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Hiroshi Kimura, Kanae Miyake, Yohei Yamauchi, Kana Nishiyama, Seiko Iwata, Keiji Iwatsuki, Kensei Gotoh, Seiji Kojima, Yoshinori Ito, Yukihiro Nishiyama, Identification of Epstein-Barr Virus (EBV)–Infected Lymphocyte Subtypes by Flow Cytometric In Situ Hybridization in EBV-Associated Lymphoproliferative Diseases, The Journal of Infectious Diseases, Volume 200, Issue 7, 1 October 2009, Pages 1078–1087, https://doi.org/10.1086/605610
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Abstract
To diagnose Epstein-Barr virus (EBV)–associated diseases and to explore the pathogenesis of EBV infection, not only must the EBV load be measured, but EBV-infected cells must also be identified. We established a novel flow cytometric in situ hybridization assay to detect EBV+ suspension cells using a peptide nucleic acid probe specific for EBV-encoded small RNA (EBER). By enhancing fluorescence and photostability, we successfully stained EBER and surface antigens on the same cells. In 3 patients with hydroa vacciniforme–like lymphoproliferative disease, we demonstrated that 1.7%–25.9% of peripheral lymphocytes were infected with EBV and specifically identified these lymphocytes as CD3+CD4−CD8− γδ T cell receptor–positive T cells. The results indicate that this novel and noninvasive assay is a direct and reliable method of characterizing EBV-infected lymphocytes that can be used not only to diagnose EBV infection but also to clarify the pathogenesis of EBV-associated diseases