One-step multiplex real-time PCR assay to analyse the latency patterns of Epstein-Barr virus infection
One-step multiplex real-time PCR assay to analyse the latency patterns of Epstein-Barr virus infection
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DOI：
10.1016/j.jviromet.2007.08.012
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发表时间：
2008-01-01
影响因子：
3.1
通讯作者：
Kimura, Hiroshi
中科院分区：
文献类型：
--
作者：
Kubota, Naomi;Wada, Kaoru;Kimura, Hiroshi
EB病毒（EBV）建立了三种类型的病毒基因表达的潜伏感染。这些潜伏类型可以通过EBV核抗原（EBNA）1、EBNA 2、潜伏膜蛋白（LMP）1和LMP 2的表达模式来区分。EBV裂解周期由EBV立即早期BZLF1基因的转录启动，其可用于区分潜伏感染和裂解感染。本研究开发了一步多重实时PCR测定，以通过相对定量同时定量EBNA1、EBNA2、LMP1、LMP2和BZLF 1表达水平。为了验证该试验，在EBV阳性B、T和自然杀伤细胞系中进行了病毒基因转录的定量。由于其快速性、敏感性和特异性，这种新的检测方法可用于定量分析EBV感染的潜伏模式以及从潜伏期到裂解性病毒复制的转变。(C)2007 Elsevier B.V.保留所有权利。
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. (C) 2007 Elsevier B.V. All rights reserved.