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南方菜豆花叶病毒(SBMV)、烟草花叶病毒(TMV)经 PEG 沉淀、差速离心、梯度离心纯化后,以每只鼠0.1、0.5、1 0、1.5、2mg 剂量,每周一次,共三次分别免疫小鼠,末次免疫后8天注射鼠腹水癌细胞(S_(180)),8～13天后鼠腹部膨大,抽取腹水,离心,取上清液,即为特异腹水抗体。实验证明:每只小鼠免疫0.1mg 抗原,获得抗体滴度1:200万(ELISA);免疫1.5mg,可获得最高滴度1:3200万的特异抗体。同时,还研制了大豆花叶病在毒(SMV)、马铃薯×病毒(PVX),马铃薯 Y 病毒(PVY),蚕豆萎蔫病毒(BBWV)、花生矮花病毒(PSV)、芜菁花叶病毒(T_uMV)腹水抗体,滴度为1:400万～3200万。一般一只小鼠可以获得20～40ml,最多66ml 腹水抗体,其抗体性质与同种抗原免疫兔、鼠制备的抗血清相同。这是用微量抗原制备多量特异抗体的好方法。用鼠腹水抗体与兔抗血清组合异种动物抗体双夹心酶联试验,可得到与经典的双抗体夹心法相当或略好的检测效果,又不需自制特异抗体酶标记物,适用于检测大量样品,为植物检疫提供一个好的检验手段。
Southern bean mosaic virus (SBMV) and tobacco mosaic virus (TMV) were isolated by PEG precipitation, differential centrifugation and gradient centrifugation. The mice were treated with 0.1, 0.5, The mice were respectively immunized three times, and the mice were injected with ascites carcinoma cells (S_ (180)) 8 days after the last immunization. After 8 ~ 13 days, the mice ’s abdomen was swollen and the ascites was aspirated, centrifuged and the supernatant was taken. Experiments show that: each mouse immunized 0.1mg antigen, antibody titer 1: 2 million (ELISA); immune 1.5mg, get the highest titer of 1: 32000000 specific antibodies. At the same time, the soybean mosaic virus (SMV), PVX, PVY, BBWV, PSV, T_uMV) ascites antibody titer of 1: 4 million ~ 32 million. Generally a mouse can get 20 ~ 40ml, up to 66ml ascites antibody, the antibody properties and alloantigens immunized rabbits, mouse antisera prepared the same. This is a good way to prepare large amounts of specific antibodies with trace amounts of antigens. The combination of murine ascites antibody and rabbit antiserum in combination with heterologous antibody double-sandwich enzyme-linked test can be obtained with the classic double antibody sandwich method, the detection effect is equivalent or slightly better, but also does not require self-made specific antibody enzyme markers, suitable for testing a large number of samples , To provide a good inspection method for plant quarantine.