SS Virologia Molecolare, SC Virologia e Microbiologia, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy.
J Clin Virol. 2012 Dec;55(4):317-22. doi: 10.1016/j.jcv.2012.08.017. Epub 2012 Sep 16.
Consensus human cytomegalovirus (HCMV) DNA cut-off values for preemptive therapy in transplant recipients have not yet been defined, mainly due to the lack of real-time PCR standardization.
(i) To compare the kinetics of HCMV DNA in transplanted patients using an in-house real-time PCR assay (Pavia assay) and the new Abbott RealTime CMV assay; (ii) to verify assay concordance in the identification of patients eligible for preemptive treatment and (iii) standardize results with international units (IUs) using the WHO International HCMV DNA Standard as a reference.
The kinetics of HCMV disseminated infection was retrospectively evaluated in 513 stored whole blood samples from 37 transplanted patients enrolled in randomized prospective studies designed for the clinical validation of HCMV DNA cut-off values. Conversion factors of HCMV DNA copy number to WHO international units (IUs) were determined.
Among the 513 samples, 352 (68.6%) were concordant positive, 42 (8.1%) concordant negative and 119 (23.1%) discordant. All discordant samples resulted positive by the Abbott RealTime CMV assay and negative by the Pavia assay, showing higher sensitivity for the Abbott RealTime CMV assay. A significant correlation was observed between concordant positive samples (r=0.89). HCMV DNAemia determined by each assay showed overlapping kinetics. Expression of results as IU/ml provided the best results in preemptive treatment simulation.
HCMV DNAemia cut-off values determined using our in-house assay and expressed as IU/ml appear valid for use in commercial assays as well as other potential in-house assays.
目前尚未确定移植受者抢先治疗的共识人巨细胞病毒（HCMV）DNA 截止值，主要是因为缺乏实时聚合酶链反应（PCR）标准化。
（i）比较使用内部实时 PCR 检测（帕维亚检测）和新的雅培实时 CMV 检测的移植患者 HCMV DNA 的动力学；（ii）验证在识别有资格进行抢先治疗的患者方面检测的一致性；（iii）使用世界卫生组织（WHO）国际 HCMV DNA 标准作为参考，将结果标准化为国际单位（IU）。
对纳入设计用于临床验证 HCMV DNA 截止值的随机前瞻性研究的 37 名移植患者的 513 份存储全血样本进行回顾性评估，以评估 HCMV 传播感染的动力学。确定 HCMV DNA 拷贝数转换为 WHO 国际单位（IU）的换算系数。
在 513 份样本中，352 份（68.6%）检测结果一致为阳性，42 份（8.1%）检测结果一致为阴性，119 份（23.1%）检测结果不一致。所有不一致的样本均通过雅培实时 CMV 检测呈阳性，而通过帕维亚检测呈阴性，表明雅培实时 CMV 检测的敏感性更高。一致阳性样本之间存在显著相关性（r=0.89）。两种检测方法确定的 HCMV 血症动力学具有重叠性。以 IU/ml 表示结果的表达在抢先治疗模拟中提供了最佳结果。
使用我们内部检测确定的 HCMV DNA 截止值，并以 IU/ml 表示，似乎可用于商业检测以及其他潜在的内部检测。
