Hakim H, Gibson C, Pan J, Srivastava K, Gu Z, Bankowski M J, Hayden R T
Department of Infectious Diseases, St. Jude Children's Research Hospital, 332 North Lauderdale Street, Memphis, TN 38105, USA.
J Clin Microbiol. 2007 Jul;45(7):2151-5. doi: 10.1128/JCM.02308-06. Epub 2007 May 9.
Epstein-Barr virus (EBV) infection is associated with a broad spectrum of disease. While quantification of EBV nucleic acid in the peripheral blood has been demonstrated to be useful for diagnosis and patient care, the optimal sample type and reporting format for such testing remain uncertain. Using quantitative real-time PCR (QRT-PCR), we evaluated EBV in whole blood (WB), peripheral blood mononuclear cells (PBMC), and plasma in 249 samples from 122 patients. In WB and PBMC, results were reported both in viral copies/ml and in copies/microg of total DNA. Trendings of quantitative values over time among the different sample types were compared. The sensitivities of QRT-PCR using WB and that using PBMC did not differ significantly (P = 0.33), and both were more sensitive than plasma alone (P < 0.0001). EBV viral load results from WB and PBMC paired sample types also showed a significant correlation (P < 0.05), as did results reported in copies/ml and copies/microg DNA for both WB and PBMC (R2 > 0.93). EBV viral loads detected using WB and PBMC trended very closely for the few patients who had multiple positive samples available for analysis. WB and PBMC show comparable sensitivities and a close quantitative correlation when assayed for EBV by QRT-PCR. The close correlation between copies/ml and copies/microg DNA also suggests that normalization to cell number or genomic DNA in cellular specimens may not be necessary.
爱泼斯坦-巴尔病毒(EBV)感染与多种疾病相关。虽然外周血中EBV核酸的定量已被证明对诊断和患者护理有用,但此类检测的最佳样本类型和报告格式仍不确定。我们使用定量实时PCR(QRT-PCR)对122例患者的249份样本中的全血(WB)、外周血单个核细胞(PBMC)和血浆中的EBV进行了评估。在全血和外周血单个核细胞中,结果以病毒拷贝数/毫升和每微克总DNA中的拷贝数两种方式报告。比较了不同样本类型中定量值随时间的变化趋势。使用全血的QRT-PCR和使用外周血单个核细胞的QRT-PCR的敏感性无显著差异(P = 0.33),且两者均比单独检测血浆更敏感(P < 0.0001)。全血和外周血单个核细胞配对样本类型的EBV病毒载量结果也显示出显著相关性(P < 0.05),全血和外周血单个核细胞以拷贝数/毫升和每微克DNA报告的结果也是如此(R2 > 0.93)。对于少数有多个阳性样本可供分析的患者,使用全血和外周血单个核细胞检测到的EBV病毒载量趋势非常接近。通过QRT-PCR检测EBV时,全血和外周血单个核细胞显示出可比的敏感性和紧密的定量相关性。拷贝数/毫升和每微克DNA之间的紧密相关性还表明,对细胞样本中的细胞数量或基因组DNA进行标准化可能没有必要。