{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,19]],"date-time":"2026-09-19T10:33:08Z","timestamp":1789813988286,"version":"4.0.1"},"reference-count":24,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2018,3,9]],"date-time":"2018-03-09T00:00:00Z","timestamp":1520553600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"Yong Siew Yoon Research Grant"},{"name":"National University Cancer Institute, Singapore (NCIS)"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Head &amp; Neck"],"published-print":{"date-parts":[[2018,7]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Chromogenic Epstein\u2010Barr virus\u2010encoded RNA (EBER) in situ hybridization (EBER\u2010ISH) is the gold standard to detect Epstein\u2010Barr virus (EBV) but it is difficult to use in conjunction with immunohistochemistry (IHC). In this study, our purpose was to validate the sensitivity and specificity of RNAscope in detection of EBV infection in nasal epithelium and its stroma.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Fluorescence\u2010based RNAscope EBER\u2010ISH, <jats:italic>BRLF1<\/jats:italic>\u2010ISH, and lineage marker\u2010IHC were performed on archived formalin\u2010fixed paraffin\u2010embedded tissues from normal nasal cavity (n = 5), nasopharynx (n = 8), and nasopharyngeal carcinoma (NPC) specimens (n = 10).<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The EBERs were detected in 10 of 10 NPC samples but was absent in all normal tissues from the nasal cavity and nasopharynx. The EBERs were exclusively located in pan\u2010cytokeratin (pan\u2010CK)\u2010positive tumor epithelial cells but not in CD45\u2010positive leukocytes and vimentin\u2010positive stromal fibroblasts. The level of EBER expression varied in tumor cells within patient and between patients as well. Additionally, 5 of 10 patients had positive <jats:italic>BRLF<\/jats:italic>\u2010ISH.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>We developed a simple and reproducible method to simultaneously detect mRNA and protein in formalin\u2010fixed paraffin\u2010embedded tissues of NPC. As a single staining, traditional EBER continues to be useful; however, for interpretation of the phenotype of EBV\u2010infected cells, RNAscope is superior. Significantly, we showed that lytic EBV infection took place in NPC tumors.<\/jats:p><\/jats:sec>","DOI":"10.1002\/hed.25131","type":"journal-article","created":{"date-parts":[[2018,3,9]],"date-time":"2018-03-09T16:07:54Z","timestamp":1520611674000},"page":"1515-1523","source":"Crossref","is-referenced-by-count":16,"title":["Presence of lytic Epstein\u2010Barr virus infection in nasopharyngeal carcinoma"],"prefix":"10.1002","volume":"40","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2715-4790","authenticated-orcid":false,"given":"Fenggang","family":"Yu","sequence":"first","affiliation":[{"name":"Department of Otolaryngology, Yong Loo Lin School of Medicine National University of Singapore  Singapore"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanan","family":"Lu","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology, Yong Loo Lin School of Medicine National University of Singapore  Singapore"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fredrik","family":"Petersson","sequence":"additional","affiliation":[{"name":"Department of Pathology, Yong Loo Lin School of Medicine National University of Singapore  Singapore"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"De\u2010Yun","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology, Yong Loo Lin School of Medicine National University of Singapore  Singapore"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kwok Seng","family":"Loh","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology, Yong Loo Lin School of Medicine National University of Singapore  Singapore"},{"name":"Head and Neck Tumor Group, National Cancer Institute of Singapore, Head and Neck Surgery National University Health System (NUHS)  Singapore"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2018,3,9]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199509143331103"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/newbio244245a0"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/ijc.2910170102"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/path.1711650105"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1097\/00000478-198908000-00001"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1002\/ijc.2910530616"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/jmv.1890450114"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01677-16"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/0046-8177(93)90031-B"},{"key":"e_1_2_8_11_1","first-page":"1461","article-title":"Abundant expression of EBER1 small nuclear RNA in nasopharyngeal carcinoma. 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