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一种通过细胞中线粒体脱氢酶功能测定(MTT 法)来精确量化干扰素活性的新型灵敏生物测定法。

A new sensitive bioassay for precise quantification of interferon activity as measured via the mitochondrial dehydrogenase function in cells (MTT-method).

作者信息

Berg K, Hansen M B, Nielsen S E

机构信息

Institute of Medical Microbiology, University of Copenhagen, Denmark.

出版信息

APMIS. 1990 Feb;98(2):156-62. doi: 10.1111/j.1699-0463.1990.tb01016.x.

Abstract

A biological method for precise quantification of interferons has been developed. The method is based upon the dehydrogenase system of the intact target cells, which will normally convert an artificial substrate, MTT, into formazan (blue), which, in turn, can be measured spectrophotometrically. This conversion is greatly reduced by cytocidal viruses in a dose-dependent manner. The protection of target cells by interferon against challenge virus is reflected in a diminished reduction in the production of formazan, thus giving a very precise method for quantification of interferon. The lowest level of detection is around 0.10 international units. The intra- and inter-assay variability appear to be below 10%. The assay, which makes no use of expensive ingredients, is performed in 96-well micro-trays and read in an inexpensive ELISA-scanner.

摘要

已开发出一种精确量化干扰素的生物学方法。该方法基于完整靶细胞的脱氢酶系统,该系统通常会将人工底物MTT转化为甲臜(蓝色),进而可通过分光光度法进行测量。这种转化会被杀细胞病毒以剂量依赖的方式大幅降低。干扰素对靶细胞免受攻击病毒的保护作用表现为甲臜生成量减少的程度降低,从而提供了一种非常精确的干扰素量化方法。最低检测水平约为0.10国际单位。实验内和实验间的变异性似乎低于10%。该检测方法不使用昂贵的成分,在96孔微量板中进行,并在廉价的酶联免疫吸附测定扫描仪中读取结果。

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