Wagner H J, Jabs W, Smets F, Wessel M, Fischer L, Offner G, Kirchner H, Bucsky P
Department of Pediatrics, Medical University of Lübeck, Germany.
Klin Padiatr. 2000 Jul-Aug;212(4):206-10. doi: 10.1055/s-2000-9678.
Measurement of Epstein-Barr virus (EBV) load is useful in peripheral blood for detecting primary and reactivated EBV-infections especially in immunosuppressed patients being at high risk for developing posttransplant lymphoproliferative disorder. For quantification of EBV DNA in peripheral blood of patients two real time polymerase chain reaction (RQ-PCR) assays were developed detecting sequences specific for the BAM HI-W and BAM HI-K region of EBV. In order to determine the optimal material of peripheral blood for RQ PCR analysis, DNA preparations of whole blood, peripheral blood mononuclear cells (PBMC) and B cells from 11 healthy, EBV-seropositive individuals were analysed in parallel and compared with regard to efficiency and sensitivity. While in whole blood preparations inhibitors of RQ PCR were detected influencing sensitivity, analysis of B cells being most sensitive is limited by being too labour intensive. In contrast, analysis of DNA preparations of PBMCs is sensitive enough to frequently detect EBV-specific sequences in all individuals tested and the preparation of PBMCs itself needs only a reasonable time. Thus, longitudinal monitoring of EBV load in peripheral blood of patients is possible by RQ-PCR, the optimal material for analysis being PBMCs.
测量爱泼斯坦-巴尔病毒（EBV）载量在外周血中对于检测原发性和再激活的EBV感染很有用，特别是在有发生移植后淋巴细胞增生性疾病高风险的免疫抑制患者中。为了定量患者外周血中的EBV DNA，开发了两种实时聚合酶链反应（RQ-PCR）检测方法，用于检测EBV的BAM HI-W和BAM HI-K区域的特异性序列。为了确定用于RQ-PCR分析的外周血的最佳材料，对11名健康的、EBV血清学阳性个体的全血、外周血单个核细胞（PBMC）和B细胞的DNA制备物进行了平行分析，并在效率和灵敏度方面进行了比较。虽然在全血制备物中检测到了影响灵敏度的RQ-PCR抑制剂，但对B细胞的分析最敏感，但由于劳动强度太大而受到限制。相比之下，PBMC的DNA制备物分析足够灵敏，能够在所有测试个体中频繁检测到EBV特异性序列，而且PBMC本身的制备只需要合理的时间。因此，通过RQ-PCR可以对外周血中患者的EBV载量进行纵向监测，分析的最佳材料是PBMC。
