EB病毒核酸及血清学检测在EB病毒相关疾病中的应用研究

摘要

目的 通过对同一对象血标本进行EB病毒核酸及EB病毒血清学的检测,评估两者之间的关联及在不同疾病中的应用价值.方法 通过回顾2014年5月至2015年11月在华中科技大学同济医学院附属同济医院住院的397例EB病毒感染的病例并收集其住院期间外周血样本,另外收集120名健康体检者的EDTA抗凝外周血标本,用实时荧光定量PCR检测外周血单个核细胞(PBMC)及血浆中的EB病毒核酸,用酶联免疫吸附试验检测血浆EB病毒抗体5项(抗衣壳抗原IgA抗体VCA IgA、抗衣壳抗原IgM 抗体VCA IgM、抗衣壳抗原IgG抗体VCA IgG、抗早期抗原IgG抗体(弥散型)EA(D)IgG、抗核心抗原IgG抗体EBNA IgG),使用Kappa一致性检验及Spearman相关性分析评估两类检测结果之间的一致性及相关性,并按诊断进行分组,比较不同疾病中各检测项目的阳性率.结果 血浆VCA IgG在疾病组及健康对照组中的阳性率分别为94.2% (374/397)及93.3% (112/120) (χ2=0.125,P=0.67);EBNA IgG在两组中的阳性率分别为95.4% (379/397)及95.0% (114/120)(χ2=0.045,P=0.807);而VCA IgM在两组中的阳性率分别为5.5% (22/397)及0% (0/120) (χ2=6.9,P<0.01);VCA IgA在两组中的阳性率分别为43.3% (172/397)及9.2% (10/120) (χ2=49.5,P<0.01);EA(D) IgG在两组中的阳性率分别为42.0% (167/397)及7.5% (9/120) (χ2=49,P<0.01);PBMC中EB病毒核酸在两组中的阳性率分别为65.5% (260/397)及16.7% (20/120) (χ2=88.5,P<0.01);血浆中EB病毒核酸在两组中的阳性率分别为45.8% (182/397)及5.0% (6/120) (χ2=66.4,P<0.01).相同标本单个核细胞EB病毒核酸与血浆VCA IgM、VCA IgA、EA(D)IgG检测结果的一致性较差(Kappa值分别为0.073、0.147、0.073);血浆EB病毒核酸与血浆VCA IgM、VCA IgA、EA(D) IgG检测结果的一致性同样较差(Kappa值分别为0.144、0.369、0.288).进一步分析发现,在结外NK/T细胞淋巴瘤、EB病毒相关嗜血淋巴组织增生症、慢性活动性EB病毒感染及传染性单核细胞增多症分组中,EB病毒核酸阳性率高于90%;而在鼻咽癌患者中,VCA IgA及EA(D) IgG抗体阳性率高于EB病毒核酸.结论 EB病毒核酸与EB病毒血清学检测结果之间的一致性较差,EB病毒核酸与EB病毒血清学在鼻咽癌、淋巴瘤、传染性单核细胞增多症等不同疾病中的临床应用价值存在差异.%Objective To investigate the relationship between Epstein-Barr virus (EBV) DNA and EBV serology markers and evaluate the clinical application values in different diseases.Methods Plasma samples from 397 diagnosed EBV infection-associated patients and 120 health donors from May 2014 to November 2015 in Wuhan Tongji Hospital were collected.Real-time fluorescent quantitative PCR was performed to detect the levels of EBV-DNA in peripheral blood mononuclear cell and plasma.ELISA was used to detect VCA IgA,VCA IgM,VCA IgG,EA(D) IgG and EBNA IgG antibodies in plasma.The positive rate of EBV-DNA and EBV antibodies were counted in each group according to the detection threshold.Kappa statistic and Spearman sank correlation test were used to analysis the correlation and uniformity between EBV-DNA and EBV serology indicators.Results The positive rate of VCA IgG in patient and health control was 94.2% (374/397) and 93.3% (112/120) respectively (χ2=0.125,P=0.67);The positive rate of EBNA IgG in patient and health control was 95.4% (379/397) and 95.0% (114/120) respectively (χ2=0.045,P=0.807);but the positive rate of VCA IgM was 5.5% (22/397) and 0% (0/120) respectively (χ2=6.9,P<0.01);The positive rate of VCA IgA was 43.3% (172/397) and 9.2% (10/120) respectively (χ2=49.5,P<0.01);The positive rate of EA(D) IgG was 42.0% (167/397) and 7.5% (9/120) respectively (χ2=49,P<0.01).The positive rate of EBV-DNA was 65.5% (260/397) and 16.7% (20/120) respectively (χ2=88.5,P<0.01);The positive rate of EBV-DNA in plasma was 45.8% (182/397) and 5.0% (6/120) respectively (χ2=66.4,P<0.01).Furthermore,the uniformity and Spearman correlation analysis showed that there was no significant correlation between EBV-DNA and EBV serology indicators.The correlation analysis between PBMC EBV-DNA and VCA IgM,VCA IgA,EA(D) IgG showed the Kappa was 0.073,0.147,0.073,respectively;the correlation analysis between plasma EBV-DNA and VCA IgM,VCA IgA,EA(D) IgG showed the Kappa was 0.144,0.369,0.288,respectively.Thus,the patients were divided into different groups according to the discharge diagnosis,it was observed that the positive rates of EBV-DNA is more than 90% in extra-nodal NK/T cells lymphoma,EBV-associated hemophagocytic lymphoid tissue hyperplasia,chronic active EBV infection and infectious mononucleosis.In nasopharyngeal carcinoma patients,the positive rate of EBV antibodies (VCA IgA and EA(D) IgG) were higher than the detection of EBV-DNA.Conclusions There was no significant correlation between EBV-DNA and EBV serology markers for the same sample.The clinical application values of EBV DNA and EBV serology markers were not identical in nasopharyngeal carcinoma,extra-nodal NK/T cells lymphoma,infectious mononucleosis and EBV-associated hemophagocytic lymphoid tissue hyperplasia.