title: One-step multiplex real-time PCR assay to analyse the latency patterns of Epstein-Barr virus infection.
abstract: Epstein-Barr virus (EBV) establishes a latent infection with three types of viral gene expression. These latency types can be distinguished by the expression patterns of EBV nuclear antigen (EBNA)1, EBNA2, latent membrane protein (LMP)1, and LMP2. The EBV lytic cycle is initiated by the transcription of the EBV immediate early BZLF1 gene, which can be used to distinguish between a latent and a lytic infection. In this study, a one-step multiplex real-time PCR assay was developed to quantify the EBNA1, EBNA2, LMP1, LMP2, and BZLF1 expression levels simultaneously by relative quantification. To validate this assay, the quantitation of viral gene transcription was performed in EBV-positive B, T, and natural killer cell lines. Because of its rapidity, sensitivity, and specificity, this new assay can be used for quantitative analyses of the latency patterns of EBV infection and the switch from latency to lytic viral replication.
authors.affiliation: Department of Virology, Nagoya University Graduate School of Medicine, 65 Tsuruma-cho, Showa-ku, Nagoya 466-8550, Japan.
authors.forename: Naomi
authors.initials: N
authors.lastname: Kubota
authors.forename: Kaoru
authors.initials: K
authors.lastname: Wada
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Ito
authors.forename: Yoshie
authors.initials: Y
authors.lastname: Shimoyama
authors.forename: Shigeo
authors.initials: S
authors.lastname: Nakamura
authors.forename: Yukihiro
authors.initials: Y
authors.lastname: Nishiyama
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kimura
mesh_terms: D001402:B-Lymphocytes / Q000166:cytology / Q000821:virology; D002460:Cell Line; D020031:Epstein-Barr Virus Infections / Q000821:virology*; D015870:Gene Expression; D005814:Genes, Viral; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D006801:Humans; D007694:Killer Cells, Natural / Q000166:cytology / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D013601:T-Lymphocytes / Q000166:cytology / Q000821:virology; D014764:Viral Proteins / Q000032:analysis* / Q000235:genetics; D017735:Virus Latency
chemical_list: D014764:Viral Proteins