Dual-Labeled Time-Resolved Immunofluorometric Assay for the Simultaneous Quantitative Detection of Hepatitis B Virus Antigens in Human Serum

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• 作者：Rong-Liang Liang ; Yun-Sen Yang ; Jian-Wei Zhou ; Tian-Cai Liu…
• 关键词：Hepatitis B virus surface antigen ; Hepatitis B e antigen ; Time ; resolved fluoroimmunoassay ; Dual ; label
• 刊名：Journal of Fluorescence
• 出版年：2017
• 出版时间：January 2017
• 年：2017
• 卷：27
• 期：1
• 页码：309-316
• 全文大小：
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Biomedicine, general; Biological and Medical Physics, Biophysics; Biotechnology; Biochemistry, general; Analytical Chemistry;
• 出版者：Springer US
• ISSN：1573-4994
• 卷排序：27

文摘

In this paper, a novel time-resolved fluorescence immunoassay (TRFIA) is described that allows the simultaneous quantitative detection of hepatitis B virus surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) in human serum to aid the diagnosis and monitoring of hepatitis B virus infection. The proposed method was developed based on a two-step sandwich immunoassay protocol in which monoclonal antibodies against HBsAg and HBeAg were co-coated in 96 microtitration wells, then tracer polyclonal antibodies against HBsAg labeled with samarium and tracer monoclonal antibodies against HBeAg labeled with europium chelates were used for detection. The detection range was 0.1–150 IU/mL for HBsAg and 0.5–160 PEIU/mL for HBeAg, and the detection limits were 0.03 IU/L and 0.09 PEIU/ml, respectively. The intra- and inter-assay coefficients of variation were below 8 % for both virus antigens. The dilution linearity and accuracy of the assay were satisfactory. No statistically significant differences were observed in sensitivity or specificity for the serum samples between the dual-label TRFIA and a commercial single-label TRFIA. These results demonstrate that an effective, reliable and convenient HBsAg/HBeAg dual-label TRFIA was successfully developed that may be clinically applicable for blood screening to monitor the course of hepatitis B virus infection and predict treatment responses.