Sandwich type ELISA and a fluorescent cytometric microbead assay for quantitative determination of hepatitis B virus X antigen level in human sera
Sandwich type ELISA and a fluorescent cytometric microbead assay for quantitative determination of hepatitis B virus X antigen level in human sera
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DOI：
10.1016/j.jim.2005.08.011
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发表时间：
2005-11-30
影响因子：
2.2
通讯作者：
Németh, P
中科院分区：
文献类型：
--
作者：
Pál, J;Pálinkás, L;Németh, P
乙型肝炎病毒 X 蛋白 (HBxAg) 是造成 HBV 感染严重并发症（包括原发性肝细胞癌）的原因。介绍了用于定量测定血清 Hbx-Ag 水平的夹心式 ELISA 和流式细胞术微珠测定法。我们之前开发了针对 HbxAg 上保守的表位的单克隆抗体家族，其特征在于不同的免疫组织化学和免疫血清学技术。抗体对的特殊选择为定量免疫分析的开发提供了高灵敏度和高特异性的工具。对从患有不同临床形式的 HBV 感染的患者收集的人血清（208 个样本）进行了测试。根据重组抗原和慢性肝炎患者血清的测量结果，夹心型 ELISA 的灵敏度范围为 4 至 2000 ng/ml。建立了进一步的流式细胞术微珠测定法，并与 ELISA 平行进行测试。这两种免疫血清学技术的定量结果具有很强的相关性，并且对临床样本具有高度的特异性和敏感性。 HBxAg ELISA 技术适用于常规临床实验室测量，我们的 HBxAg 微珠技术建议与其他标记物结合用于复杂的多参数测量。 (c) 2005 Elsevier B.V. 保留所有权利。
The hepatitis B virus X protein (HBxAg) is responsible for severe complications of HBV infections including primary hepatocellular carcinoma. A sandwich type ELISA and a flow cytometric microbead assay for quantitative determination of serum levels of Hbx-Ag are introduced. We have previously developed monoclonal antibody families against well-conserved epitopes on HbxAg, characterized by different immunohistochemical and immunoserological techniques. Special selection of the antibody pairs provided highly sensitive and highly specific tools for quantitative immunoassay development. The resulting assays were tested on human sera (208 samples) collected from patients suffering from different clinical forms of HBV infection. The sensitivity range of the sandwich type ELISA was between 4 and 2000 ng/ml as measured on both the recombinant antigen and the sera of chronic hepatitis patients. A further flow cytometric microbead assay was established and tested in parallel with the ELISA. The quantitative results of these two immunoserological techniques were in strongcorrelation and they were found to be highly specific and sensitive on clinical samples. The HBxAg ELISA technique is applicable for routine clinical laboratory measurements, and our HBxAg microbead technique is recommended for complex multiparametric measurements combined with other markers. (c) 2005 Elsevier B.V. All rights reserved.