Single-assay combination of Epstein-Barr virus (EBV) EBNA1-and viral capsid antigen-p18-derived synthetic peptides for measuring anti-EBV immunoglobulin G (IgG) and IgA antibody levels in sera from nasopharyngeal carcinoma patients: Options for field screening
Single-assay combination of Epstein-Barr virus (EBV) EBNA1-and viral capsid antigen-p18-derived synthetic peptides for measuring anti-EBV immunoglobulin G (IgG) and IgA antibody levels in sera from nasopharyngeal carcinoma patients: Options for field screening
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DOI：
10.1128/jcm.44.4.1459-1467.2006
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发表时间：
2006-04-01
影响因子：
9.4
通讯作者：
Middeldorp, JM
中科院分区：
文献类型：
--
作者：
Fachiroh, J;Paramita, DK;Middeldorp, JM
评价免疫球蛋白A（伊加）抗体对各种EB病毒（EBV）抗原复合物的反应，通常涉及多种血清学检测，对于鼻咽癌（NPC）的早期诊断具有重要意义。通过合成EBNA 1和病毒衣壳抗原（VCA）-p18的两个免疫优势表位的合成肽，建立了一种检测鼻咽癌患者EB病毒IgG和伊加抗体的一步法夹心酶联免疫吸附试验（EBVIgG/伊加ELISA）。血清来自印度尼西亚和中国的健康供体（n = 367）、非NPC头颈癌患者（n = 43）和活检证实的NPC患者（n = 296）。与健康EBV携带者相比，在NPC患者中观察到EBY IgG在450 nm处的光密度值更高，但大的重叠限制了其用于NPC诊断。单独使用EBNA 1或VCA-p18肽伊加ELISA正确鉴定了88.5%和79.8%的印度尼西亚NPC患者，特异性为80.1%和70.9%，而结合两种肽的单孔包被分别产生了90.1%和85.4%的灵敏度和特异性值。EBNA 1 + VCA伊加ELISA联合检测的阳性和阴性预测值（PPV和NPV）分别为78.7%和93.9%。在印度尼西亚组中，EBV伊加反应水平与NPC肿瘤大小、淋巴结受累、转移分期、性别和年龄组无关。在中国组中，单肽检测的灵敏度/特异性值为86.2/92.0%（EBNA 1伊加）和84.1/90.3%（VCA-p18伊加），VCA + EBNA 1伊加ELISA联合检测的灵敏度/特异性值为95.1/90.6%，EBV伊加ELISA联合检测的PPV和NPV分别为95.6%和89.3%。几乎所有的NPC患者有异常的抗-EBV IgG多样性模式所确定的免疫印迹分析。另一方面，EBV伊加ELISA阳性的健康EBV携带者显示正常的IgG多样性模式。通过使用EBVIgG免疫印迹多样性作为伊加ELISA阳性样本的确认检测，鼻咽癌诊断的灵敏度和特异性分别提高到98%和99.2%，在印尼鼻咽癌样本。建议使用这些综合方法进行血清流行病学筛查研究。
Assessment of immunoglobulin A (IgA) antibody responses to various Epstein-Barr virus (EBV) antigen complexes, usually involving multiple serological assays, is important for the early diagnosis of nasopharyngeal carcinoma (NPC). Through combination of two synthetic peptides representing immunodominant epitopes of EBNA1 and viral capsid antigen (VCA)-p18 we developed a one-step sandwich enzyme-linked immunosorbent assay (ELISA) for the specific detection of EBV reactive IgG and IgA antibodies in NPC patients (EBV IgG/IgA ELISA). Sera were obtained from healthy donors (n = 367), non-NPC head and neck cancer patients (n = 43), and biopsy-proven NPC patients (n = 296) of Indonesian and Chinese origin. Higher values of optical density at 450 nm for EBY IgG were observed in NPC patients compared to the healthy EBV carriers, but the large overlap limits its use for NPC diagnosis. Using either EBNA1 or VCA-p18 peptides alone IgA ELISA correctly identified 88.5% and 79.8% of Indonesian NPC patients, with specificities of 80.1% and 70.9%, whereas combined single-well coating with both peptides yielded sensitivity and specificity values of 90.1 and 85.4%, respectively. The positive and negative predictive values (PPV and NPV, respectively) for the combined EBNA1 plus VCA EBV IgA ELISA were 78.7% and 93.9%, respectively. In the Indonesia panel, the level of EBV IgA reactivity was not associated with NPC tumor size, lymph node involvement, and metastasis stage, sex, and age group. In the China panel the sensitivity/specificity values were 86.2/92.0% (EBNA1 IgA) and 84.1/90.3% (VCA-p18 IgA) for single-peptide assays and 95.1/90.6% for the combined VCA plus EBNA1 IgA ELISA, with a PPV and an NPV for the combined EBV IgA ELISA of 95.6 and 89.3%, respectively. Virtually all NPC patients had abnormal anti-EBV IgG diversity patterns as determined by immunoblot analysis. On the other hand, healthy EBV carriers with positive EBV IgA ELISA result showed normal IgG diversity patterns. By using EBV IgG immunoblot diversity as confirmation assay for EBV IgA ELISA-positive samples, the sensitivity and specificity for NPC diagnosis increased to 98% and 99.2%, respectively, in the Indonesian NPC samples. The use of these combined methods for seroepidemiological screening studies is proposed.