Shi Hongbo, Xie Li, Shi Honglin, Yan Li, Duan Zhongping
Beijing Artificial Liver Treatment and Training Centre, Beijing Youan Hospital, Affiliated Hospital of Capital Medical University, Beijing, China.
Hybridoma (Larchmt). 2012 Feb;31(1):54-9. doi: 10.1089/hyb.2011.0076.
Developing assays for detecting HCV antigens could be beneficial because viral proteins appear earlier than antibodies and are more stable than RNA in the serum. Monoclonal antibody was prepared by immunization and cell fusion. Subclass, specificity, and efficiency of monoclonal antibodies were determined by ELISA. Epitope specificity of monoclonal antibodies was analyzed by ELISA additivity test. HCV antigen in serum of hepatitis patients was examined by double monoclonal antibody sandwich ELISA. Five hybridoma cell lines were screened and named HCV(1), HCV(2), HCV(3), HCV(4), and HCV(5). These five monoclonal antibodies had high specificity and efficiency. The additivity test showed that HCV(2), HCV(4), and HCV(5) recognized different epitopes, which can be matched in ELISA. Of 173 anti-HCV positive patients, 37 (21.4%) were positive for HCV antigen. Of 1498 anti-HCV negative patients, 10 (0.67%) were positive for HCV antigen. Fifty normal controls were negative for HCV antigen. HCV antigen detection had moderate agreement and correlation with HCV RNA detection (kappa=0.577, p<0.01; r=0.59, p<0.01). This result indicates that the monoclonal antibody against HCV NS(3) may be a potential diagnostic reagent, which would provide a foundation for developing a sandwich ELISA of HCV antigen detection.
开发用于检测丙型肝炎病毒（HCV）抗原的检测方法可能是有益的，因为病毒蛋白比抗体出现得更早，并且在血清中比RNA更稳定。通过免疫和细胞融合制备单克隆抗体。通过酶联免疫吸附测定（ELISA）确定单克隆抗体的亚类、特异性和效率。通过ELISA加和试验分析单克隆抗体的表位特异性。采用双单克隆抗体夹心ELISA检测肝炎患者血清中的HCV抗原。筛选出5株杂交瘤细胞系，分别命名为HCV(1)、HCV(2)、HCV(3)、HCV(4)和HCV(5)。这5种单克隆抗体具有较高的特异性和效率。加和试验表明，HCV(2)、HCV(4)和HCV(5)识别不同的表位，可在ELISA中相互匹配。在173例抗HCV阳性患者中，37例（21.4%）HCV抗原阳性。在1498例抗HCV阴性患者中，10例（0.67%）HCV抗原阳性。50例正常对照HCV抗原均为阴性。HCV抗原检测与HCV RNA检测具有中度一致性和相关性（kappa=0.577，p<0.01；r=0.59，p<0.01）。这一结果表明，抗HCV NS(3)单克隆抗体可能是一种潜在的诊断试剂，为开发HCV抗原夹心ELISA检测奠定基础。
