Hadaya Karine, Wunderli Werner, Deffernez Christelle, Martin Pierre-Yves, Mentha Gilles, Binet Isabelle, Perrin Luc, Kaiser Laurent
Division of Infectious Diseases, University Hospitals of Geneva, 1211 Geneva 14, Switzerland.
J Clin Microbiol. 2003 Aug;41(8):3757-64. doi: 10.1128/JCM.41.8.3757-3764.2003.
Early and accurate monitoring of cytomegalovirus (CMV) infection in solid-organ transplant recipients is of major importance. We have assessed the potential benefit of an ultrasensitive plasma-based PCR assay for renal transplant recipients. The pp65 CMV antigen (pp65 Ag) assay using leukocytes was employed as a routine test for the monitoring of CMV in 23 transplant recipients. We compared the pp65 antigenemia with the CMV load quantified by an ultrasensitive PCR (US-PCR) with a limit of detection of 20 CMV DNA copies/ml of plasma. CMV infection was detected in 215 (67%) of 321 plasma samples by the US-PCR compared with 124 (39%) of 321 samples by the pp65 Ag assay. The US-PCR assay permitted the detection of CMV infection episodes following transplantation a median of 12 days earlier than the pp65 Ag assay. Moreover, during CMV infection episodes, DNA detection by the US-PCR was consistently positive, whereas false negative results were frequently observed with the pp65 Ag assay. We found a good correlation between the two assays, and the peak viral loads were significantly higher in patients with CMV-related complications (median, 5000 DNA copies/ml) than in those without symptoms (1160 DNA copies/ml) (P = 0.048). In addition, patients that did not require preemptive therapy based on the results of the pp65 assay had CMV loads significantly lower (median, 36 DNA copies/ml) than those that needed treatment (median, 4703 DNA copies/ml) (P < 0.001). These observations provided cutoff levels that could be applied in clinical practice. The ultrasensitive plasma-based PCR detected CMV infection episodes earlier and provided more consistent results than the pp65 Ag assay. This test could improve the monitoring of CMV infection or reactivation in renal transplant recipients.
对实体器官移植受者进行早期准确的巨细胞病毒（CMV）感染监测至关重要。我们评估了一种基于超敏血浆的PCR检测方法对肾移植受者的潜在益处。采用白细胞pp65 CMV抗原（pp65 Ag）检测法作为23例移植受者CMV监测的常规检测方法。我们将pp65抗原血症与通过超敏PCR（US-PCR）定量的CMV载量进行了比较，该检测方法的检测限为每毫升血浆20个CMV DNA拷贝。通过US-PCR在321份血浆样本中的215份（67%）检测到CMV感染，而通过pp65 Ag检测法在321份样本中的124份（39%）检测到CMV感染。US-PCR检测法比pp65 Ag检测法平均提前12天检测到移植后CMV感染发作。此外，在CMV感染发作期间，US-PCR检测DNA始终呈阳性，而pp65 Ag检测法经常出现假阴性结果。我们发现两种检测方法之间具有良好的相关性，并且CMV相关并发症患者的病毒载量峰值（中位数为5000个DNA拷贝/毫升）显著高于无症状患者（1160个DNA拷贝/毫升）（P = 0.048）。此外，根据pp65检测结果不需要进行抢先治疗的患者的CMV载量（中位数为36个DNA拷贝/毫升）显著低于需要治疗的患者（中位数为4703个DNA拷贝/毫升）（P < 0.001）。这些观察结果提供了可应用于临床实践的临界值。基于超敏血浆的PCR检测比pp65 Ag检测能更早地检测到CMV感染发作，并提供更一致的结果。该检测方法可改善肾移植受者CMV感染或再激活的监测。
