荧光定量PCR检测传染性单核细胞增多症不同血样本EBV的研究

目的]寻找荧光定量PCR检测传染性单核细胞增多症（IM）EBV DNA拷贝量的最佳样本。方法]采用荧光定量PCR分别检测了IM病人的外周血单个核细胞、全血基因组DNA及血浆的EBV DNA拷贝量，并比较各种方法的阳性率及拷贝量。结果]三种不同来源的样品荧光定量PCR方法的阳性率分别为76％、69％、23％。全基因组及单个核细胞组阳性率比较经x^2检验，差异无显著性，且两组拷贝量比较差异亦无显著性。血浆组阳性率及拷贝量与前两种方法比较差异均有显著性。结论]外周血单个核细胞是荧光定量PCR检测IM的EBV DNA量的最佳样本。

Assessment of Diagnostic Value with Real-time PCR for Measuring Epstein-Barr Virus Load in Whole Blood,Peripheral Mononuclear Cells and Plasm

Objective]Epstein-Barr virus （EBV） DNA load monitoring in blood has been shown to be essential for the diagnosis of infectious mononucleosis. The aim of this study was to evaluate the relative diagnostic value of EBV DNA load monitoring in whole blood, peripheral blood mononuclear cells （PBMCs） and plasma by LightCycler（r） real-time quantitative PCR （LC-PCR）. Methods]First, 52 patients who suffered from infectious mononucleosis were diagnosed . Second, EBV DNA load was determined prospectively by LightCycler（r） real-time quantitative PCR （LC-PCR） in the whole blood, PBMCs and plasma . Results] 76% .69% .23% samples were positive in PBMCs ,whole blood and plasma resepectively. EBV DNA load was 3.98±0. 64 log10copies/mL, 3.79±0.61 log10copies/mL ,2.71± 0.91 log10copies/mL resepectively . The frequency of positive EBV and the load was significantly different between PBMCs or whole blood and plasma P 〈0.05 ,but there was not significantly different between PBMCs and whole blood. P 〉0.05. Conclusion] The quantitative detection of EBV DNA load in PBMCs and whole blood appeared more sensitive than in plasma for infectious mononucleosis. Because EBV DNA extraction was time-consumed, so PBMCs was the most feasible blood compartments for the early diagnosis of infectious mononucleosis.