwang Zhang,Li Pan. 2022

[10]猪细小病毒AV30 VP2基因在昆虫细胞中表达及其免疫原性鉴定. 李茜,代军飞,丁耀忠,ASHENAFI Kiros Wubshe,李国秀,马炳,张杰. 2020

[11]初步构建AIV、NDV、IBDV和IBV同步检测DNA芯片. 王秀荣,石锐,李冬梅,刘丽玲,孙凯,孔宪刚,包红梅,陈化兰. 2005

[12]IBDV反向遗传操作系统的建立和基因功能研究. 祁小乐,高玉龙,高宏雷,邓小芸,步志高,王晓燕,秦立廷,王笑梅. 2007

[13]抗传染性法氏囊病病毒VP5蛋白单克隆抗体的制备与初步应用. 张宁,高宏雷,高玉龙,李俊山,王晓艳,冉多良,王笑梅. 2007

[14]雏鸡混合感染IBDV与CAV的研究. 王笑梅,王秀荣,邱冬,王积海. 2000

[15]IBDV超强毒株Gx基因组大片段在大肠杆菌中的表达. 高宏雷,王笑梅,张厚双,付朝阳,王英,宋素泉. 2003

[16]高尔基体蛋白CSGalNAcT2利于IBDV衣壳蛋白成熟并促进病毒增殖. 张礼洲,高玉龙,任宪刚,陈玉明,王念,卢珍,高立,秦立廷,王永强,高宏雷,李凯,祁小乐,王笑梅. 2014

[17]鸡法氏囊B淋巴细胞全长cDNA表达文库的构建. 张礼洲,祁小乐,高玉龙,王笑梅. 2012

[18]IBDV强、弱毒VP2诱饵载体构建及在酵母双杂交系统中自激活作用的检测. 李铁强,高玉龙,高宏雷,邓小芸,王笑梅,王立群. 2008

[19]犬细小病毒VP2基因克隆及原核表达分析. 卫巧林,张韵,甄士伟,殷相平. 2016

[20]传染性法氏囊病病毒超强毒株致弱的分子生物学特征. 许信刚,王笑梅,李健强,童光志. 2000

作者其他论文 更多>>