黄瓜花叶病毒（CMV）高效价抗血清的制备及其在FLISA中的应用  

THE PRODUCTION OF A HIGH-TITRE ANTISERUM OF CUCUMBER MOSAIC VIRUS AND ITS APPLICATION IN ELISA

作　　者：周正来[1] 陆仕华[1] 袁贤溶 汪树俊[1] 蒋震同 

机构地区：[1]上海市农业科学院植物保护研究所,上海农学院园林环境科学系

出　　处：《上海农学院学报》1995年第3期202-207,共6页

摘　　要：以提纯的ＣＭＶ作为抗原，经静脉注射、肌肉注射和足垫注射等对家兔进行免疫，获得高效价的抗血清（琼脂双扩散效价为１／２５６）；在间接ＥＬＩＳＡ试验过程中，用ＰＢＳ＋０．０５％ＴｒｉｔｏｎＸ－１００作为样本研磨液，用１：１６０００稀释度的ＣＭＶ抗血清或Ｚμｇ／ｍｌＣＭＶ抗血清的ＩｇＧ作为第一抗体，用１：１０００稀释度的冻干辣根过氧化物酶标记的羊抗兔的ＩｇＧ作为第二抗体，以待测样本光密度值与阴性对照光密度值之比大于或等于３作为判断待测样本为阳性反应的标准，结果表明：间接ＥＬＩＳＡ试验的灵敏度比生物检测的灵敏度要高３２～１６０倍。用间接ＥＬＩＳＡ检测矮牵牛的病毒样本３０份，ＣＭＶ占７０％。A high-titre antiserum of Cucumber Mosaic Virus which titre was determined as 1/256 by double diffusion test was obtained by immunizing rabbits with purified CMV as antigen through intravenous,intramuscular,and footpad injection. In indirect ELISA,PBS with 0.05% Triton X-100 was a solution inthe sample grind,the first antibody was rabbit antiserum to CMV (1:16000 dilution) or its purified IgG (2μg/ml),and the second antibody was the lyophilized HRP-labelled goat anti-rabbit-IgG(1:1000 dilution).A positive result was determined according to the absorbance ratio of tested sample to negative control,if it was equal to or more than 3.The results indicated that the detection sensitivity of indirect ELISA was 32～160 folds higher than that of the biological assays,and it was further confirmed by indirect ELISA that CMV was the prevailing virus in petunia in Shanghai.

关 键 词：黄瓜花叶病毒 家兔免疫 抗血清 ELISA 

分 类 号：S858.291.5[农业科学—临床兽医学] S436.421.1[农业科学—兽医学]