抗鹅细小病毒单克隆抗体的制备及其免疫荧光检测方法的建立

摘要

In order to develop an indirect immunofluorescent assay (IFA) for detection of Goose parvovirus (GPV), we generated three monoclonal antibodies (GPV-Mab-6C8, GPV-Mab-1B9 and GPV-Mab-2H8) using ultracentrifugation-purified virions for mouse immunization. All of three Mabs were specific to GPV and had virus neutralization activities. An IFA method was developed using these Mabs and used to detect viruses in the infected duck embryo fibroblast cultures and frozen tissue sections. Results showed that the established IFA method can be used in clinical diagnosis of GPV and titration of vaccines.%　　为建立免疫荧光检测鹅细小病毒（Goose parvovirus，GPV）的方法，本研究利用超速离心纯化的GPV作为免疫原，制备了3株抗GPV特异性单克隆抗体，依次命名为GPV-Mab-6C8、GPV-Mab-1B9、GPV-Mab-2H8。试验结果表明，3株抗GPV抗体特异性良好，并能够中和GPV对鹅胚成纤维细胞的感染能力。利用抗GPV单抗作为一抗，FITC标记的羊抗鼠IgG为二抗，建立了检测GPV的间接免疫荧光方法（indirect immunofluorescence assay, IFA），该方法不仅能够检测鹅胚成纤维细胞中的GPV，还能够检测发病鹅组织冰冻切片中的GPV。本研究制备的抗GPV特异性单克隆抗体和建立的检测GPV的免疫荧光方法，为今后GPV细胞活疫苗的检测评价和临床诊断提供了技术手段。