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脑心肌炎病毒免疫测定法在细胞培养中应用酶标记病毒特异性单克隆抗体快速检测病毒、中和抗体及干扰素。

Application of immunoassay of encephalomyocarditis virus in cell culture with enzyme-labeled virus-specific monoclonal antibodies for rapid detection of virus, neutralizing antibodies, and interferon.

作者信息

Vlaspolder F, Harmsen T, van Veenendaal D, Kraaijeveld C A, Snippe H

机构信息

Department of Virology, University of Utrecht, The Netherlands.

出版信息

J Clin Microbiol. 1988 Dec;26(12):2593-7. doi: 10.1128/jcm.26.12.2593-2597.1988.

Abstract

Encephalomyocarditis virus (EMCV)-specific monoclonal antibody UM 21.1 labeled with horseradish peroxidase was used to detect EMCV in L-cell monolayers. This direct enzyme immunoassay of EMCV, performed in wells of 96-well plates, could be applied for various purposes, such as early detection of virus multiplication, determination of 50% tissue culture infective doses, and rapid titration of interferon and EMCV-neutralizing antibodies. Multiplication of EMCV is indicated by a rapid increase of the absorbance values measured against EMCV-infected L cells starting as early as 4.5 h after virus inoculation. The early rise of absorbance (i.e., virus multiplication) is inhibited by interferon, allowing its rapid titration. Preincubation of the virus inoculum with neutralizing antibodies also yielded decreased absorbance values. With the latter enzyme immunoassay for neutralizing antibodies, performed after an infection period of 8 h, antibody titers measured were comparable to those obtained with a conventional plaque reduction test. We assume that similar assays could be developed for other picornaviruses (e.g., polioviruses).

摘要

用辣根过氧化物酶标记的脑心肌炎病毒(EMCV)特异性单克隆抗体UM 21.1检测L细胞单层中的EMCV。这种在96孔板孔中进行的EMCV直接酶免疫测定法可用于多种目的,如早期检测病毒增殖、测定50%组织培养感染剂量以及快速滴定干扰素和EMCV中和抗体。早在病毒接种后4.5小时,针对感染EMCV的L细胞测量的吸光度值迅速增加,表明EMCV发生了增殖。吸光度的早期升高(即病毒增殖)受到干扰素的抑制,从而可以对干扰素进行快速滴定。病毒接种物与中和抗体预孵育也会使吸光度值降低。在8小时感染期后进行的后一种中和抗体酶免疫测定中,测得的抗体效价与用传统蚀斑减少试验获得的效价相当。我们认为可以针对其他小RNA病毒(如脊髓灰质炎病毒)开发类似的测定方法。

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