使用新型小鼠单克隆抗体检测血清丙型肝炎病毒E1/E2抗原诊断丙型肝炎病毒感染
Diagnosis of Hepatitis C Virus Infection Using a Novel Mouse Monoclonal Antibody to Detect Serum HCV E1/E2 Antigens.
作者信息
Touni Iman, Tabll Ashraf A, Sobieh Shaimaa S, Hewedy Maha A, Abd Yasmine S El, Viazov Sergei
出版信息
Clin Lab. 2017 Apr 1;63(4):669-678. doi: 10.7754/Clin.Lab.2016.161124.
BACKGROUND
Hepatitis C virus (HCV) is a major cause of liver disease worldwide and in Egypt. The aim of this study was to detect HCV E1/E2 antigens using a novel mouse monoclonal antibody (mAb) designated (7G9) as a diagnostic and alternative approach for HCV detection.
METHODS
The detection of HCV-E1/E2 antigens in 138 patients positive for HCV infection tested by RT-PCR and 25 healthy individuals negative for HCV as control group was done by an optimized in-house ELISA and DotELISA (based on the molecular mimicry of E2 to immunoglobulins).
RESULTS
The mAb (7G9) was found to be IgM (heavy-chain)/kappa (light-chain) and characterization by western blot revealed two bands at 63 kDa for E2 and 31 kDa for E1. ELISA peptide mapping showed high reactivity with peptide derived from HCV E1 (a.a. 315 - 323) and low reactivity to peptides derived from HCV E2 (a.a. 517 - 531) and HCV E2 (a.a. 412 - 419). The mAb (7G9) showed no reactivity with HBV Ag, S. typhi or B. Abortus Ag proving high specificity. AUC for HCV-E1/E2 detection was 0.96 for all HCV patients with sensitivity 87% (119/137), specificity 88% (22/25) and efficiency 87%. The HCV-E1/E2 antigens detection by Dot-ELISA showed 76.8% sensitivity, 88% specificity and the efficiency of the assay was 78.5%. Furthermore, no correlation was found between serum HCV viral load and HCV E1/E2 antigens detection.
CONCLUSIONS
The ELISA and Dot-ELISA based on the monoclonal antibody (7G9) are reliable, rapid, easy and economic diagnostic assays for HCV infection.
背景
丙型肝炎病毒（HCV）是全球及埃及肝病的主要病因。本研究旨在使用一种名为（7G9）的新型小鼠单克隆抗体（mAb）检测HCV E1/E2抗原，作为HCV检测的诊断及替代方法。
方法
通过优化的内部酶联免疫吸附测定（ELISA）和斑点酶联免疫吸附测定（DotELISA）（基于E2与免疫球蛋白的分子模拟），对138例经逆转录聚合酶链反应（RT-PCR）检测HCV感染呈阳性的患者以及25例HCV呈阴性的健康个体作为对照组进行HCV-E1/E2抗原检测。
结果
发现单克隆抗体（7G9）为IgM（重链）/κ（轻链），蛋白质印迹法鉴定显示E2在63 kDa处有两条带，E1在31 kDa处有两条带。ELISA肽图谱显示与源自HCV E1（氨基酸315 - 323）的肽具有高反应性，而与源自HCV E2（氨基酸517 - 531）和HCV E2（氨基酸412 - 419）的肽反应性较低。单克隆抗体（7G9）与乙肝病毒表面抗原（HBV Ag）、伤寒沙门菌或流产布鲁氏菌抗原无反应，证明具有高特异性。所有HCV患者HCV-E1/E2检测的曲线下面积（AUC）为0.96，敏感性为87%（119/137），特异性为88%（22/25），效率为87%。斑点酶联免疫吸附测定（Dot-ELISA）检测HCV-E1/E2抗原的敏感性为76.8%，特异性为88%，该检测方法的效率为78.5%。此外，血清HCV病毒载量与HCV E1/E2抗原检测之间未发现相关性。
结论
基于单克隆抗体（7G9）的ELISA和DotELISA是用于HCV感染的可靠、快速、简便且经济的诊断检测方法。
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