title: Epidemiology of Epstein-Barr virus, cytomegalovirus, and Kaposi's sarcoma-associated herpesvirus infections in peripheral blood leukocytes revealed by a multiplex PCR assay.
abstract: 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.
authors.affiliation: GeneticLab Co., Ltd., Chuo-ku, Sapporo, Japan.
authors.forename: Morie
authors.initials: M
authors.lastname: Nishiwaki
authors.forename: Masahiro
authors.initials: M
authors.lastname: Fujimuro
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Teishikata
authors.forename: Hisanori
authors.initials: H
authors.lastname: Inoue
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Sasajima
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Nakaso
authors.forename: Kenji
authors.initials: K
authors.lastname: Nakashima
authors.forename: Hidetaka
authors.initials: H
authors.lastname: Sadanari
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Yamamoto
authors.forename: Yoshie
authors.initials: Y
authors.lastname: Fujiwara
authors.forename: Naoki
authors.initials: N
authors.lastname: Ogawa
authors.forename: Hideyoshi
authors.initials: H
authors.lastname: Yokosawa
mesh_terms: D001782:Blood Donors; D002460:Cell Line; D003587:Cytomegalovirus / Q000235:genetics / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000175:diagnosis / Q000453:epidemiology / Q000821:virology; D004279:DNA, Viral / Q000032:analysis; D020031:Epstein-Barr Virus Infections / Q000175:diagnosis / Q000453:epidemiology / Q000821:virology; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000453:epidemiology* / Q000821:virology; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000302:isolation & purification*; D019288:Herpesvirus 8, Human / Q000235:genetics / Q000302:isolation & purification*; D007962:Leukocytes / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015995:Prevalence; D012514:Sarcoma, Kaposi / Q000175:diagnosis / Q000453:epidemiology / Q000821:virology; D012680:Sensitivity and Specificity
chemical_list: D004279:DNA, Viral