Ludemann L R, Katz J B
United States Department of Agriculture, Animal and Plant Health Inspection Service, Ames, IA 50010.
Biologicals. 1994 Mar;22(1):21-7. doi: 10.1006/biol.1994.1004.
An enzyme-linked immunosorbent assay (ELISA) procedure was developed to assay the cytopathic and noncytopathic bovine viral diarrhea (BVD) virus strains used in inactivated vaccines licensed by the United States Department of Agriculture. The assay uses a biotin-labeled, staphylococcal protein A purified polyclonal BVD antibody (Bab) from a calf hyperimmuned against NADL, Singer, C24v, New York-1 (NY-1) strains and a field isolate. The Bab recognized the following reference strains of BVD virus: NADL; NY-1; C24v; Singer; and a field isolate. Monoclonal antibodies (Mab) directed against gp48 and gp53 of the Singer strain of BVD could detect only the Singer and the NY-1 strains. None of the Mab tested could differentiate between cytopathic and noncytopathic BVD virus strains. In vaccines containing multiple viral and bacterial components, the Bab was specific for the BVD fraction. Two vaccines not recognized by the Bab differed from the others in the type of adjuvant. The formation of antigen-adjuvant complexes during vaccine production may inhibit the ability of Bab to detect BVD antigens in an ELISA format. This ELISA procedure enables the detection of BVD antigens and demonstrates the potential for in vitro testing of inactivated BVD vaccines in place of the currently required host animal testing.
已开发出一种酶联免疫吸附测定（ELISA）方法，用于检测美国农业部许可的灭活疫苗中使用的致细胞病变和非致细胞病变的牛病毒性腹泻（BVD）病毒株。该测定使用一种生物素标记的、经葡萄球菌蛋白A纯化的多克隆BVD抗体（Bab），该抗体来自对NADL、Singer、C24v、纽约-1（NY-1）毒株和一株田间分离株进行超免疫的小牛。该Bab可识别以下BVD病毒参考毒株：NADL；NY-1；C24v；Singer；以及一株田间分离株。针对BVD Singer毒株的gp48和gp53的单克隆抗体（Mab）只能检测到Singer和NY-1毒株。所测试的Mab均无法区分致细胞病变和非致细胞病变的BVD病毒株。在含有多种病毒和细菌成分的疫苗中，Bab对BVD组分具有特异性。两种未被Bab识别的疫苗在佐剂类型上与其他疫苗不同。疫苗生产过程中抗原-佐剂复合物的形成可能会抑制Bab以ELISA形式检测BVD抗原的能力。这种ELISA方法能够检测BVD抗原，并证明了在体外检测灭活BVD疫苗以替代当前所需的宿主动物检测的潜力。
