Epidemiology of Epstein–Barr virus, cytomegalovirus, and kaposi's sarcoma-associated herpesvirus infections in peripheral blood leukocytes revealed by a multiplex PCR assay
Article first published online: 24 OCT 2006
Copyright © 2006 Wiley-Liss, Inc.
Journal of Medical Virology
Volume 78, Issue 12, pages 1635–1642, December 2006
How to Cite
Nishiwaki, M., Fujimuro, M., Teishikata, Y., Inoue, H., Sasajima, H., Nakaso, K., Nakashima, K., Sadanari, H., Yamamoto, T., Fujiwara, Y., Ogawa, N. and Yokosawa, H. (2006), Epidemiology of Epstein–Barr virus, cytomegalovirus, and kaposi's sarcoma-associated herpesvirus infections in peripheral blood leukocytes revealed by a multiplex PCR assay. J. Med. Virol., 78: 1635–1642. doi: 10.1002/jmv.20748
- Issue published online: 24 OCT 2006
- Article first published online: 24 OCT 2006
- Manuscript Accepted: 29 AUG 2006
- Ministry of Education, Culture, Sports, Science and Technology of Japan
- Takeda Science Foundation
- Japan Health Sciences Foundation and the New Energy and Industrial Technology Development Organization (NEDO) of Japan
- multiplex polymerase chain reaction;
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.