CHU de Rouen, Laboratoire de Virologie, F-76000 Rouen, France.
CHU de Rouen, Laboratoire de Virologie, F-76000 Rouen, France.
J Clin Virol. 2019 Apr;113:39-44. doi: 10.1016/j.jcv.2019.03.001. Epub 2019 Mar 2.
Measurement of cytomegalovirus (CMV) and Epstein-Barr virus (EBV) viral loads is commonly used to monitor posttransplant patients. Two new systems (eMAG/eSTREAM and Versant/kPCR) have been recently commercialized.
To evaluate the performance of four systems to quantify CMV and EBV in whole blood.
Three extraction and real-time PCR amplification systems: m2000SP/RT (M2000), eMAG/eSTREAM (EMAG), and Versant/kPCR (KPCR) were compared with our routine system Qiasymphony/RGQ (QS/RGQ). The 4 systems were tested using 3 dilutions in triplicate according to the WHO international standard (WHO-IS) for intra-assay reproducibility; 56 whole blood samples (24 patients, 4 follow-ups) for CMV and 45 samples (27 patients, 3 follow-ups) for EBV.
For CMV, the mean of the WHO-IS (expected value: 4.7 Log IU/ml) was: QS/RGQ=4.84, M2000=4.61, EMAG=4.33, and KPCR=4.79. One patient (10 samples) presented a major underquantification by QS/RGQ. Of the 46 remaining samples, 41 were quantified with QS/RGQ, 43 with M2000, 33 with EMAG and 24 with KPCR. For EBV, the mean of the WHO-IS was: QS/RGQ=4.70, M2000=4.61, EMAG=4.62, and KPCR=4.57. Among the 45 samples, 43 were quantified with QS/RGQ, 39 with M2000, 40 with EMAG and 32 with KPCR.
The results obtained with the WHO-IS were very good. The results of patients' samples were well correlated with the announced sensitivity of each system. The elevated threshold of the KPCR CMV assay may be problematic for the follow-up of highly immunocompromised patients who require early introduction of treatment.
巨细胞病毒 (CMV) 和 Epstein-Barr 病毒 (EBV) 病毒载量的测量常用于监测移植后患者。两个新系统（eMAG/eSTREAM 和 Versant/kPCR）最近已商业化。
评估四种系统定量检测全血中 CMV 和 EBV 的性能。
三种提取和实时 PCR 扩增系统：m2000SP/RT (M2000)、eMAG/eSTREAM (EMAG) 和 Versant/kPCR (KPCR) 与我们的常规系统 Qiasymphony/RGQ (QS/RGQ) 进行了比较。根据世界卫生组织国际标准 (WHO-IS) 进行了三次重复稀释试验，以评估各系统的批内可重复性；使用 56 份全血样本（24 例患者，4 次随访）进行 CMV 检测，使用 45 份样本（27 例患者，3 次随访）进行 EBV 检测。
对于 CMV，WHO-IS 的平均值（预期值：4.7 Log IU/ml）为：QS/RGQ=4.84，M2000=4.61，EMAG=4.33，KPCR=4.79。一名患者（10 份样本）的 QS/RGQ 检测结果明显偏低。在其余 46 份样本中，41 份样本可被 QS/RGQ 定量检测，43 份样本可被 M2000 定量检测，33 份样本可被 EMAG 定量检测，24 份样本可被 KPCR 定量检测。对于 EBV，WHO-IS 的平均值为：QS/RGQ=4.70，M2000=4.61，EMAG=4.62，KPCR=4.57。在 45 份样本中，43 份样本可被 QS/RGQ 定量检测，39 份样本可被 M2000 定量检测，40 份样本可被 EMAG 定量检测，32 份样本可被 KPCR 定量检测。
使用 WHO-IS 获得的结果非常好。患者样本的结果与每个系统公布的灵敏度高度相关。KPCR CMV 检测的高阈值可能会给需要早期引入治疗的高度免疫抑制患者的后续治疗带来问题。
