POLYCLONAL ANTIBODIES FOR THE IMMUNODIAGNOSTIC OF BOVINE VIRUSES

Abstract The objective of the study was to produce polyclonal antibodies to bovine viruses and to test the reactivity in immunoassays such as immunofluorescence, immunoperoxidase and slot blot. Bovine viral diarrhea virus (BVDV), bovine respiratory syncytial virus (BRSV), bovine herpesvirus type 1, 2 and 5 (BoHV-1, BoHV-2 and BoHV-5) blue tongue virus (BTV) and vaccinia virus (VACV) were amplified in cell culture and used to immunize rabbits. Animals were immunized at regular intervals and the hyper immune serum produced was used as primary antibody in the immunoassays. The working dilution for each antiserum was determined by limiting serial dilution and varied from 1: 800 to 1: 51,200. The highest dilutions were observed for immunoperoxidase, when compared with immunofluorescence and slot blot. Dilutions lower than 1:800 were associated with the presence of nonspecific reactions. Anti-BoHV-1, -BoHV-5, -BVDV and -BRSV antisera reacted against heterologous isolates in immunofluorescence and immunoperoxidase assays. It is concluded that the produced polyclonal antibodies have high concentrations of virus-specific antibodies and are an alternative for use in immunoassays.

摘要 本研究旨在制备针对牛源病毒的多克隆抗体，并在免疫荧光、免疫过氧化物酶及斑点印迹等免疫检测方法中验证其反应性。牛病毒性腹泻病毒（BVDV）、牛呼吸道合胞体病毒（BRSV）、牛疱疹病毒1型、2型和5型（BoHV-1、BoHV-2和BoHV-5）、蓝舌病病毒（BTV）以及痘苗病毒（VACV）均在细胞培养中扩增，随后用于免疫家兔。实验动物按固定间隔接受免疫，所获得的高免血清被用作免疫检测的一抗。通过系列梯度稀释法确定各抗血清的工作稀释倍数，其范围为1:800至1:51200。相较于免疫荧光与斑点印迹检测，免疫过氧化物酶检测可采用的抗血清最高稀释倍数更高。当稀释倍数低于1:800时，易出现非特异性反应。抗BoHV-1、抗BoHV-5、抗BVDV及抗BRSV抗血清可在免疫荧光和免疫过氧化物酶检测中与异源毒株发生反应。综上，本研究制备的多克隆抗体含有高浓度的病毒特异性抗体，可作为免疫检测的备选试剂。