Freymuth F, Gennetay E, Petitjean J, Eugene G, Hurault de Ligny B, Ryckelynck J P, Legoff C, Hazera P, Bazin C
Virus Laboratory, University Hospital Caen, France.
J Clin Microbiol. 1994 Jun;32(6):1614-8. doi: 10.1128/jcm.32.6.1614-1618.1994.
A nested PCR was used for the detection of human cytomegalovirus (HCMV) DNA in plasma. The presence of HCMV DNA and its correlation to pp65 leukocytic antigenemia were investigated with 299 blood samples from 45 organ transplant recipients and 63 AIDS patients. Of the 53 samples positive by nested PCR, 52 (98%) were also positive for leukocytic antigenemia and 23 had high levels of antigenemia (> 50 positive cells per 2 x 10(5) leukocytes). Of the 246 samples negative in PCR, only 3 (1.2%) had highly positive antigenemia. For 15 patients having a high antigenemia level in the course of their disease, consecutive blood samples were studied and also assessed for viremia in culture. The extent to which HCMV DNA, detected by PCR, was present in plasma correlated with increased levels of HCMV leukocytic antigenemia for six of the eight AIDS patients and for all the organ transplant recipients. Positivity for HCMV DNA in PCR and for viremia in cell culture was usually restricted to the highest antigenemia levels. From a total of 69 blood samples, PCR and culture gave positive results, respectively, for 17 of 32 samples (53%) and 14 of 32 samples (43%) from transplant recipients and for 15 of 37 samples (40%) and 9 of 37 samples (24%) from AIDS patients. Our findings have shown a strong correlation between high levels of leukocytic antigenemia and HCMV DNA in plasma. The detection of HCMV DNA in plasma by this nested PCR can prove HCMV dissemination in blood, but it lacks the rapidity and simplicity of the leukocytic pp65 antigenemia procedure.
采用巢式聚合酶链反应(PCR)检测血浆中的人巨细胞病毒(HCMV)DNA。对45例器官移植受者和63例艾滋病患者的299份血样进行检测,以研究HCMV DNA的存在情况及其与pp65白细胞抗原血症的相关性。在巢式PCR检测呈阳性的53份样本中,52份(98%)白细胞抗原血症也呈阳性,其中23份抗原血症水平较高(每2×10⁵白细胞中阳性细胞>50个)。在PCR检测呈阴性的246份样本中,仅有3份(1.2%)白细胞抗原血症呈强阳性。对15例在病程中抗原血症水平较高的患者,采集连续血样并进行病毒血症培养评估。在8例艾滋病患者中的6例以及所有器官移植受者中,PCR检测到的血浆中HCMV DNA的含量与HCMV白细胞抗原血症水平升高相关。PCR检测HCMV DNA呈阳性以及细胞培养检测病毒血症呈阳性通常仅限于最高抗原血症水平。在总共69份血样中,器官移植受者的32份样本中分别有17份(53%)PCR检测呈阳性、14份(43%)培养检测呈阳性;艾滋病患者的37份样本中分别有15份(40%)PCR检测呈阳性、9份(24%)培养检测呈阳性。我们的研究结果表明白细胞抗原血症高水平与血浆中HCMV DNA之间存在密切相关性。通过这种巢式PCR检测血浆中的HCMV DNA可证实HCMV在血液中的播散,但它缺乏白细胞pp65抗原血症检测方法的快速性和简便性。