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以提纯的黄瓜花叶病毒M株系(CMV-M)病毒粒子以及体外原核表达的外壳蛋白(CMV-CP)为抗原免疫健康家兔,制备了CMV-M多克隆抗体。采用间接酶联免疫吸附试验检测其效价,并结合Western-blot比较了两种多克隆抗体的灵敏度。结果表明:制备的两种CMV-M多克隆抗血清效价均达到1:25600;检测病汁液的灵敏度方面,以病毒粒子为抗原的多克隆抗血清达到1:3200,明显优于以原核表达蛋白作为抗原的抗血清1:800,蛋白印迹着色前者在样品量倍比稀释后免疫条带清晰,后者条带模糊甚至消失。
The CMV-M polyclonal antibody was prepared from purified CMV-M strain (CMV-M) virus particles and the CMV-CP antigen in vitro. The titer was tested by indirect enzyme-linked immunosorbent assay, and the sensitivity of the two polyclonal antibodies was compared with Western-blot. The results showed that the titer of the two CMV-M polyclonal antisera reached 1: 25600. For the detection of the sensitivity of diseased juice, the polyclonal antiserum with virus particles as antigen reached 1: 3200, which was significantly better than that of prokaryotic Protein as anti-serum antiserum 1: 800, Western blotting The former was doubled in the sample volume after the immunoblot was clear, the latter band fuzzy or even disappear.