Tomato yellow leaf curl geminivirus: Serological evaluation of an improved purification method
Tomato yellow leaf curl geminivirus: Serological evaluation of an improved purification method
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DOI：
10.1111/j.1365-2338.1995.tb01467.x
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发表时间：
1995-01-01
期刊：
影响因子：
--
通讯作者：
Luisoni, E.
中科院分区：
文献类型：
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作者：
Al-Bitar, L.;Luisoni, E.
番茄黄曲叶双生病毒是番茄生产的重要限制因素，目前正在扩展到新的领域。由于病毒纯化的困难和病毒颗粒的低抗原性，迄今为止只能生产低滴度的抗血清。为了获得更好的抗血清，必须改进纯化方法，这反过来又需要更好的方法来测量使用替代纯化程序获得的病毒的产量。为此，我们研究了四种类型的 ELISA 测试。其中，生物素-亲和素ELISA在所有条件下均获得了良好的结果；间接 ELISA 对浓缩病毒制剂给出了更好的结果，但对粗提物不起作用。两种 DAS-ELISA 类型在缀合程序上有所不同，给出的结果不太可靠。根据这些测试，提出的最佳纯化方法包括将匀浆与 Driselase（浸渍酶）在 37 摄氏度下孵育 24 小时，与之前使用的最佳方法相比，产量增加了 4-8 倍。本塞姆氏烟草被证实是最好的来源植物。
Tomato yellow leaf curl geminivirus is an important limiting factor for tomato production and is now expanding into new areas. Difficulties in virus purification and low antigenicity of virus particles have so far allowed the production only of low-titre antisera. To obtain better antisera, the purification method has to be improved, and this in turn requires better methods for measuring the yields of virus obtained using alternative purification procedures. For this purpose, we have investigated four types of ELISA test. Among these, biotin-avidin ELISA gave good results under all conditions; indirect ELISA gave better results with concentrated virus preparations but did not work with crude extracts. Two DAS-ELISA types, differing in conjugation procedures, gave less reliable results. On the bases of these tests, the best purification method proposed included a 24-h incubation of the homogenate at 37 degree C with Driselase (macerating enzymes), and gave a 4-8 fold increase in yield, compared with the best method previously used. Nicotiana benthamiana was confirmed to be the best source plant.