Discordant detection of human cytomegalovirus DNA from peripheral blood mononuclear cells, granulocytes and plasma: correlation to viremia and HCMV infection
Discordant detection of human cytomegalovirus DNA from peripheral blood mononuclear cells, granulocytes and plasma: correlation to viremia and HCMV infection
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DOI：
10.1016/s1386-6532(98)00046-4
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发表时间：
1998-08-20
影响因子：
8.8
通讯作者：
Jahn, G
中科院分区：
文献类型：
--
作者：
Hamprecht, K;Steinmassl, M;Jahn, G
背景：目前关于免疫抑制患者中单一阳性白细胞亚型的HCMV感染的资料很少。大多数报告描述了HCMV共感染的细胞的骨髓单核细胞系，甚至T-和B-细胞群体。两种主要白细胞组分和血浆的阳性PCR结果与病毒血症和HCMV感染的相关性一般应有助于选择合适的HCMV DNA来源用于诊断目的。同时分离外周血单个核细胞定性检测白血病的诊断价值通过比较血液中的巢式PCR阳性结果与从白细胞或任何其他细胞中分离的病毒，研究设计：从75名临床怀疑原发性或复发性HCMV感染的免疫抑制患者的总共220份血液样品中制备PBMC、粒细胞和血浆。在35例主要接受骨髓或实体器官移植的患者中，白血病或血浆DNA血症的阳性结果与HCMV筛查和临床状态的数据相关。为了标准化，从每个白细胞群体的100 ng细胞DNA扩增HCMV IE外显子4 DNA。可以排除交叉污染。用苯酚/氯仿提取血浆中的DNA。结果：在30例HCMV阳性的白血病患者中，仅12例（40%）出现PBMC和粒细胞双重感染。然而，在大多数情况下（18例患者，60%），HCMV DNA检测仅在一个白细胞部分，无论是在PBMC或粒细胞。白血病与病毒血症无关。HCMV DNA。从血浆中扩增的细胞显示无细胞。未从血浆中分离出感染性病毒。结果表明，血浆和PBMC的nPCR检测HCMV感染的预测值较高，其预测值依次为血浆（0.92）> PBMC（0.83）>粒细胞（0.65）。然而，单个阳性白细胞组分的比率接近60%。因此，在40%的病例中，病毒性白细胞DNA血症似乎仅限于PBMC或粒细胞。出于诊断目的，应使用整个白细胞群体进行PCR分析。(C)1998 Elsevier Science B. V.保留所有权利。
Background: There exist only few data about the HCMV infection of single positive leukocyte subtypes in immunosuppressed patients. Most reports describe HCMV coinfection of cells of the myelomonocytic line or even T- and B-cell populations. Correlation of positive PCR findings from two major leukocyte fractions and plasma to viremia and HCMV infection in general should contribute to select suitable sources of HCMV DNA for diagnostic purposes.Objective: The diagnostic value of qualitative leukoDNAemia of simultaneously isolated peripheral blood mononuclear cells (PBMC), granulocytes as well as plasmaDNAemia was evaluated by comparing the positive results of nested PCR from blood with virus isolation either from leukocytes or from any other sources, with serology and the clinical status of immunosuppressed patients.Study design: PBMC, granulocytes and plasma were prepared of a total of 220 blood samples of 75 immunosuppressed patients with clinically suspected primary or recurrent HCMV infection. In a collective of 35 patients consisting mainly of recipients of marrow or solid organ transplants positive results of leuko- or plasmaDNAemia were correlated with data from HCMV screening and the clinical status. For standardization, HCMV IE Exon 4 DNA was amplified from 100 ng cellular DNA of each leukocyte population. Cross contamination can be excluded. DNA from plasma was extracted by phenol/chloroform. Using this experimental design, HCMV DNA was not detectable in PBMC, granulocytes and plasma of 23 healthy HCMV seropositive blood donors.Results: Leukocyte separation in a collective of 30 patients with positive leukoDNAemia revealed in only 12 cases (40%) double infection of PBMC and granulocytes. In the majority of cases (18 patients, 60%) however, HCMV DNA was detectable in only one leukocyte fraction, either in PBMC or granulocytes. LeukoDNAemia did not correlate to viremia. HCMV DNA. amplified from plasma was shown to be cell free. infectious virus from plasma was not isolated. The predictive value of qualitative nested PCR from blood to detect HCMV infection was high for plasma and decreased in the following sequence: plasma (0.92) > PBMC (0.83) > granulocytes (0.65).Conclusions: Qualitative nPCR from plasma and PBMC seems to be sufficient to detect (an ongoing) HCMV infection of immunosuppressed patients. However, the rate of single positive leukocyte fractions is similar to 60%. Therefore, viral leukoDNAemia in 40% of cases seems to be restricted to either PBMC or granulocytes. For diagnostic purposes the whole leukocyte population should be used for PCR analysis. (C) 1998 Elsevier Science B.V. All rights reserved.