Veliceasa D, Tauscher G, Surányi G, Kós P B, Likó I, Santore U, Proll E, Ehrig F, Uray K, Hudecz F, Kühne T, Lukács N
Biological Research Center, Institute of Plant Biology, Szeged, Hungary.
Arch Virol. 2005 Dec;150(12):2501-12. doi: 10.1007/s00705-005-0589-7. Epub 2005 Jul 14.
Barley mild mosaic virus (BaMMV), a member of the family Potyviridae, genus Bymovirus, is involved in the economically important yellow mosaic disease of winter barley in East Asia and Europe. We investigated serological properties of bacterially expressed BaMMV coat protein (CP) of a German isolate. Ten mouse monoclonal antibodies were produced using purified E. coli expressed BaMMV-CP as immunogen. The reactivity of MAbs with different strains of BaMMV was analysed by several immunological methods that are frequently used in diagnostic virology: enzyme-linked immunosorbent assay (ELISA), dot-blot, Western-blotting (WB), direct tissue blotting immunoassay (DTBIA) and immunoelectron microscopy (IEM). The amino acids involved in the formation of epitopes recognised by several MAbs were mapped by using synthetic pin-bound peptides and the localisation of epitopes in assembled virus particles was determined by electron microscope studies. MAbs V29 and M1 decorated the whole virion indicating that their epitopes 6PDPI9 and 96ITDDEK101, respectively, are exposed on the surface. The MAbs V6 and V14 both interacted with 44LPEPKM49, which seems to be accessible at only one end of the virus particle. The MAbs V6, V14, V29 and M1 detected epitopes common to a wide range of BaMMV isolates and can therefore be used effectively in routine diagnostic tests for BaMMV from barley leaves. We suggest that MAbs M1, V6, V14 and V29 are most suitable for use in TAS-ELISA, V6, V14 and V29 for Western blotting and V29 and M1 for electron microscope serology.
大麦轻花叶病毒（BaMMV）是马铃薯Y病毒科大麦病毒属的成员，与东亚和欧洲冬大麦具有重要经济意义的黄花叶病有关。我们研究了德国分离株的细菌表达大麦轻花叶病毒外壳蛋白（CP）的血清学特性。以纯化的大肠杆菌表达的大麦轻花叶病毒CP作为免疫原，制备了10种小鼠单克隆抗体。通过诊断病毒学中常用的几种免疫学方法分析单克隆抗体与不同大麦轻花叶病毒株的反应性：酶联免疫吸附测定（ELISA）、斑点印迹、蛋白质免疫印迹（WB）、直接组织印迹免疫测定（DTBIA）和免疫电子显微镜（IEM）。使用合成的针结合肽对几种单克隆抗体识别的表位形成中涉及的氨基酸进行定位，并通过电子显微镜研究确定表位在组装病毒颗粒中的定位。单克隆抗体V29和M1标记了整个病毒粒子，表明它们的表位分别为6PDPI9和96ITDDEK101，暴露在表面。单克隆抗体V6和V14均与44LPEPKM49相互作用，该表位似乎仅在病毒粒子的一端可及。单克隆抗体V6、V14、V29和M1检测到了广泛的大麦轻花叶病毒分离株共有的表位，因此可有效地用于大麦叶片大麦轻花叶病毒的常规诊断检测。我们建议单克隆抗体M1、V6、V14和V29最适合用于TAS-ELISA，V6、V14和V29用于蛋白质免疫印迹，V29和M1用于电子显微镜血清学。
