中国兽医科学2025，Vol.55Issue(10)：1334-1340,7.DOI:10.16656/j.issn.1673-4696.2025.0188
牛冠状病毒家兔源多克隆抗体的制备与应用
Preparation and application of rabbit polyclonal antibodies against bovine coronavirus
摘要
Abstract
The newly emerging coronaviruses present extremely serious hazards to humans.Bovine coronavirus(BCoV)has the potential for zoonosis,and effective method is absent for the isolation,identification,and pathogen detection of BCoV.In this study,rabbit polyclonal antibodies that specifically react with BCoV were prepared,laying a technical foundation for the detection and identification of BCoV.BCoV was inactivated and then immunized rabbits to prepare polyclonal antibod-ies against the virus.After three rounds of immunization,the neutralizing titer was detected,and then the antibodies were purified.The purified polyclonal antibodies were used to indirect im-munofluorescence(IFA),Western-blot,and sandwich ELISA assays.The results showed that the rabbit polyclonal antibodies reacted with BCoV,and the neutralizing titer could reach to 1∶28,and when reacting with caprine coronavirus(cpCoV),it could reach to 1∶29.And the polyclonal antibodies were able to specifically react with different strains of BCoV and cpCoV in IFA.Western-blot analysis for BCoV and cpCoV revealed that a specific band appeared at approximately 50 kDa.In the sandwich ELISA,the rabbit polyclonal antibodies were used as the coating antibodies,which only specifical-ly reacted with BCoV and cpCoV,and cross-reaction was not observed with other viruses.The poly-clonal antibodies specifically bind to different strains of BCoV and cpCoV,and could be used in IFA and Western-blot analysis for isolation and identification of BCoV and cpCoV.The prepared polyclonal antibodies have the potential to used as coating antibodies in the double-antibody sandwich ELISA detection method for the two CoVs.This study lays a foundation for the pathogen detection and epi-demiological investigation of BCoV and cpCoV.关键词
牛冠状病毒/多克隆抗体/双抗体夹心ELISA/鉴定Key words
bovine coronavirus/polyclonal antibody/sandwich ELISA/identification分类
畜牧业引用本文复制引用
刘筱筱,余秋蓉,李霞,毛立,许信刚,李彬..牛冠状病毒家兔源多克隆抗体的制备与应用[J].中国兽医科学,2025,55(10):1334-1340,7.基金项目
国家重点研发计划项目(2023YFD1801302),江苏省重点研发计划项目(BE2022330) （2023YFD1801302）
兽医生物技术国家重点实验室开放基金项目(SKLVBF202106) （SKLVBF202106）
