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Epidemiology of Epstein–Barr virus, cytomegalovirus, and kaposi's sarcoma‐associated herpesvirus infections in peripheral blood leukocytes revealed by a multiplex PCR assay
Author(s) -
Nishiwaki Morie,
Fujimuro Masahiro,
Teishikata Yasuhiro,
Inoue Hisanori,
Sasajima Hitoshi,
Nakaso Kazuhiro,
Nakashima Kenji,
Sadanari Hidetaka,
Yamamoto Tomohiro,
Fujiwara Yoshie,
Ogawa Naoki,
Yokosawa Hideyoshi
Publication year - 2006
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.20748
Subject(s) - virology , primary effusion lymphoma , biology , multiplex polymerase chain reaction , multiplex , virus , cytomegalovirus , polymerase chain reaction , herpesviridae , gammaherpesvirinae , human cytomegalovirus , epstein–barr virus , kaposi's sarcoma associated herpesvirus , viral disease , gene , bioinformatics , biochemistry
A multiplex polymerase chain reaction (PCR) has been developed for the simultaneous detection of Epstein–Barr virus (EBV), cytomegalovirus (CMV), and Kaposi's sarcoma‐associated herpesvirus (KSHV) in a clinical sample. Primers of multiplex PCR were designed to amplify specific regions of the EBV EBNA1, CMV IE2, and KSHV LANA genes. This multiplex PCR assay was found to have detection sensitivities of 1–10 copies of purified viral DNA cloned into the plasmid. To assess diagnostic and pre‐clinical applications with this method, we utilized KSHV‐positive primary effusion lymphoma (PEL) cells, EBV‐positive Burkitt's lymphoma cells, CMV‐infected fibroblast cells, and clinically prepared peripheral blood leukocytes (PBLs) that had been infected with viruses. We found that this multiplex PCR assay has high sensitivity and specificity for simultaneous detection of EBV, CMV, and KSHV genomes in a single amplification from a clinical material. Using this multiplex PCR assay, we investigated the prevalence of EBV, CMV, and KSHV in PBL samples from normal Japanese randomly selected. KSHV, EBV, and CMV genomes were detected in samples from 2 (0.2%), 377 (39.5%), and 27 (2.8%) of the 953 blood donors, respectively. Interestingly, both EBV and CMV genomes were detected in samples from all KSHV‐positive donors. J. Med. Virol. 78:1635–1642, 2006. © 2006 Wiley‐Liss, Inc.