Semiquantitative PCR analysis of Epstein‐Barr virus DNA in clinical samples of patients with EBV‐associated diseases
Semiquantitative PCR analysis of Epstein‐Barr virus DNA in clinical samples of patients with EBV‐associated diseases
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EBV 相关疾病患者临床样本中 Epstein-Barr 病毒 DNA 的半定量 PCR 分析
DOI：
10.1002/jmv.2040
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发表时间：
2001
影响因子：
12.7
通讯作者：
P. Wutzler
中科院分区：
文献类型：
--
作者：
A. Meerbach;B. Gruhn;R. Egerer;U. Reischl;F. Zintl;P. Wutzler
原发性和再激活 Epstein-Barr 病毒 (EBV) 感染的实验室诊断基于免疫功能正常患者的血清学方法。然而，在免疫功能低下的患者中，血清学数据难以解释，并且通常与临床数据不相关。为了找到诊断 EBV 相关疾病的有用且实用的标记物，建立了聚合酶链反应 (PCR) 检测方法来半定量检测 EBV 序列。该方法基于巢式 PCR，使用病毒衣壳抗原 p23 区域的引物和终点稀释。该方法对 39 名不同疾病患者的 68 份血浆样本、68 份外周血单核细胞样本和 5 份脑脊液样本进行了评估 EBV 基因组拷贝数。来自传染性单核细胞增多症患者的样本作为活动性 EBV 感染的阳性对照。在 5 名传染性单核细胞增多症患者中，在 105 个外周血单核细胞中检测到 EBV 基因组的高拷贝数，范围在 1,000-40,000 个拷贝之间。相比之下，19 名潜伏感染者的样本要么显示低拷贝数（105 个外周血单核细胞中有 10-100 个），要么 EBV PCR 呈阴性。在七名没有任何症状的肾移植患者中观察到了类似的结果。 EBV DNA 半定量检测的实用价值在三名骨髓移植受者中得到了证实。其中两人患上了淋巴组织增生性疾病，与血浆（分别为 16,000 和 50,000 拷贝/毫升）和外周血单核细胞（105 个外周血单核细胞中分别有 100,000 和 650 万拷贝）中极高的 EBV DNA 相关。 1 例成功抗病毒治疗后，血浆和外周血单核细胞中的高 EBV 载量显着降低。第三位骨髓移植受者出现了 EBV 诱导的横贯性脊髓炎，外周血单核细胞和 EBV 阳性脑脊液样本中 EBV 基因组拷贝数增加。经过抗病毒和免疫联合治疗后，EBV 基因组拷贝数下降，患者完全康复。这些数据表明 EBV 基因组的半定量检测与临床发现之间具有良好的相关性。推荐该方法用于诊断移植后患者的 EBV 相关疾病，以及监测治疗反应。 J. Med。病毒。 65:348–357, 2001。© 2001 Wiley-Liss, Inc.
The laboratory diagnosis of primary and reactivated Epstein‐Barr virus (EBV) infection is based on serologic methods in immunocompetent patients. However, in immunocompromised patients, serologic data are difficult to interpret and do not often correlate with clinical data. In order to find a useful and practical marker for diagnosis of EBV‐related diseases, a polymerase chain reaction (PCR) assay was established for semiquantitative detection of EBV sequences. The method was based on a nested PCR, using primers of the virus capsid antigen p23 region and an endpoint dilution. This method was carried out on 68 plasma samples, 68 samples of peripheral blood mononuclear cells and 5 cerebrospinal fluid samples of 39 patients with various diseases to evaluate the EBV‐genome copy number. Samples from patients suffering from infectious mononucleosis served as positive controls for active EBV infection. In 5 patients with infectious mononucleosis, high copy numbers of EBV genomes in peripheral blood mononuclear cells were detected within a range of 1,000–40,000 copies in 105 peripheral blood mononuclear cells. In contrast, samples from 19 latently infected persons either showed low copy numbers (10–100 in 105 peripheral blood mononuclear cells) or were EBV PCR negative. Comparable results were observed in seven renal transplant patients without any symptoms. The practical value of the semiquantitative detection of EBV DNA was demonstrated in three bone marrow transplant recipients. Two developed a lymphoproliferative disease associated with extremely high amounts of EBV DNA in plasma (16,000 and 50,000 copies/ml, respectively) and peripheral blood mononuclear cells (100,000 and 6.5 million copies in 105 peripheral blood mononuclear cells, respectively). The high EBV load in plasma and peripheral blood mononuclear cells was reduced dramatically after successful antiviral therapy in one case. The third bone marrow transplant recipient developed an EBV‐induced transverse myelitis with an increased number of EBV‐genome copies in peripheral blood mononuclear cells and EBV‐positive cerebrospinal fluid samples. After combined antiviral and immune therapy, the EBV‐genome copy numbers decreased and the patient recovered completely. These data demonstrate a good correlation between semiquantitative detection of EBV genomes and clinical findings. The method is recommended for the diagnosis of EBV‐associated diseases in patients after transplantation, as well as for monitoring the response to therapy. J. Med. Virol. 65:348–357, 2001. © 2001 Wiley‐Liss, Inc.
感染人类免疫缺陷病毒的患者发生与 Epstein-Barr 病毒相关的非霍奇金淋巴瘤。
影响因子：
20.3
作者：
Shibata,D;Weiss,LM;Hernandez,AM;Nathwani,BN;Bernstein,L;Levine,AM
通讯作者：
Levine,AM