Establishment of Indirect ELISA Antibody Detection Method Based on Penton Protein of Fowl Adenovirus Serotype 4

doi:10.15933/j.cnki.1004-3268.2024.07.015

Journal of Henan Agricultural Sciences ›› 2024, Vol. 53 ›› Issue (7): 133-141.DOI: 10.15933/j.cnki.1004-3268.2024.07.015

• Animal Husbandry and Veterinary Medicine • Previous Articles Next Articles

Establishment of Indirect ELISA Antibody Detection Method Based on Penton Protein of Fowl Adenovirus Serotype 4

LI Peng¹，LEI Mengyao^1,2，FENG Lili³，WANG Zhenwei^1,2，GUAN Chunxiao^1,2，ZHENG Hongshuang^1,2，WANG Junru^1,4，WANG Liping¹，LI Yanjin¹，WU Xinyuan¹，LIU Xingyou¹，JIN Qianyue⁵

（1.School of Biological Engineering，Xinxiang University，Xinxiang 453003，China；2.College of Animal Science and Technology，Henan University of Science and Technology，Xinxiang 453003，China；3.Hebi Polytechnic，Hebi 458030，
China；4.College of Veterinary Medicine，Henan Agricultural University，Zhengzhou 450002，China；5.Institute for Animal Health，Henan Academy of Agricultural Sciences/Key Laboratory of Animal Immunology，Zhengzhou 450002，China）

Received:2024-03-04 Published:2024-07-15 Online:2024-08-01

基于Penton 蛋白禽腺病毒血清4 型间接ELISA抗体检测方法的建立

李鹏¹，雷梦瑶^1,2，冯丽丽³，王振伟^1,2，管春晓^1,2，郑洪双^1,2，王俊茹^1,4，王利平¹，李炎锦¹，吴欣媛¹，刘兴友¹，金前跃⁵

（1.新乡学院生物工程学院，河南新乡 453003；2.河南科技学院动物科技学院，河南新乡 453003；3. 鹤壁职业技术学院，河南鹤壁 458030；4.河南农业大学动物医学院，河南郑州 450002；5.河南省农业科学院动物疫病防控研究所/动物免疫学重点实验室，河南郑州 450002）

通讯作者: 刘兴友（1963-），男，重庆人，教授，主要从事动物分子病毒学方面的研究。E-mail：liuxingyou@souhu.com 金前跃（1982-），男，浙江建德人，副研究员，博士，主要从事动物疫病的监测与防控研究。E-mail：1956089861@ qq.com
作者简介:李鹏（1976-），男，河南新乡人，副教授，博士，主要从事生猪健康养殖及疫病控制等研究。E-mail：lpwlp@aliyun.com
基金资助:
河南省农业科学院杰出青年科技基金项目（2024JQ01）

Abstract

Abstract: Penton protein is the main structural protein that constitutes the capsid of fowl adenovirus serotype 4（FAdV‑4），which has high conservation and good immunogenicity. An indirect ELISA antibody detection method based on the purified Penton protein of FAdV‑4 was established.The results showed that the optimal coating concentration of Penton protein was 2 μg/mL，the serum dilution multiple was 200 times，the enzyme‑labeled antibody dilution multiple was 1∶5 000，the positive critical value was 0.222，and the sensitivity could reach 1∶6 400. There was no cross‑reaction with infectious bronchitis virus，infectious bursal disease virus，and infectious laryngotracheitis virus，and the specificity was good.The coefficient of variation of inter‑batch and intra‑batch repeatability was less than 10%.The positive detection rate in 100 clinical serum samples was 77%，and the coincidence rate with the commercial kit was 98%.In summary，the ELISA antibody detection method established in this study can be used for the clinical detection of antibodies against FAdV‑4.

Key words: Fowl adenovirus serotype 4, Penton protein, Indirect ELISA, Antibody detection

摘要： Penton蛋白是构成禽腺病毒血清4型（Fowl adenovirus serotype 4，FAdV-4）衣壳的主要结构蛋白，具备高度保守性与良好的免疫原性。以纯化的Penton蛋白为包被抗原，建立一种基于禽腺病毒血清4型Penton蛋白的间接ELISA抗体检测方法。结果显示，Penton蛋白最佳包被质量浓度为2 μg/mL，血清稀释倍数为200倍，酶标抗体稀释倍数为1∶5 000，阳性临界值为0.222，敏感性可达到1∶6 400，与鸡传染性支气管炎病毒、鸡传染性法氏囊病病毒、鸡传染性喉气管炎病毒均无交叉反应，特异性良好，批间批内重复性变异系数均<10%，在100份临床血清中阳性检出率为77%，与商品化试剂盒检出符合率可达98%。综上，建立的ELISA抗体检测方法可以用于禽腺病毒4型临床抗体检测。

关键词: 禽腺病毒血清4型, Penton蛋白, 间接ELISA, 抗体检测

CLC Number:

S852.65

LI Peng, LEI Mengyao, FENG Lili, WANG Zhenwei, GUAN Chunxiao, ZHENG Hongshuang, WANG Junru, WANG Liping, LI Yanjin, WU Xinyuan, LIU Xingyou, JIN Qianyue. Establishment of Indirect ELISA Antibody Detection Method Based on Penton Protein of Fowl Adenovirus Serotype 4[J]. Journal of Henan Agricultural Sciences, 2024, 53(7): 133-141.

李鹏, 雷梦瑶, 冯丽丽, 王振伟, 管春晓, 郑洪双, 王俊茹, 王利平, 李炎锦, 吴欣媛, 刘兴友, 金前跃. 基于Penton 蛋白禽腺病毒血清4 型间接ELISA抗体检测方法的建立[J]. 河南农业科学, 2024, 53(7): 133-141.

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Establishment of Indirect ELISA Antibody Detection Method Based on Penton Protein of Fowl Adenovirus Serotype 4

基于Penton 蛋白禽腺病毒血清4 型间接ELISA抗体检测方法的建立

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