PRODUCTION OF MONOCLONAL-ANTIBODIES TO AFRICAN CASSAVA MOSAIC-VIRUS AND DIFFERENCES IN THEIR REACTIVITIES WITH OTHER WHITEFLY-TRANSMITTED GEMINIVIRUSES
PRODUCTION OF MONOCLONAL-ANTIBODIES TO AFRICAN CASSAVA MOSAIC-VIRUS AND DIFFERENCES IN THEIR REACTIVITIES WITH OTHER WHITEFLY-TRANSMITTED GEMINIVIRUSES
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DOI：
10.1099/0022-1317-67-12-2739
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发表时间：
1986-12-01
影响因子：
3.8
通讯作者：
HARRISON, BD
中科院分区：
文献类型：
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作者：
THOMAS, JE;MASSALSKI, PR;HARRISON, BD
用间接ELISA法筛选出一组抗非洲木薯花叶病毒（ACMV-JI）的鼠源单克隆抗体。在从腹水中纯化的10种单克隆抗体中，5种中有4种产生了更强的多克隆抗体反应。10个单克隆抗体中的6个在免疫扩散试验中产生沉淀反应，9个在免疫吸附电镜中捕获ACMV-JI颗粒。兔抗ACMV多克隆抗体在DAS-ELISA中不仅能与ACMV-JI和ACMV肯尼亚海岸株（ACMV-C）反应，而且能与其它5种已知或疑似以烟粉虱为载体的双生病毒反应，但不能与3种以叶蝉为载体或未知载体的双生病毒反应。在10株单抗中，有4株单抗在间接ELISA中不与ACMV-C反应，只有2株单抗与ACMV-C反应强烈。这支持了ACMV-C是ACMV的一个独特菌株而不仅仅是一个微小变异的其他证据。其他粉虱传播的双生病毒（菜豆金花叶病毒，大戟花叶病毒，绿豆黄花叶病毒和番茄金花叶病毒）和澳大利亚番茄曲叶病毒与2至5个单抗反应，但每种病毒有不同的反应模式。与此相反，茄顶端卷叶病毒，叶蝉传播的甜菜曲顶和玉米条纹病毒，不与任何单克隆抗体。竞争性结合试验的结果，当结合与不同的病毒的反应模式的个别单克隆抗体，表明单克隆抗体是特异性的9个不同的表位。两种单克隆抗体与所有五种白色传播的病毒反应，表明这些单克隆抗体检测到的表位可能是重要的传播B。烟粉虱单个单克隆抗体似乎适合检测和鉴定ACMV，用于区分ACMV-JI和ACMV-C，以及用于其他粉虱传播的双生病毒的定量测定。
A panel of murine monoclonal antibodies (MAbs), selected by indirect ELISA, was prepared to an isolate (ACMV-JI) of the type strain of African cassava mosaic virus. Of the ten MAbs purified from ascitic fluids, four out of the five tested gave stronger reactions polyclonal antibody. Six of the ten MAbs gave a precipitin reaction in immunodiffusion tests and nine trapped ACMV-JI particles in immunosorbent electron microscopy. Rabbit polyclonal antibody to ACMV reacted in DAS-ELISA not ony with ACMV-JI and the Kenya coast strain of ACMV (ACMV-C) but also with five other geminiviruses known or suspected to have the whitefly Bemisia tabaci as a vector, though not with thre geminiviruses that have leafhopper or unknown vectors. Of ten MAbs studied in detail, four did not react with ACMV-C in indirect ELISA and only two reacted strongly. This supports other evidence that ACMV-C is a distinctive strain of ACMV and nor merely a minor variant. The other whitefly-transmitted geminiviruses (bean golden mosaic, euphorbia mosaic, mung bean yellow mosaic and tomato golden mosaic viruses) and Australian tomato leaf curl virus reacted with two to five MAbs but each virus had a different pattern of reactivity. In contrast, solanum apical leaf curling virus, and the leafhopper-transmitted beet curly top and maize streak viruses, did not react with any of the MAbs. The results of competitive binding tests, when combined with the patterns of reaction of individual MAbs with different viruses, indicated that the MAbs were specific for nine distinct epitopes. Two MAbs reacted with all five whitely-transmitted viruses, suggesting that the epitopes detected by these MAbs may be important for transmission by B. tabaci. Individual MAbs seem suitable for detecting and identifying ACMV, for distinguishing between ACMV-JI and ACMV-C, and for quantitative assays of other whitefly-transmitted geminiviruses.