{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,5]],"date-time":"2026-03-05T18:18:11Z","timestamp":1772734691572,"version":"3.50.1"},"reference-count":27,"publisher":"American Society for Microbiology","issue":"5","license":[{"start":{"date-parts":[[2009,5,1]],"date-time":"2009-05-01T00:00:00Z","timestamp":1241136000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Clin Vaccine Immunol"],"published-print":{"date-parts":[[2009,5]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>Undifferentiated nasopharyngeal carcinoma (NPC; WHO type III) is 100% associated with Epstein-Barr virus (EBV) infection and the fourth most prevalent cancer in Indonesian males. Therapy failure is high, since most patients come to the hospital at an advanced stage of disease. Screening for early-stage NPC is needed. Here, a simple and economical two-step enzyme-linked immunosorbent assay (ELISA) system is proposed for diagnosing NPC in high-risk populations, employing the peptide-based immunoglobulin A (IgA) EBNA1 plus viral capsid antigen p18 ELISA as an initial screening test and the IgA early antigen (EA) ELISA using a different set of EBV antigens as a confirmation test. A total of 151 NPC patients and 199 regional healthy EBV carriers were used to evaluate the two-step ELISA approach. Routinely, EBV IgG immunoblotting is used as a standard confirmation test. The sensitivity and specificity for diagnosing NPC by the two-step ELISA approach increased from 85.4% to 96.7% and 90.1% to 98%, respectively, with positive predictive values and negative predictive values increasing from 78.7 and 93.9% to 97.3 and 97.5%, respectively, relative to the immunoblotting confirmation system. On discrepant samples, additional testing was done by EBV DNA load quantification in blood. Results showed that 5\/11 discrepant NPC samples with an elevated IgA EA ELISA also had elevated an EBV DNA load in the circulation (range, 3,200 to 25,820 copies\/ml). Therefore, the IgA EA ELISA is proposed as a confirmation test in first-line NPC serological screening studies. This two-step EBV ELISA system provides a standardized approach for NPC screening and may be used in combination with dried blood sampling in future field studies for identification of early-stage NPC in high-risk regions.<\/jats:p>","DOI":"10.1128\/cvi.00425-08","type":"journal-article","created":{"date-parts":[[2009,3,26]],"date-time":"2009-03-26T01:36:10Z","timestamp":1238031370000},"page":"706-711","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":30,"title":["Two-Step Epstein-Barr Virus Immunoglobulin A Enzyme-Linked Immunosorbent Assay System for Serological Screening and Confirmation of Nasopharyngeal Carcinoma"],"prefix":"10.1128","volume":"16","author":[{"given":"Dewi K.","family":"Paramita","sequence":"first","affiliation":[{"name":"Department of Histology and Cell Biology, Gadjah Mada University, Yogyakarta, Indonesia"},{"name":"Department of Pathology, Vrije Universiteit Medical Center, Amsterdam, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jajah","family":"Fachiroh","sequence":"additional","affiliation":[{"name":"Department of Histology and Cell Biology, Gadjah Mada University, Yogyakarta, Indonesia"},{"name":"Department of Pathology, Vrije Universiteit Medical Center, Amsterdam, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sofia M.","family":"Haryana","sequence":"additional","affiliation":[{"name":"Department of Histology and Cell Biology, Gadjah Mada University, Yogyakarta, Indonesia"},{"name":"Department of Pathology, Vrije Universiteit Medical Center, Amsterdam, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jaap M.","family":"Middeldorp","sequence":"additional","affiliation":[{"name":"Department of Histology and Cell Biology, Gadjah Mada University, Yogyakarta, Indonesia"},{"name":"Department of Pathology, Vrije Universiteit Medical Center, Amsterdam, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"crossref","first-page":"706","DOI":"10.1002\/ijc.11130","volume":"105","year":"2003","unstructured":"Chan, K. H., Y. L. Gu, F. Ng, P. S. Ng, W. H. Seto, J. S. Sham, D. Chua, W. Wei, Y. L. Chen, W. Luk, Y. S. Zong, and M. H. Ng. 2003. EBV specific antibody-based and DNA-based assays in serologic diagnosis of nasopharyngeal carcinoma. Int. J. Cancer105:706-709.","journal-title":"Int. J. Cancer"},{"key":"e_1_3_2_3_2","doi-asserted-by":"crossref","first-page":"1877","DOI":"10.1056\/NEJMoa011610","volume":"345","year":"2001","unstructured":"Chien, Y. C., J. Y. Chen, M. Y. Liu, H. I. Yang, M. M. Hsu, J. Y. Chen, and C. S. Yang. 2001. Serologic markers of Epstein-Barr virus infection and nasopharyngeal carcinoma in Taiwanese men. N. Engl. J. Med.345:1877-1882.","journal-title":"N. Engl. J. Med."},{"key":"e_1_3_2_4_2","first-page":"301","volume":"149","year":"2001","unstructured":"Crowther, J. R. 2001. Validation of diagnostic test for infectious disease. Methods Mol. Biol.149:301-346.","journal-title":"Methods Mol. Biol."},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.39.9.3164-3170.2001"},{"key":"e_1_3_2_6_2","doi-asserted-by":"crossref","first-page":"1806","DOI":"10.1002\/ijc.22857","volume":"121","year":"2007","unstructured":"de Sanjose, S., R. Bosch, T. Schouten, S. A. Verkuijlen, A. Nieters, L. Foretova, M. Maynadie, P. L. Cocco, A. Staines, N. Becker, P. Brennan, Y. Benavente, P. Boffetta, C. J. Meijer, and J. M. Middeldorp. 2007. Epstein-Barr virus infection and risk of lymphoma: immunoblot analysis of antibody responses against EBV-related proteins in large series of lymphoma subjects and matched controls. Int. J. Cancer121:1806-1812.","journal-title":"Int. J. Cancer"},{"key":"e_1_3_2_7_2","doi-asserted-by":"crossref","first-page":"176","DOI":"10.1002\/ijc.2910420206","volume":"42","year":"1988","unstructured":"de Vathaire, F., H. Sancho-Garnier, H. de The, C. Pieddeloup, G. Schwab, J. H. Ho, R. Ellousz, C. Michaeu, M. Cammoun, Y. Cachin, and G. de The. 1988. Prognostic value of EBV markers in the clinical management of nasopharyngeal carcinoma (NPC): a multicenter follow-up study. Int. J. Cancer42:176-181.","journal-title":"Int. J. Cancer"},{"key":"e_1_3_2_8_2","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1086\/421245","volume":"190","year":"2004","unstructured":"Fachiroh, J., T. Schouten, B. Hariwiyanto, D. K. Paramita, A. Harijadi, S. M. Haryana, M. H. Ng, and J. M. Middeldorp. 2004. Molecular diversity of Epstein-Barr virus IgG and IgA antibody responses in nasopharyngeal carcinoma: a comparison of Indonesian, Chinese, and European subjects. J. Infect. Dis.190:53-62.","journal-title":"J. Infect. Dis."},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.44.4.1459-1467.2006"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.01368-07"},{"key":"e_1_3_2_11_2","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1016\/S0166-0934(00)00301-3","volume":"93","year":"2001","unstructured":"Gartner, B. C., J. M. Fischinger, K. Roemer, M. Mak, B. Fleurent, and N. Mueller-Lantzsch. 2001. Evaluation of recombinant line blot for diagnosis of Epstein-Barr virus compared with ELISA, using immunofluorescence as reference method. J. Virol. Methods93:89-96.","journal-title":"J. Virol. Methods"},{"key":"e_1_3_2_12_2","doi-asserted-by":"crossref","first-page":"78","DOI":"10.1128\/CDLI.10.1.78-82.2003","volume":"10","year":"2003","unstructured":"Gartner, B. C., R. D. Hess, D. Bandt, A. Kruse, A. Rethwilm, K. Roemer, and N. Mueller-Lantzsch. 2003. Evaluation of four commercially available Epstein-Barr virus enzyme immunoassays with an immunofluorescence assay as the reference method. Clin. Diagn. Lab. Immunol.10:78-82.","journal-title":"Clin. Diagn. Lab. Immunol."},{"key":"e_1_3_2_13_2","first-page":"225","volume":"44","year":"1970","unstructured":"Henle, W., G. Henle, H. C. Ho, P. Burtin, Y. Cachin, P. Clifford, A. De Schryver, G. de The, V. Diehl, and G. Klein. 1970. Antibodies to Epstein Barr virus in nasopharyngeal carcinoma, other head and neck neoplasm group, and control group. J. Int. Cancer Inst.44:225-231.","journal-title":"J. Int. Cancer Inst."},{"key":"e_1_3_2_14_2","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1002\/ijc.2910170102","volume":"17","year":"1976","unstructured":"Henle, G., and W. Henle. 1976. Epstein-Barr Virus specific IgA serum antibodies as an outstanding feature of nasopharyngeal carcinoma. Int. J. Cancer17:1-7.","journal-title":"Int. J. Cancer"},{"key":"e_1_3_2_15_2","first-page":"55","volume":"34","year":"1976","unstructured":"Ho, H. C., M. H. Ng, H. C. Kwan, and J. C. Chau. 1976. Epstein-Barr virus specific IgA and IgG serum antibodies in nasopharyngeal carcinoma. Br. J. Cancer34:55-60.","journal-title":"Br. J. Cancer"},{"key":"e_1_3_2_16_2","doi-asserted-by":"crossref","first-page":"623","DOI":"10.1038\/sj.bjc.6603609","volume":"96","year":"2007","unstructured":"Ji, M. F., D. K. Wang, Y. L. Yu, Y. Q. Guo, J. S. Liang, W. M. Cheng, Y. S. Zong, K. H. Chan, S. P. Ng, W. I. Wei, D. T. T. Chua, J. S. T. Sham, and M. H. Ng. 2007. Sustained elevation of Epstein-Barr virus antibody levels preceding clinical onset of nasopharyngeal carcinoma. Br. J. Cancer96:623-630.","journal-title":"Br. J. Cancer"},{"key":"e_1_3_2_17_2","doi-asserted-by":"crossref","first-page":"593","DOI":"10.1002\/jmv.20310","volume":"75","year":"2005","unstructured":"Karray, H., W. Ayadi, L. Fki, A. Hammami, J. Daoud, M. M. Drira, M. Frikha, R. Jlidi, and J. M. Middeldorp. 2005. Comparison of three different serological techniques for primary diagnosis and monitoring of nasopharyngeal carcinoma in two age groups from Tunisia. J. Med. Virol.75:593-602.","journal-title":"J. Med. Virol."},{"key":"e_1_3_2_18_2","doi-asserted-by":"crossref","first-page":"631","DOI":"10.1200\/JCO.2003.06.158","volume":"21","year":"2003","unstructured":"Lin, J. C., J. S. Jan, C. Y. Hsu, W. M. Liang, R. S. Jiang, and W. Y. Wang. 2003. Phase III study of concurrent chemoradiotherapy versus radiotherapy alone for advanced nasopharyngeal carcinoma: positive effect on overall and progression-free survival. J. Clin. Oncol.21:631-637.","journal-title":"J. Clin. Oncol."},{"key":"e_1_3_2_19_2","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1001\/archotol.132.1.82","volume":"132","year":"2006","unstructured":"Loh, K. S., B. C. Goh, J. Lu, W. S. Hsieh, and L. Tan. 2006. Familial nasopharyngeal carcinoma in a cohort of 200 patients. Arch. Otolaryngol. Head Neck Surg.132:82-85.","journal-title":"Arch. Otolaryngol. Head Neck Surg."},{"key":"e_1_3_2_20_2","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1016\/0166-0934(88)90060-2","volume":"21","year":"1988","unstructured":"Middeldorp, J. M., and P. Herbrink. 1988. Epstein-Barr virus specific marker molecules for early diagnosis of infectious mononucleosis. J. Virol. Methods21:133-146.","journal-title":"J. Virol. Methods"},{"key":"e_1_3_2_21_2","doi-asserted-by":"crossref","first-page":"998","DOI":"10.1002\/ijc.20672","volume":"113","year":"2005","unstructured":"Ng, W. T., T. K. Yau, R. W. Yung, W. M. Sze, A. H. Tsang, A. L. Law, and A. W. Lee. 2005. Screening for family members of patients with nasopharyngeal carcinoma. Int. J. Cancer113:998-1001.","journal-title":"Int. J. Cancer"},{"key":"e_1_3_2_22_2","first-page":"250","volume":"25","year":"2006","unstructured":"Ng, M. H., K. H. Chan, S. P. Ng, and Y. S. Zong. 2006. Epstein-Barr virus serology in early detection and screening of nasopharyngeal carcinoma. Chinese J. Cancer25:250-256.","journal-title":"Chinese J. Cancer"},{"key":"e_1_3_2_23_2","doi-asserted-by":"crossref","first-page":"1710","DOI":"10.1002\/jmv.20987","volume":"79","year":"2007","unstructured":"Paramita, D. K., J. Fachiroh, W. T. Artama, E. van Benthem, S. M. Haryana, and J. M. Middeldorp. 2007. Native early antigen of Epstein-Barr virus, a promising antigen for diagnosis of nasopharyngeal carcinoma. J. Med. Virol.79:1710-1721.","journal-title":"J. Med. Virol."},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.43.7.3066-3073.2005"},{"key":"e_1_3_2_25_2","doi-asserted-by":"crossref","first-page":"1040","DOI":"10.1002\/hed.20466","volume":"28","year":"2006","unstructured":"Stevens, S. J. C., S. A. W. M. Verkuijlen, M. C. Zwaan, and J. M. Middeldorp. 2006. Epstein-Barr virus (EBV) serology, but not EBV DNA load, for predicting distant metastases in a juvenile Caucasian nasopharyngeal carcinoma (NPC) patient without clinical response upon EBV lytic induction therapy. Head Neck28:1040-1045.","journal-title":"Head Neck"},{"key":"e_1_3_2_26_2","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1002\/jmv.1890400215","volume":"40","year":"1993","unstructured":"van Grunsven, W. M. J., A. Nabbe, and J. M. Middeldorp. 1993. Identification and molecular characterization of two diagnostically relevant marker proteins of the Epstein-Barr virus capsid antigen complex. J. Med. Virol.40:161-169.","journal-title":"J. Med. Virol."},{"key":"e_1_3_2_27_2","first-page":"47","year":"1997","unstructured":"WHO International Agency for Cancer Research. 1997. Epstein-Barr virus, p. 47-373. IARC Monographs on the Evaluation of Carcinogenic Risks in Humans, publ. 70. IARC Press, Lyon, France.","journal-title":"Epstein-Barr virus"},{"key":"e_1_3_2_28_2","doi-asserted-by":"crossref","first-page":"599","DOI":"10.1002\/ijc.2910310511","volume":"31","year":"1983","unstructured":"Zeng, Y., C. H. Gong, M. G. Jan, Z. Fun, L. G. Zhang, and H. Y. Li. 1983. Detection of Epstein-Barr virus IgA\/EA antibody for diagnosis of nasopharyngeal carcinoma by immunoautoradiography. Int. J. Cancer31:599-601.","journal-title":"Int. J. Cancer"}],"container-title":["Clinical and Vaccine Immunology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/CVI.00425-08","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/CVI.00425-08","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,7,29]],"date-time":"2021-07-29T15:55:07Z","timestamp":1627574107000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/CVI.00425-08"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2009,5]]},"references-count":27,"journal-issue":{"issue":"5","published-print":{"date-parts":[[2009,5]]}},"alternative-id":["10.1128\/CVI.00425-08"],"URL":"https:\/\/doi.org\/10.1128\/cvi.00425-08","relation":{},"ISSN":["1556-6811","1556-679X"],"issn-type":[{"value":"1556-6811","type":"print"},{"value":"1556-679X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2009,5]]},"assertion":[{"value":"2008-11-15","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2009-03-12","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2020-12-18","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}