Li L, Giambrone J J, Panangala V S, Hoerr F J
Department of Poultry Science, Auburn University, Alabama 36849, USA.
Avian Dis. 1996 Apr-Jun;40(2):349-57.
Monoclonal antibodies (MAbs) were produced against the avian reovirus strain S1133. MAbs were isotyped and used to develop diagnostic tests. Splenocytes from immunized mice were screened by enzyme-linked immunosorbent assay (ELISA). Two hybridomas secreted MAbs against avian reovirus S1133. One MAb secreted IgG1 and the other secreted IgG2a. All MAb light chains were kappa Specificity of MAbs was tested against four avian reovirus strains: S1133, 1733, CO8, and 2408. Strains S1133, 1733, and 2408 viruses were in the same subtype; the CO8 virus belonged to a different subtype. The MAbs reacted with all reovirus strains by ELISA, dot blot, immunofluorescence assay, and immunoblotting. No MAb had neutralizing activity against the tested reoviruses. Immunoblot analysis showed the one MAb bound to protein sigma A with molecular weight of 39,000 Daltons for all reovirus strains. Another MAb bound to the protein sigma C with an approximate molecular mass of 32,000 Daltons. An indirect immunoperoxidase (IP) procedure was developed using a MAb to detect reovirus in formalin-fixed, paraffin-embedded tissues from infected chickens and chicken embryo fibroblast cell cultures. The IP test was simple, fast, and economical and enabled simultaneous evaluation of viral antigen-producing cells with tissue pathologic changes confirming that the reovirus caused the lesions.
制备了针对禽呼肠孤病毒S1133株的单克隆抗体（MAb）。对单克隆抗体进行了亚型鉴定并用于开发诊断测试。通过酶联免疫吸附测定（ELISA）筛选免疫小鼠的脾细胞。有两种杂交瘤分泌针对禽呼肠孤病毒S1133的单克隆抗体。一种单克隆抗体分泌IgG1，另一种分泌IgG2a。所有单克隆抗体的轻链均为κ链。针对四种禽呼肠孤病毒株测试了单克隆抗体的特异性：S1133、1733、CO8和2408。S1133、1733和2408病毒株属于同一亚型；CO8病毒属于不同亚型。这些单克隆抗体通过ELISA、斑点印迹、免疫荧光测定和免疫印迹与所有呼肠孤病毒株发生反应。没有单克隆抗体对测试的呼肠孤病毒具有中和活性。免疫印迹分析表明，一种单克隆抗体与所有呼肠孤病毒株分子量为39000道尔顿的σA蛋白结合。另一种单克隆抗体与分子量约为32000道尔顿的σC蛋白结合。利用一种单克隆抗体开发了一种间接免疫过氧化物酶（IP）方法，用于检测来自感染鸡和鸡胚成纤维细胞培养物的福尔马林固定、石蜡包埋组织中的呼肠孤病毒。IP试验简单、快速且经济，能够同时评估产生病毒抗原的细胞与组织病理变化，证实呼肠孤病毒引起了病变。
