Kaiser Laurent, Perrin Luc, Chapuis Bernard, Hadaya Karine, Kolarova Lenka, Deffernez Christelle, Huguet Saadia, Helg Claudine, Wunderli Werner
Division of Infectious Diseases. Division of Hematology. Division of Oncology, University Hospital of Geneva, Geneva, Switzerland.
J Clin Microbiol. 2002 Nov;40(11):4251-5. doi: 10.1128/JCM.40.11.4251-4255.2002.
Cytomegalovirus (CMV) DNA amplification assays in plasma have shown limited sensitivity compared to the detection of pp65 antigen in leukocytes. Our goal was to increase the sensitivity of a commercial CMV DNA PCR quantitative assay. After modification, the new assay was able to reproducibly detect 20 CMV DNA copies/ml of plasma. We compared this new ultrasensitive PCR assay with the standard PCR and the pp65 test for CMV detection and quantification in 22 consecutive allogeneic hematopoietic stem cell recipients. CMV infection or reactivation was detected in 84 of 319 (26%) samples by the ultrasensitive PCR assay compared to 38 of 319 (12%) samples by the pp65 assay (P < 0.01). All samples positive by the pp65 assay were positive by the ultrasensitive PCR, and CMV episodes were detected on average 4 days earlier and 7 days later than the first and the last pp65-positive test, respectively. In addition, during CMV episodes, the ultrasensitive assay identified positive samples that were inconsistently detected by the pp65 assay. The ultrasensitive assay was also much more sensitive than the standard PCR, with 26 versus 12% of CMV DNA-positive samples (P < 0.01). This assay improved the monitoring of CMV infection or reactivation in hematopoietic allogeneic stem cell recipients.
与检测白细胞中的pp65抗原相比，血浆中的巨细胞病毒（CMV）DNA扩增检测显示出有限的灵敏度。我们的目标是提高一种商用CMV DNA PCR定量检测的灵敏度。经过改进后，新检测方法能够可重复地检测出血浆中每毫升20个CMV DNA拷贝。我们将这种新的超灵敏PCR检测方法与标准PCR以及pp65检测方法进行了比较，用于对22例连续的异基因造血干细胞接受者进行CMV检测和定量分析。通过超灵敏PCR检测，在319份样本中的84份（26%）检测到CMV感染或再激活，而通过pp65检测在319份样本中的38份（12%）检测到（P<0.01）。所有通过pp65检测呈阳性的样本通过超灵敏PCR检测也呈阳性，并且CMV发作分别比第一次和最后一次pp65阳性检测平均提前4天和推迟7天被检测到。此外，在CMV发作期间，超灵敏检测方法识别出了pp65检测方法未一致检测到的阳性样本。超灵敏检测方法也比标准PCR灵敏得多，CMV DNA阳性样本分别为26%和12%（P<0.01）。该检测方法改善了对异基因造血干细胞接受者中CMV感染或再激活的监测。
