外周血单个核细胞中和血浆中EB病毒核酸载量的对比分析

Comparative analysis of Epstein-Barr virus nucleic acid loads in peripheral blood mononuclear cells and plasma

目的探讨EBV DNA检测中标本的合理选择。方法收集2017年1月至6月在北京友谊医院确诊为EB病毒感染的患者，共117例，其中传染性单核粒细胞增多症（IM）44例，EB病毒相关噬血细胞综合征（HLH）36例，移植后淋巴细胞增殖性异常（PTLD）37例，年龄在6个月至28岁。采用实时荧光定量PCR法，定量检测外周血PBMC中和血浆中EBV DNA载量（中位数，四分位数表示），不同标本类型间病毒载量比较采用非参数秩和检验（Mann－Whitney检验），相关性分析采用Spearman相关性分析。结果IM和PTLD患者PBMC中EBV DNA载量分别为53 600（7 875，626 500）拷贝/ml和114 000（3 396，590 500）拷贝/ml，显著高于血浆中的载量4 500（675，8 600）拷贝/ml和0（0，0）拷贝/ml，M－W值分别为372.5和30.5，P均〈0.001，差异有统计学意义，而HLH患者PBMC和血浆中EBV DNA载量分别为5 100（1 425，170 000）拷贝/ml和13 500（1 303，152 500）拷贝/ml，M－W值为646.5，P＝0.991，差异无统计学差异。Spearman相关性分析显示IM和HLH患者中PBMC和血浆EBV DNA载量有较好的相关性，r值分别为0.548和0.400，P均〈0.05，而PTLD患者中PBMC和血浆EBV DNA载量无相关性，r值为0.308，P〉0.05。结论对于EBV感染的诊断与监测，不同疾病推荐的标本类型有所差异，IM和HLH中EBV DNA定量检测推荐采用血浆或血清标本，而PTLD则推荐同时检测PBMC和血浆标本，临床上应根据疾病的种类合理选择标本类型。

ObjectiveTo find the rational selection of specimens for the detection of Epstein-Barr virus （EBV） DNA.MethodsA total of 117 patients were diagnosed with EBV infection at Beijing Friendship Hospital from January to June 2017, including 44 patients with infectious mononucleosis （IM）, 36 patients with EBV-associated hemophagocyticlymphohistiocytosis （HLH） and 37 patients with post-transplant lymphoproliferative disorders （PTLD）. Patients were aged from 6 months to 28 years. EBV DNA loads （median and quartile） in peripheral blood mononuclear cells （PBMC） and plasmawere detected by real-time quantitative PCR. The viral loads of different specimen types were compared by nonparametric rank sum test （Mann-Whitney test, M-W test）. Spearman correlation analysis was performed for correlation analysis.ResultsTheEBV DNA loads in PBMC of IM and PTLD were 53 600 （7 875, 626 500） copies/ml and 114 000 （3 396, 590 500） copies/ml, which were significantly higher than those in plasma[4 500 （675, 8 600）copies/ml and 0（0, 0）copies/ml, respectively]. The M-W values were 372.5 and 30.5 respectively （both P〈0.001）, which indicated statistically significant differences. However, the EBV DNA loads in PBMC and plasma of HLH were 5 100 （1 425, 170 000） copies/ml and 13 500 （1 303, 152 500） copies/ml. The M-W value was 646.5 （P=0.991）, which indicated no statistically significant difference. Spearman correlation analysis showed good correlations of EBV DNA loads between PBMC and plasmain IM and HLH, and the r values were 0.548 and 0.400, respectively （both P〈0.05）, while the correlation of EBV DNA loads between PBMC and plasma in PTLD was poor, and the r value was 0.308 （P〉0.05）.ConclusionsFor the diagnosis and monitoring of EBV infection, the types of specimens recommended by different diseases are different. Plasma or serum specimens are recommended for quantitative detection of EBV DNA in IM and HLH, while PBMC and plasma specimens are recommended in PTLD.Clinically, the type of specimen should be chosen reasonably according to the type of disease.（Chin J Lab Med, 2018, 41： 755-758）