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用烟草赤星病菌的悬浮液免疫家兔，得到效价为１：６４Ｏ的抗血清，按硫酸该沉淀法和ＤＥＡＥ纤维素离子交换柱层析法提取抗体球蛋白ＩｇＧ，用ＥＬＩＳＡ间接法进行试验，结果表明：辣根过氧化物酶标记羊抗兔结合物的最适稀释度为１：１００，最适抗体球蛋白ＩｇＧ的浓度为４～６μｇ／ｍＬ。该抗体球蛋白ＩｇＧ对来自吉林省内的５个市县的８个烟草赤星病菌株全部呈阳性反应．而对Ａｌｔｅｒｎａｒｉａｃｕｕｃｕｍｅｒｉｎａ和Ａｌｔｅｒｎａｒｉａｐｏｒｒｉ则呈阴性反应，对种子分离得到的Ｆｕｓａｒｉｕｍｓｐ．和Ｐｈｙｌｌｏｓｔｉｃｔａｓｐ．等４种真菌及烟草野火病菌和Ｘａｎｔｈｏｍｏｎａｓｓｐ．均呈阴性反应，表明该抗体球蛋白ＩｇＧ的特异性很强。对７种不同浓度孢子悬液接种花器所获种子进行检测．均呈阳性反应，对来自３省６个品种１３份种子进行检测，８价呈阳性反应．５份呈阴性反应，与种子分高培养结果一致。用ＥＬＩＳＡ法检测烟草种子是否带有赤星病菌是一种快速而准确的方法。
Rabbits were immunized with a suspension of tobacco brown spot bacteria to obtain an antiserum with a titer of 1: 64O. Antibody globulin IgG was extracted by sulfuric acid precipitation and DEAE cellulose ion exchange column chromatography, The results showed that the optimum dilution of horseradish peroxidase labeled goat anti-rabbit conjugate was 1: 100, and the concentration of optimal antibody globulin IgG was 4 ~ 6μg / mL. The antibody globulin IgG showed positive reaction to all eight strains of tobacco brown spot disease from five cities and counties in Jilin province. While negative to Alternariacuucumerina and Alternariaporri, Fusariumsp isolated from seeds. And Phyllostictasp. 4 kinds of fungi and tobacco wildfire bacteria and Xanthomonasp. All showed negative reaction, indicating that the antibody globulin IgG is very specific. Seeds from 7 inoculated flower spores of 7 different concentrations of spore suspension were tested. All showed positive reaction. Thirteen seeds of 6 varieties from 3 provinces were detected, and the positive reaction was 8-fold. 5 were negative reaction, consistent with the results of high seed culture. It is a fast and accurate way to determine whether tobacco seed is infected with brown spot bacteria by ELISA.