Detection of hepatitis C virus specific core protein in serum of patients by a sensitive fluorescence enzyme immunoassay (FEIA)
Detection of hepatitis C virus specific core protein in serum of patients by a sensitive fluorescence enzyme immunoassay (FEIA)
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DOI：
10.1016/0022-1759(95)00261-8
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发表时间：
1996-03-28
影响因子：
2.2
通讯作者：
Kohara, M
中科院分区：
文献类型：
--
作者：
Kashiwakuma, T;Hasegawa, A;Kohara, M
使用抗重组丙型肝炎病毒（HCV）核心蛋白的单克隆抗体（mAb）开发了蛋白质捕获荧光酶免疫分析（FEIA），建立并表征了四种杂交瘤细胞系（5E3、5F11、515S、1080S）。这些单克隆抗体 (mAb) 各自具有 IgG1 和 IgG2 同种型，并识别免疫显性核蛋白氨基末端亚区内的主要 B 细胞表位。使用 mAb 5F11 作为固相第一抗体，β-D-半乳糖苷酶缀合 mAb 5E3 作为该蛋白质的第二抗体，我们建立了一种特异性 HCV 核心蛋白捕获 FEIA，能够检测低至 20 pg/ml 的 HCV 核心蛋白。重组HCV核心蛋白，经4.0%聚乙二醇、0.5 M NaOH、5% Triton X-100处理后，血清中可检测到HCV核心蛋白。肽抑制测定结果表明，该FEIA对HCV RNA阳性血清具有特异性，血清中检测到的HCV核心蛋白的量与HCV RNA的水平显着相关。 HCV核心蛋白的检出限为每滴度HCV RNA约10(4)/ml。使用该FEIA系统，慢性HCV感染患者中HCV核心蛋白的检出率为92.3%(70/76)。
A protein-capture fluorescence enzyme immunoassay (FEIA) was developed using monoclonal antibodies (mAbs) against recombinant hepatitis C virus (HCV) core protein, Four hybridoma cell lines (5E3, 5F11, 515S, 1080S) were established and characterized. These monoclonal antibodies (mAbs) each had IgG1 and IgG2 isotypes, and recognized major B cell epitopes within the immunodominant nucleoprotein amino terminal subregion, Using mAb 5F11 as the first antibody to the solid phase and beta-D-galactosidase-conjugated mAb 5E3 as the second antibody to the protein, we established a specific HCV core protein capturing FEIA capable of detecting as little as 20 pg/ml of recombinant HCV core protein, HCV core protein in serum was detectable after treatment with 4.0% polyethyleneglycol, 0.5 M NaOH, and 5% Triton X-100. The results of a peptide inhibition assay indicated that this FEIA is specific for HCV RNA positive sera, The quantity of HCV core protein detected in serum was significantly correlated to the level of HCV RNA. The detection limit for HCV core proteins was an HCV RNA per titer of similar to 10(4)/ml, Using this FEIA system, the detection ratio of HCV core protein in patients with chronic HCV infection was 92.3% (70/76).