Development of a real-time quantitative assay for detection of Epstein-Barr virus
Development of a real-time quantitative assay for detection of Epstein-Barr virus
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DOI：
10.1128/jcm.38.2.712-715.2000
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发表时间：
2000-02-01
影响因子：
9.4
通讯作者：
Osterhaus, ADME
中科院分区：
文献类型：
--
作者：
Niesters, HGM;van Esser, J;Osterhaus, ADME
利用实时荧光PCR技术，建立了一种快速、灵敏、特异、重复性好的检测血浆中EB病毒DNA的方法。该方法允许我们使用基于吸收的两种样品制备方法在每毫升100至10（7）个DNA拷贝的范围内筛选血浆和血清样品。RI精密度研究得出两种方法的平均变异系数小于12%，标准曲线的回归系数最小为0.98。我们在19.2%的无EBV感染症状的免疫抑制实体器官移植患者的血浆样本中检测到EBV DNA，平均载量为440拷贝/ml，在所有诊断为移植后淋巴组织增生性疾病的移植患者中均可检测到EBV DNA，平均载量为544，570拷贝/ml，在非免疫抑制对照组的健康个体中未检测到EBV DNA，而在传染性单核细胞增多症患者中可检测到平均每毫升6100个拷贝。进一步的研究表明，肝素化血浆的抑制作用可以通过使用硅藻土作为吸收剂的提取方法有效地去除。
With the use of real-time PCR, we developed and evaluated a rapid, sensitive, specific, and reproducible method for the detection of Epstein-Barr virus (EBV) DNA in plasma samples. This method allowed us to screen plasma and serum samples over a range between 100 and 10(7) copies of DNA per mi using two sample preparation methods based on absorption. ri precision study yielded an average coefficient of variation for both methods of less than 12%, with a coefficient of regression for the standard curve of a minimum of 0.98. We detected EBV DNA in 19.2% of plasma samples from immunosuppressed solid-organ transplant patients without symptoms of EBV infections with a mean load of 440 copies per mi, EBV DNA could be detected in all transplant patients diagnosed with posttransplant lymphoproliferative disorder, with a mean load of 544,570 copies per mi, No EBV DNA could be detected in healthy individuals in nonimmunosuppressed control groups and a mean of 6, 100 copies per mi could be detected in patients with infectious mononucleosis. Further studies revealed that the inhibitory effect of heparinized plasma could be efficiently removed by use of an extraction method with Celite as the absorbent.