Hill Charles E, Harris Shealynn B, Culler Elizabeth E, Zimring James C, Nolte Frederick S, Caliendo Angela M
Department of Pathology and Laboratory Medicine, Emory University School of Medicine, 1364 Clifton Road, Atlanta, GA 30322, USA.
Am J Clin Pathol. 2006 May;125(5):665-71. doi: 10.1309/ABEY-V2VK-E6DH-XAAA.
We compared the performance of a laboratory-developed 5'-nuclease real-time polymerase chain reaction assay and a commercial assay (LightCycler, Roche Diagnostics, Indianapolis, IN) for quantification of Epstein-Barr virus (EBV) DNA. Using standards provided by the manufacturer, the LightCycler assay was linear from 100 to 1 million copies per reaction. Based on dilution of a plasmid containing the amplicon, the laboratory-developed assay was linear from 22 to 45 million copies per reaction. Both assays detected 0.5 copies of genomic EBV DNA per reaction; both showed good reproducibility with coefficients of variation from 0.3% to 2.4% for the LightCycler and 1.8% to 5.1% for the laboratory-developed assay. For 31 whole blood specimens with measurable values in both assays, the viral load values obtained with the LightCycler averaged 2.3-fold higher than those obtained with the laboratory-developed assay. Testing 30 matched whole blood and plasma samples in the laboratory-developed assay showed whole blood viral load values averaged 10-fold higher than those for plasma. The LightCycler and laboratory-developed assays are sensitive and reproducible with broad linear ranges. Further clinical evaluation is needed to determine the viral load cutoff that is predictive of posttransplantation lymphoproliferative disorders.
我们比较了实验室研发的5'-核酸酶实时聚合酶链反应检测法与一种商业检测法（LightCycler，罗氏诊断公司，印第安纳波利斯，印第安纳州）对爱泼斯坦-巴尔病毒（EBV）DNA进行定量分析的性能。使用制造商提供的标准品，LightCycler检测法在每个反应100至100万拷贝数范围内呈线性。基于含有扩增子的质粒的稀释，实验室研发的检测法在每个反应2200万至4500万拷贝数范围内呈线性。两种检测法均能检测到每个反应0.5拷贝的EBV基因组DNA；两者均显示出良好的重复性，LightCycler检测法的变异系数为0.3%至2.4%，实验室研发的检测法的变异系数为1.8%至5.1%。对于31份在两种检测法中均有可测量值的全血标本，LightCycler检测法获得的病毒载量值平均比实验室研发的检测法高2.3倍。在实验室研发的检测法中对30对匹配的全血和血浆样本进行检测，结果显示全血病毒载量值平均比血浆高10倍。LightCycler检测法和实验室研发的检测法灵敏且可重复，线性范围广。需要进一步的临床评估来确定预测移植后淋巴细胞增生性疾病的病毒载量临界值。
