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2μg/mL,血清稀释倍数为200倍,酶标抗体稀释倍数为1∶5000,阳性临界值为0.222,敏感性可达到1∶6400,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。
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 2μg/mL,the serum dilution multiple was 200 times,the enzyme⁃labeled antibody dilution multiple was 1∶5000,the positive critical value was 0.222,and the sensitivity could reach 1∶6400.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.
作者
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
LI Peng;LEI Mengyao;FENG Lili;WANG Zhenwei;GUAN Chunxiao;ZHENG Hongshuang;WANG Junru;WANG Liping;LI Yanjin;WU Xinyuan;LIU Xingyou;JIN Qianyue(School of Biological Engineering,Xinxiang University,Xinxiang 453003,China;College of Animal Science and Technology,Henan University of Science and Technology,Xinxiang 453003,China;Hebi Polytechnic,Hebi 458030,China;College of Veterinary Medicine,Henan Agricultural University,Zhengzhou 450002,China;Institute for Animal Health,Henan Academy of Agricultural Sciences/Key Laboratory of Animal Immunology,Zhengzhou 450002,China)
出处
《河南农业科学》
北大核心
2024年第7期133-141,共9页
Journal of Henan Agricultural Sciences
基金
河南省农业科学院杰出青年科技基金项目(2024JQ01)。
