Veal N, Payan C, Fray D, Sarol L, Blanchet O, Kouyoumdjian S, Lunel F
Laboratoire de Virologie, Centre Hospitalier Universitaire Angers, France.
J Clin Microbiol. 1996 Dec;34(12):3097-100. doi: 10.1128/jcm.34.12.3097-3100.1996.
We compared conventional cytomegalovirus (CMV) isolation, rapid viral culture, a CMV pp65 antigenemia assay, and a novel CMV DNA hybrid capture system (HCS). A total of 309 blood samples from individuals in different risk groups were assessed by at least two of the methods mentioned above. Leukocytes were recovered either after centrifugation in Leucosep tubes containing 1.080 Ficoll for pp65 assay or after simple differential lysis steps for DNA detection. HCS was based on DNA hybridization with a CMV RNA probe and its capture by antibodies to DNA-RNA hybrids. The CMV pp65 lower matrix protein was detected by fluorescence with c10-c11 monoclonal antibody in formalin-fixed leukocytes. Concordant results were observed for 92.9, 78.3, and 82.7% of the patients when comparing (i) viral culture and the pp65 antigenemia assay, (ii) viral culture and HCS, and (iii) the pp65 antigenemia assay and HCS, respectively. Discordant results were observed between a positive HCS result and negative culture and/or pp65 results. These results were associated with relatively low DNA levels (< 20 pg/10(6) cells) and positive viruria. In conclusion, the pp65 antigenemia assay is a rapid and reliable method of detecting CMV and is preferable to culture, but the Murex HCS appears to be more sensitive for CMV detection.
我们比较了传统的巨细胞病毒（CMV）分离法、快速病毒培养法、CMV pp65抗原血症检测法以及一种新型的CMV DNA杂交捕获系统（HCS）。采用上述至少两种方法对来自不同风险组的309份血液样本进行了评估。对于pp65检测，在含有1.080 Ficoll的Leucosep管中离心后回收白细胞；对于DNA检测，经过简单的差异裂解步骤后回收白细胞。HCS基于与CMV RNA探针的DNA杂交以及通过针对DNA-RNA杂交体的抗体进行捕获。在福尔马林固定的白细胞中，用c10-c11单克隆抗体通过荧光检测CMV pp65低基质蛋白。在比较（i）病毒培养和pp65抗原血症检测、（ii）病毒培养和HCS、（iii）pp65抗原血症检测和HCS时，分别有92.9%、78.3%和82.7%的患者观察到结果一致。在HCS结果为阳性而培养和/或pp65结果为阴性之间观察到了不一致的结果。这些结果与相对较低的DNA水平（<20 pg/10⁶细胞）和病毒尿阳性有关。总之，pp65抗原血症检测是一种快速可靠的检测CMV的方法，优于培养法，但Murex HCS在检测CMV方面似乎更敏感。
