Department of Biomedicine, University of Basel, Switzerland.
J Med Virol. 2011 Dec;83(12):2143-50. doi: 10.1002/jmv.22232.
Surveillance of cytomegalovirus (CMV) replication in transplant patients is crucial for the success of transplantation. To compare a CMV pp65 antigenemia (pp65Ag) and a quantitative real-time PCR targeting the CMV-UL111a (UL111aPCR), all whole blood samples taken between July 2008 and October 2009 were identified which had been analyzed prospectively by both assays in parallel. Discordant results were re-analyzed using a published CMV duplex PCR targeting regions UL55 and UL123exon4. Of 720 samples from 81 transplant patients, CMV replication was detected in 244 specimens (34%) by the UL111aPCR (median, 1,019 geq/ml), compared to 113 (16%) detected by the pp65Ag (median, 2/250,000 leukocytes). Concordant UL111aPCR/pp65Ag results were obtained in 561 (78%) samples, being positive in 99 (14%), and negative in 462 (64%). As a rule of thumb, 1 pp65Ag-positive cell per 250,000 leukocytes corresponded to 1,000 geq/ml CMV DNA of whole blood. Discordant results were found in 159 samples (22%), being UL111aPCR-positive/pp65Ag-negative in 145 (91%; median, 650 geq/ml), or UL111aPCR-negative/pp65Ag-positive in 14 (9%; median, 1/250,000 cells). Using the duplex PCR targeting the CMV UL55 and the UL123-exon4 genes, 131 of 139 (94%) discordant UL111aPCR-positives (median UL111aPCR, 639 geq/ml; median UL55PCR, 715 geq/ml; median UL123PCR, 1,103 geq/ml) were confirmed. Of 14 discordant pp65Ag-positives, duplex PCR was also negative in 8, and of low copy number in 6. Thus, CMV UL111aPCR provides more sensitive quantitation of CMV replication than pp65Ag, however, discordant results can occur at very low viral loads.
对移植患者的巨细胞病毒（CMV）复制进行监测对于移植的成功至关重要。为了比较 CMV pp65 抗原血症（pp65Ag）和针对 CMV-UL111a 的定量实时 PCR（UL111aPCR），对 2008 年 7 月至 2009 年 10 月期间所有前瞻性平行分析的全血样本进行了鉴定。使用针对区域 UL55 和 UL123exon4 的已发表的 CMV 双重 PCR 对不一致的结果进行了重新分析。在 81 名移植患者的 720 份样本中，通过 UL111aPCR（中位数，1,019geq/ml）检测到 244 份（34%）CMV 复制，而通过 pp65Ag（中位数，2/250,000 白细胞）检测到 113 份（16%）。561 份（78%）样本获得了一致的 UL111aPCR/pp65Ag 结果，99 份（14%）为阳性，462 份（64%）为阴性。作为经验法则，每 250,000 个白细胞中有 1 个 pp65Ag 阳性细胞对应于全血中 1,000geq/ml 的 CMV DNA。在 159 份（22%）样本中发现了不一致的结果，145 份（91%；中位数，650geq/ml）为 UL111aPCR 阳性/pp65Ag 阴性，14 份（9%）为 UL111aPCR 阴性/pp65Ag 阳性（中位数，1/250,000 细胞）。使用针对 CMV UL55 和 UL123-exon4 基因的双重 PCR，131 份（94%）不一致的 UL111aPCR 阳性结果得到了确认（中位数 UL111aPCR，639geq/ml；中位数 UL55PCR，715geq/ml；中位数 UL123PCR，1,103geq/ml）。在 14 份不一致的 pp65Ag 阳性样本中，双重 PCR 也有 8 份为阴性，6 份为低拷贝数。因此，CMV UL111aPCR 比 pp65Ag 更能灵敏地定量 CMV 复制，但在病毒载量非常低的情况下可能会出现不一致的结果。
