de Avila A C, Huguenot C, Resende R de O, Kitajima E W, Goldbach R W, Peters D
Department of Virology, Agricultural University, Wageningen, The Netherlands.
J Gen Virol. 1990 Dec;71 ( Pt 12):2801-7. doi: 10.1099/0022-1317-71-12-2801.
Twenty tomato spotted wilt virus (TSWV) isolates were serologically compared in ELISA employing five different procedures using a rabbit polyclonal antiserum against nucleocapsid proteins (NuAbR) and mouse monoclonal antibodies (MAbs), two directed to nucleocapsid proteins (N1 and N2) and four directed to glycoproteins G1 to G4. All the antisera were raised against TSWV-CNPH1. The 20 isolates were differentiated into two distinct serogroups. Serogroup I consisting of 16 isolates strongly reacted with NuAbR. The other four isolates were poorly recognized by NuAbR and were placed in another serogroup, designated II. The panel of MAbs differentiated the TSWV isolates into three serotypes. The 16 isolates forming serogroup I reacted strongly with the MAbs generated and were identified as serotype I isolates. The four isolates which made up serogroup II were split into serotypes II and III. The serotype II isolates did not respond or responded poorly with MAbs N1, N2 and G3. The two other isolates placed in serotype III were recognized by N1 but not by N2 and G3. Two isolates became defective after several mechanical passages and failed to respond or responded very poorly with MAbs directed to glycoproteins. Our results show that ELISA employing polyclonal and monoclonal antisera is a useful tool to differentiate TSWV isolates and to detect defective forms. The results also strongly suggest that TSWV nucleocapsid proteins are less conserved than the glycoproteins.
使用针对核衣壳蛋白的兔多克隆抗血清（NuAbR）和小鼠单克隆抗体（MAb），通过五种不同程序在酶联免疫吸附测定（ELISA）中对20个番茄斑萎病毒（TSWV）分离株进行了血清学比较。其中两种单克隆抗体针对核衣壳蛋白（N1和N2），另外四种针对糖蛋白G1至G4。所有抗血清均针对TSWV-CNPH1制备。这20个分离株被分为两个不同的血清群。血清群I由16个分离株组成，它们与NuAbR强烈反应。另外四个分离株与NuAbR反应较弱，被归入另一个血清群，命名为血清群II。单克隆抗体组将TSWV分离株分为三种血清型。形成血清群I的16个分离株与所产生的单克隆抗体强烈反应，被鉴定为血清型I分离株。组成血清群II的四个分离株又分为血清型II和III。血清型II分离株对单克隆抗体N1、N2和G3无反应或反应较弱。归入血清型III的另外两个分离株可被N1识别，但不能被N2和G3识别。两个分离株在经过几次机械传代后出现缺陷，对针对糖蛋白的单克隆抗体无反应或反应非常弱。我们的结果表明，使用多克隆和单克隆抗血清的ELISA是区分TSWV分离株和检测缺陷型的有用工具。结果还强烈表明，TSWV核衣壳蛋白的保守性低于糖蛋白。
