Hajimorad M R, Francki R I
Department of Plant Pathology, Waite Agricultural Research Institute, University of Adelaide, Australia.
Arch Virol. 1991;117(3-4):219-35. doi: 10.1007/BF01310767.
The adsorption and retention properties of native (unfixed) and glutaraldehyde-fixed alfalfa mosaic virus (AMV) antigens to the polystyrene of ELISA plates were studied using [35S]-labelled virus preparations. It was shown that adsorption was a temperature-dependent, relatively slow process which varied between different AMV isolates. The amount of virus antigen adsorbed was dependent on the type and pH of the suspending buffer. Although native virus antigen adsorbed very efficiently at high pH when the particles had dissociated, significant amounts also adsorbed at pH 7.0, or lower. However, glutaraldehyde-fixed virus particles which retained their integrity even at pH as high as 9.6, adsorbed much more efficiently than native virus antigen above pH 9.0, but hardly at all around pH 7.0. The wide variation in adsorption of AMV antigen to microtitre plates under even slightly different conditions had significant influence on ELISA readings, which calls for extreme caution in interpreting serological results from indirect ELISA when antigen is used to coat the microtitre plates.
利用[35S]标记的病毒制剂，研究了天然（未固定）和戊二醛固定的苜蓿花叶病毒（AMV）抗原对酶联免疫吸附测定（ELISA）板聚苯乙烯的吸附和保留特性。结果表明，吸附是一个依赖温度的相对缓慢的过程，不同的AMV分离株之间存在差异。吸附的病毒抗原量取决于悬浮缓冲液的类型和pH值。尽管当颗粒解离时，天然病毒抗原在高pH值下吸附非常有效，但在pH 7.0或更低时也有大量吸附。然而，即使在高达pH 9.6的情况下仍保持完整性的戊二醛固定病毒颗粒，在pH 9.0以上比天然病毒抗原吸附效率高得多，但在pH 7.0左右几乎不吸附。即使在稍有不同的条件下，AMV抗原对微量滴定板的吸附差异很大，这对ELISA读数有显著影响，这就要求在使用抗原包被微量滴定板时，在解释间接ELISA的血清学结果时要格外谨慎。
