Peterson J D, Kim J Y, Melvold R W, Miller S D, Waltenbaugh C
Department of Microbiology-Immunology, Northwestern University Medical School, Chicago, IL 60611.
J Immunol Methods. 1989 Apr 21;119(1):83-94. doi: 10.1016/0022-1759(89)90384-0.
This paper examines the parameters necessary for the efficient measurement of anti-Theiler's murine encephalomyelitis virus (TMEV) antibodies in an affinity-dependent manner using a variation of a solid-phase particle concentration fluorescence immunoassay (PCFIA). By allowing antibody to react with fluorochrome-labelled virus in fluid phase and subsequently capturing the resulting virus-antibody complexes with anti-immunoglobulin coated polystyrene particles (fluid-phase PCFIA), the present assay allows for both greater sensitivity, specificity and preservation of conformational viral epitopes than do solid-phase immunoassays. Fluid-phase PCFIA proved to be a more rapid quantitative assay than ELISA and significantly diminished non-specific binding by both untreated and heat-inactivated normal mouse sera. This methodology also allowed us to perform competition assays and to determine the dissociation kinetics of anti-viral antibody preparations, investigations which cannot generally be performed as solid-phase immunoassays. Thus fluid-phase PCFIA is a rapid and efficient immunoassay with excellent reproducibility and great versatility.
本文研究了采用一种固相颗粒浓度荧光免疫分析（PCFIA）变体以亲和依赖方式有效检测抗泰勒氏鼠脑脊髓炎病毒（TMEV）抗体所需的参数。通过使抗体与液相中荧光染料标记的病毒反应，随后用抗免疫球蛋白包被的聚苯乙烯颗粒捕获形成的病毒 - 抗体复合物（液相PCFIA），本分析方法比固相免疫分析具有更高的灵敏度、特异性以及对病毒构象表位的保留能力。液相PCFIA被证明是一种比ELISA更快的定量分析方法，并且显著减少了未处理和热灭活的正常小鼠血清的非特异性结合。该方法还使我们能够进行竞争分析并确定抗病毒抗体制剂的解离动力学，而这些研究通常无法通过固相免疫分析进行。因此，液相PCFIA是一种快速高效的免疫分析方法，具有出色的重现性和广泛的适用性。
