Neville J A, Prescott L E, Bhattacherjee V, Adams N, Pike I, Rodgers B, El-Zayadi A, Hamid S, Dusheiko G M, Saeed A A, Haydon G H, Simmonds P
Department of Medical Microbiology, University of Edinburgh, United Kingdom.
J Clin Microbiol. 1997 Dec;35(12):3062-70. doi: 10.1128/jcm.35.12.3062-3070.1997.
Assays that detect antibody to hepatitis C virus (HCV) are used to screen blood donors and patients with hepatitis. Current enzyme-linked immunosorbent assay (ELISA)-based methods are invariably based upon antigens from expressed recombinant proteins or oligopeptides from HCV type 1. Some HCV antigens used in screening assays are coded by regions of the HCV genome that show extensive variability; therefore, HCV type 1-based assays may be less effective for the detection of antibody elicited by infection with other genotypes. In this study, we have measured antibody reactivity of sera from 110 hepatitis C patients infected with type 1b, 3a, or 4a to genotype-specific and cross-reactive epitopes present in recombinant proteins from HCV genotypes 1b (core, NS3, and NS5), 3a (NS3, NS5), and 4a (core, NS3), corresponding to those used in current third-generation screening ELISAs. By comparing the serological reactivities of sera to type-homologous and type-heterologous antigens, we detected a significant type-specific component to the reactivity to NS3 (61 to 77% of the total reactivity) and NS5 (60% of the total reactivity). Furthermore, despite the similarities in the amino acid sequences of the core antigens of type 1b and type 4a, we also found significantly greater reactivity to type-homologous antigens, with approximately 25% of reactivity being type specific. These findings are consistent with previous findings of fivefold weaker reactivity of sera from HCV type 2- and HCV type 3-infected blood donors in the currently used third-generation ELISAs and suggest that these assays are suboptimal for screening populations in which the predominant genotype is not type 1.
检测丙型肝炎病毒（HCV）抗体的检测方法用于筛查献血者和肝炎患者。目前基于酶联免疫吸附测定（ELISA）的方法总是基于来自1型HCV表达的重组蛋白或寡肽的抗原。筛查检测中使用的一些HCV抗原由HCV基因组中显示出广泛变异性的区域编码；因此，基于1型HCV的检测方法对于检测由其他基因型感染引发的抗体可能效果较差。在本研究中，我们测定了110例感染1b型、3a型或4a型的丙型肝炎患者血清对来自HCV 1b型（核心、NS3和NS5）、3a型（NS3、NS5）和4a型（核心、NS3）重组蛋白中基因型特异性和交叉反应性表位的抗体反应性，这些重组蛋白对应于当前第三代筛查ELISA中使用的那些。通过比较血清对同型和异型抗原的血清学反应性，我们检测到对NS3（占总反应性的61%至77%）和NS5（占总反应性的60%）的反应性中有显著的型特异性成分。此外，尽管1b型和4a型核心抗原的氨基酸序列相似，但我们也发现对同型抗原具有显著更高的反应性，约25%的反应性是型特异性的。这些发现与先前的发现一致，即在目前使用的第三代ELISA中，来自2型HCV和3型HCV感染献血者的血清反应性弱五倍，并表明这些检测方法对于筛查主要基因型不是1型的人群并非最佳。
