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一种用于检测抗登革病毒E蛋白和NS1蛋白单克隆抗体的免疫酶测定法

[An immunoenzyme assay for the detection of monoclonal antibodies against the E and NS1 proteins of the dengue virus].

作者信息

Vázquez S, Pupo M, Pelegrino J L, Morier L, Castillo A, Otero A, Guzmán M G

机构信息

Instituto de Medicina Tropical Pedro Kourí.

出版信息

Rev Cubana Med Trop. 1995;47(2):122-6.

PMID:9805082
Abstract

The ELISA was standardized to detect monoclonal antibodies of dengue virus proteins E and NS1. One indirect ELISA was applied, using C6-36 cells inoculated with the A-15 strain, isolated during the dengue 2 epidemic in 1981 as an antigen source. These cells were fixed in ELISA plates at a 200,000 cell/well concentration. A cell control in similar conditions was used. Specific monoclonal antibodies to both proteins were used to standardize the system. Studies at different incubation periods, to determine the highest expression moment of these proteins in the cell membrane, were carried out. The results show a full response at 72 hours postinoculation for both proteins; a 14.7 ng/mL sensitivity was obtained for the detection of NS1, and of 1.43 ng/mL for E protein. This system allows the monoclonal antibodies primary screening to dengue 2 virus E and NS1 proteins.

摘要

酶联免疫吸附测定(ELISA)经过标准化处理,用于检测登革病毒E蛋白和NS1蛋白的单克隆抗体。采用一种间接ELISA法,使用接种了1981年登革热2型流行期间分离的A - 15株病毒的C6 - 36细胞作为抗原来源。将这些细胞以每孔200,000个细胞的浓度固定在ELISA板中。使用处于类似条件下的细胞对照。使用针对这两种蛋白的特异性单克隆抗体对系统进行标准化。进行了不同孵育期的研究,以确定这些蛋白在细胞膜上的最高表达时刻。结果显示,接种后72小时两种蛋白均呈现完全反应;检测NS1的灵敏度为14.7 ng/mL,E蛋白的灵敏度为1.43 ng/mL。该系统可用于对登革热2型病毒E蛋白和NS1蛋白的单克隆抗体进行初步筛选。

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