Systematic screening for novel, serologically reactive Hepatitis E Virus epitopes.
Systematic screening for novel, serologically reactive Hepatitis E Virus epitopes.
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DOI：
10.1186/1743-422x-9-28
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发表时间：
2012-01-23
期刊：
影响因子：
4.8
通讯作者：
Baiker A
中科院分区：
文献类型：
--
作者：
Osterman A;Vizoso Pinto MG;Haase R;Nitschko H;Jäger S;Sander M;Motz M;Mohn U;Baiker A
美国国立卫生研究院将戊型肝炎归类为一种新兴疾病，因为戊型肝炎病毒（HEV）是发展中国家急性肝炎的主要原因。有趣的是，在工业化国家，越来越多的散发性HEV感染病例被描述为来自家畜的人畜共患病。尽管该病原体在临床病毒学中的相关性越来越大，但商业抗体检测主要基于HEV开放阅读框（ORF）2和ORF 3的片段。然而，最大的ORF 1（多）蛋白并不是当前测试格式的一部分。从合成的全长HEV基因型1 cDNA文库中，我们构建了由15个相应的HEV ORF结构域组成的完整HEV基因文库。在变性条件下细菌表达和纯化九种重组HEV蛋白后，以微阵列形式使用来自具有已知感染状态的患者的55份血清进行血清分析实验。SPSS软件通过进行受试者操作者特征、逻辑回归和相关性分析，评估了这9个ORF结构域与7种商业HEV抗原（基因型1和3）相比的抗原潜力。用我们的方法生产的用于血清分析实验的HEV抗原表现出与商业抗原相同的质量和特征。血清分析实验检测到Y，V和X结构域作为ORF 1-抗原，具有潜在的可比诊断意义，作为ORF 2和ORF 3的明确表位。然而，如生物信息学分析所揭示的，在诊断测试中没有实现灵敏度或特异性的明显额外增加。此外，我们发现潜在的跨膜蛋白ORF 3的C-末端结构域负责IgG和IgM血清反应性。数据表明，同源ORF 2抗原可能存在基因型特异性血清反应性。ORF 1表位的诊断价值可能不一定提高灵敏度和特异性，但扩大了现有测试系统的整体质量。ORF 2和ORF 3抗原仍然常用于诊断测定，并且可能具有血清学区分基因型1和3感染的潜力。我们的系统方法是一个合适的方法来调查他们的血清抗原性戊型肝炎病毒域。天然病毒结构域的表位筛选可能是开发新的血清学测试组分的优选工具。
The National Institutes of Health classified Hepatitis E as an emerging disease since Hepatitis E Virus (HEV) is the major cause of acute hepatitis in developing countries. Interestingly, an increasing number of sporadic cases of HEV infections are described in industrialized countries as zoonosis from domestic livestock. Despite the increasing relevance of this pathogen in clinical virology, commercial antibody assays are mainly based on fragments of HEV open reading frame (ORF) 2 and ORF3. The largest ORF1 (poly-)protein, however, is not part of current testing formats. From a synthesized full length HEV genotype 1 cDNA-bank we constructed a complete HEV gene library consisting of 15 respective HEV ORF domains. After bacterial expression and purification of nine recombinant HEV proteins under denaturating conditions serum profiling experiments using 55 sera from patients with known infection status were performed in microarray format. SPSS software assessed the antigenic potential of these nine ORF domains in comparison to seven commercial HEV antigens (genotype 1 and 3) by performing receiver operator characteristics, logistic regression and correlation analysis. HEV antigens produced with our method for serum profiling experiments exhibit the same quality and characteristics as commercial antigens. Serum profiling experiments detected Y, V and X domains as ORF1-antigens with potentially comparable diagnostic significance as the well established epitopes of ORF2 and ORF3. However no obvious additional increase in sensitivity or specificity was achieved in diagnostic testing as revealed by bioinformatic analysis. Additionally we found that the C-terminal domain of the potential transmembrane protein ORF3 is responsible for IgG and IgM seroreactivity. Data suggest that there might be a genotype specific seroreactivity of homologous ORF2-antigens. The diagnostic value of identified ORF1 epitopes might not necessarily improve sensitivity and specificity, but broaden the overall quality of existing test systems. ORF2 and ORF3-antigens are still commonly used in diagnostic assays and possibly hold the potential to serologically differentiate between genotype 1 and 3 infections. Our systematic approach is a suitable method to investigate HEV domains for their serologic antigenicity. Epitope screening of native viral domains could be a preferable tool in developing new serologic test components.