EB病毒VCA-IgA抗体及病毒DNA载量在系统性红斑狼疮患者中的表达

目的 检测EB病毒VCA-IgA抗体及病毒DNA载量在系统性红斑狼疮（SLE）患者外周血中的表达，探讨EB病毒与SLE的相关性。方法 分别采用酶联免疫吸附试验（ELISA）法和实时定量PCR方法检测EB病毒VCA-IgA抗体和EB病毒DNA载量。同时分析EB病毒VCA-IgA抗体与SLE患者的实验室指标抗Sm抗体、抗dsDNA抗体、补体C3和C4的相关性。结果 248例SLE患者中，49例为EBV-VCA-IgA抗体阳性，167例健康对照者中12例阳性，SLE患者阳性率明显高于对照组(19.8% vs 7.2%; P < 0.001)；SLE患者EB病毒载量也明显高于对照组。EB病毒VCA-IgA抗体阳性与抗Sm抗体、抗dsDNA抗体、补体C3和C4不相关。结论EB病毒感染与SLE相关，EB病毒VCA-IgA抗体阳性者有较高的DNA载量，SLE的发病危险性亦高，EB病毒重新活化与SLE活动有关。     

3项指标联合检测对儿童EB病毒感染的诊断价值

目的评价抗EB病毒衣壳抗原IgM抗体(VCA-IgM)、EB病毒衣壳抗原IgG抗体亲合力指数(RAI)、外周血异型淋巴细胞单项及联合检测对儿童EB病毒现症感染的诊断价值。方法收集2016年10月至2018年3月该院儿科收治的,采用酶联免疫吸附试验(ELISA)测定EB病毒VCA-IgM为阳性的209例患者作为研究对象。采用实时荧光定量聚合酶链反应检测EB病毒DNA,ELISA测定RAI,人工计数外周血异型淋巴细胞百分比。将标本分成EB病毒DNA阳性组和EB病毒DNA阴性组,采用SPSS19.0统计软件对各项指标进行非参数检验,通过Logistic回归和ROC曲线比较独立检测和联合检测各项指标在EB病毒感染诊断中的灵敏度和特异度。结果 EB病毒DNA阳性组的VCA-IgM、外周血异型淋巴细胞百分比均高于EB病毒DNA阴性组,RAI低于EB病毒DNA阴性组,差异有统计学意义(P0.05)。VCA-IgM、RAI、外周血异型淋巴细胞百分比三者均与EB病毒DNA相关(P0.05)。VCA-IgM、RAI、外周血异型淋巴细胞百分比三者单项检测的曲线下面积(AUC)分别为0.633、0.844、0.870,三者联合检测的AUC为0.875,高于各指标单项检测的AUC。结论诊断EB病毒感染的相关指标中,外周血异型淋巴细胞百分比最具有诊断学意义,优于VCAIgM、RAI,三者联合检测优于单项检测,有助于EB病毒感染及其他合并感染的诊断和防治。     

EB病毒感染与传染性单核细胞增多症的相关性分析

随着科技的不断进步,几乎所有医院都已配备自动化程度不同的血细胞分析仪,自动化程度越高的仪器分析参数越多,人工分片已严重缺乏,异型淋巴细胞检出率明显下降.过去传染性单核细胞增多症主要依据EB病毒抗体及血液中异型淋巴细胞数量进行诊断.近年来已有实验表明EB病毒DNA定量检测较抗体检测阳性率更高[1].本文主要探讨外周血中EB病毒DNA浓度与传染性单核细胞增多症发生率的相关性.     

儿童外周血EB病毒DNA检测的临床意义

目的探讨Epstein-Barr病毒(EB病毒)DNA检测的临床应用价值。方法应用荧光定量聚合酶链反应病毒核酸扩增方法,定量检测儿科以发热为主诉的患儿外周血EB病毒DNA载量。阳性病例进行临床分析和抗病毒治疗观察。结果检测522例间断和/或持续发热患儿,阳性(病毒载量≥50拷贝/ml)102例,阳性率19.5%。病毒载量最高4.44×106拷贝/ml,最低4.3×102拷贝/ml。阳性病例疾病病种较多。部分阳性病例抗病毒治疗显效。结论静脉血EB病毒DNA检测是可行的实验室诊断方法;儿科以发热为主诉的患儿可能有19.5%左右存在EB病毒感染或隐性感染。EB病毒DNA检测阳性患儿应适当抗病毒治疗并随访。     

乙型肝炎病毒前S1抗原与乙肝DNA定量联合检测的临床应用

目的 检测乙型肝炎不同血清学标志物模式患者HBVDNA含量,并与前S1抗原进行比较分析.方法 运用酶联免疫吸附测定法、荧光定量聚合酶链反应(PCR)技术对286例各型乙型肝炎患者进行HBV-DNA含量及血清学标志物检测结果进行相关性分析.结果 血清学检测乙肝表面抗原(HBsAg)+乙肝病毒e抗原(HBeAg)+乙肝病毒核心抗体(抗HBc)阳性(大三阳)患者HBV-DNA阳性率明显高于其他类型(P＜0.01),且其病毒载量显著高于其他组HBsAg+乙肝病毒e抗体(HBeAb)+抗HBc阳性(小三阳)阳性检出率也较高,其病毒载量高于其他组.结论 PreS1抗原与乙肝DNA定量联合检测具有重要的临床应用价值.     

EB病毒6项抗体联合检测在临床诊断中的应用

目的探讨酶联免疫吸附试验(ELISA)检测EB病毒抗体谱的临床应用价值。方法采用ELISA法对2 526例该院住院患者进行EB病毒6项抗体联合检测,结合病理诊断结果对EB病毒6项抗体联合检测结果进行统计分析,探讨EB病毒感染与疾病的关系。结果 2 526例住院患者中有32例经病理诊断为鼻咽癌,占1.27%。抗EB病毒衣壳抗原抗体IgA(EBVCAIgA)、抗EB病毒早期抗原抗体IgG(EBVEA-IgG)、抗EB病毒早期抗原抗体IgA(EBVEA-IgA)阳性共18例,阳性率为56.25%;EBVCA-IgA、EBVEA-IgG阳性10例,阳性率为31.25%;EBVCA-IgA、EBVEA-IgA阳性3例,阳性率为9.38%。2 526例住院患者中有52例为儿童患者,上述3种抗体阳性模式所涉及的疾病主要为呼吸、消化、血液等系统疾病,且以呼吸系统疾病为主(57.7%),但均未诊断为鼻咽癌。结论 EB病毒抗体阳性模式中EBVCA-IgA、EBVEA-IgG、EBVEA-IgA阳性或EBVCA-IgA、EBVEA-IgG阳性或EBVCA-IgA、EBVEA-IgA阳性在成年人中可诊断为鼻咽癌,而在儿童中出现以上检测结果,则不一定能诊断为鼻咽癌。但EB病毒6项抗体联合检测对儿童EB病毒感染引起的相关疾病具有参考价值。     

乙型肝炎病毒血清学标志物与乙肝DNA相关性探讨

目的探讨乙肝病毒DNA（HBV—DNA）病毒载量与各型乙型肝炎血清标志物之间的关系。方法运用酶联免疫吸附测定法、荧光定量聚合酶链反应（PCR）技术对350例各型乙型肝炎患者进行HBV—DNA含量及血清学标志物检测（两对半）。结果血清学检测乙肝表面抗原（HBsAg）＋乙肝病毒e抗原（HBeAg）＋乙肝病毒核心抗体（抗HBc）阳性（大三阳）患者HBV—DNA阳性率明显高于其他类型（阳性率95．7％），且其病毒载量显著高于其他组。HBsAg＋乙肝病毒e抗体（HBeAb）＋抗HBc阳性（小三阳）阳性检出率也较高（阳性率54．0％）其病毒载量高于其他组。结论HBV—DNA与HBeAg的存在明显正相关。在临床工作中，不仅要应用乙肝血清标志物来判断HBV是否在体内复制，更要结合PCR检测技术来测定HBV—DNA含量。     

67例成人EB病毒感染临床及实验室特征分析

目的:检测成人Epstein-Barr病毒(EB病毒)感染患者的EB病毒相关抗体及EB病毒DNA、血常规、血生化指标,对其临床症状进行分析,探讨成人EB病毒感染的临床特点。方法:收集67例成人EB病毒感染患者的临床资料及实验室检测结果,并将其按年龄分为青中年组(18～59岁,19例)和老年组(≥60岁,48例);另收集同期的49名健康体检者,按前述年龄分为2组,分别作为年龄对照组。分析成人EB病毒感染的临床特点及不同年龄段患者间的异同。结果:67例成人EB病毒感染患者中,最常见的症状为发热(37例,55.2%);其次为淋巴结肿大(13例,19.4%)、脾肿大(9例,13.4%)、肝脏肿大(8例,11.9%)、咽部充血(6例,9.0%)和皮疹(5例,7.5%)。EB病毒感染的青中年患者与老年患者相比较,发热(84.2%比43.8%)、咽部充血(26.3%比2.1%)、皮疹(21.1%比2.1%)、肝脏肿大(31.6%比4.2%)的临床症状明显(χ2值分别为9.012、7.057、4.612、7.295,P均0.05)。19例青中年患者中有2例伴发恶性肿瘤性疾病;而48例老年患者中有2例(4.1%)伴发嗜血细胞淋巴组织细胞增生症,17例(34.7%)伴发恶性肿瘤性疾病,2组间的恶性肿瘤患病构成比差异有统计学意义(χ2=4.15,P=0.042)。EB病毒感染老年组的T淋巴细胞总数、CD4+T淋巴细胞计数、B淋巴细胞计数均低于健康老年组(t分别为4.525,5.960,2.999,P均0.05),而EB病毒感染青中年组的淋巴细胞亚群数与健康青中年组比较,差异无统计学意义(P0.05)。青中年EB病毒感染患者与老年患者比较,EB病毒相关抗体表达水平及肝功能指标差异无统计学意义(P均0.05)。结论:成人EB病毒感染患者的临床表现各异,青中年患者的急性感染症状较老年患者明显,而老年患者的细胞免疫及体液免疫功能降低,且EB病毒相关肿瘤患病者构成比高。     

EB病毒感染的核酸检测:标本类型的选择

近年来的研究表明,EB病毒(epstein-barr virus,EBV)感染与多种疾病的发生有密切的关系.由于EB病毒独特的感染方式,其实验室诊断特别是核酸检测显得尤为重要.本文就EB病毒的生物学特性、感染方式、抗体产生,特别是核酸检测中标本类型的选择、标准化以及未来的检测方向等问题进行了述评和展望,以期为EB病毒感染相关疾病的诊断和预后评判提供更准确的核酸检测结果.     

EB病毒近期感染与系统性红斑狼疮的关联性研究

目的：检测系统性红斑狼疮（SLE）患者体内 EB 病毒衣壳抗体（EBV-VCA-IgG、IgA）滴度及病毒DNA载量,探讨SLE患者EB病毒感染情况及病毒再发感染后的激活对SLE总体疾病活动度的影响。方法收集80例SLE患者、118例非SLE的风湿患者、33例正常对照组血标本,运用酶联免疫吸附实验（ELISA）测EB病毒抗体,实时荧光定量PCR测EBV-DNA载量。运用卡方检验比较各组病毒抗体或DNA阳性率差别,非参数检验比较各组病毒抗体滴度、红细胞沉降率、SLE疾病活动度（SLEDAI）等,Spearman相关分析病毒抗体滴度或DNA载量与SLEDAI的关联性。结果（1）抗体阳性率：SLE组EBV-IgA阳性率显著高于非SLE组及正常对照组（57.5%vs.16.96%,χ2＝37.054,P＜0.000;57.5%vs.15.15%,χ2＝16.92,P＜0.000）,并与SLE的发病有显著的相关性（r＝7.57,P＜0.05）,但IgG抗体阳性率与对照组相比无差别。抗体滴度：SLE组EBV-IgA及IgG滴度显著高于正常对照组[EBV-IgA：1.11（2.268） vs.0.495（0.856）,Z＝-2.836,P＝0.005;EBV-IgG：5.391（6.038）vs.4.121（5.231）,Z＝-2.49,P＝0.013];（2） SLE组EBV-IgA及IgG滴度与对应的SLEDAI未发现明显的相关性（EBV-IgA：r＝0.164,P＝0.195;EBV-IgG：r＝-0.09,P＝0.576）。也未发现EBV-IgA阳性患者在皮疹、关节炎、肝损害、SLEDAI、低补体血症等方面与阴性患者的差别,但红细胞沉降率却明显高于阴性患者[47（82.75）mm/1 h vs.26.5（46.25）mm/1 h, Z＝-3.028,P＝0.002];（3）80例SLE患者中有42例随访了IgA表型变化,但未发现表型变化对SLEDAI未见明显差异的影响。（4）SLE患者EBV-DNA阳性率也显著高于非SLE风湿病组（27.5% vs.12.8%,χ2＝6.712,P＜0.05）,SLE患者EB病毒DNA载量异常与正常者的SLEDAI比较无统计学意义[分别为15（22） vs.13（20.25）,Z＝-0.573,P＝0.567],并且异常DNA载量值与对应的SLEDAI之间也未见明显的相关性。结论 SLE患者EBV抗体及DNA的高表达,提示EB病毒与SLE密切相关,但是近期感染或再发感染的激活（EBV-IgA或DNA）未影响到SLE总体的疾病活动度变化。     

Limitations of polymerase chain reaction testing for diagnosing acute Epstein-Barr virus infections

Clinicians use molecular tests to detect Herpesviridae from blood without fully appreciating limitations of testing. Studies are needed to enhance our understanding of the impact of Herpesviridae latency on molecular testing. We retrospectively performed quantitative Epstein-Barr virus (EBV) on sera from patients between the ages of 1 and 30 who demonstrated serologic evidence of acute EBV (n = 50) or remote EBV (n = 50) infection. Epstein-Barr virus DNA was detected in 70% of acutely infected and 4% of remotely infected patients. Sera from acutely infected patients had higher EBV copy number than convalescent sera. Our results suggest that serology should be performed as the initial diagnostic test for acute EBV. The role for polymerase chain reaction in immunocompromised patients with impaired antibody responses or as a 2nd-line diagnostic test when serologic results are equivocal deserves further study.     

Identical IgM antibodies recognizing a glycine-alanine epitope are induced during acute infection with Epstein-Barr virus and cytomegalovirus.

We studied antibody production in serial serum samples from patients with acute Epstein-Barr virus (EBV) and cytomegalovirus (CMV) infections. Sera were analyzed both by enzyme-linked immunosorbent assay (ELISA) using a synthetic peptide (P62) derived from the glycine-alanine repeating region of the Epstein-Barr nuclear antigen (EBNA-1) and by immunoblotting. In prior studies, we have shown that patients with acute EBV infection make IgM antibodies that react with this peptide, that recognize a viral-specific protein (EBNA-1), and that bind with a number of proteins present in uninfected cells; however, antibody binding to these autoantigens was inhibited by the peptide. IgG antibodies reactive with the peptide did not appear until months after the disease and were specific for the EBNA-1 protein. We now find that patients with acute CMV infection but not those with acute infections from a variety of other nonherpes organisms also produce IgM antibodies that recognize the EBV-derived peptide P62. These antibodies also appear to recognize the same cellular proteins as the EBV-induced IgM antibodies. The IgM antibodies appeared in all acutely infected CMV patients studied and occurred both in patients with previous EBV infections and in one patient studied who had not previously been exposed to EBV. It appears that infection with EBV or CMV can induce the synthesis of a very similar or identical set of IgM antibodies.     

EB病毒与其相关淋巴瘤的研究进展

EB病毒（Epstein-Barr virus,EBV）属于r-DNA疱疹病毒科,是最早被发现与人类肿瘤相关的病毒。大量研究发现,EBV的感染与多种淋巴瘤的发生有关,如霍奇金淋巴瘤、伯基特淋巴瘤、NK/T细胞淋巴瘤、HIV相关的淋巴瘤和弥漫大B细胞淋巴瘤等。近年来研究表明,EBV潜伏感染的基因表达产物对淋巴瘤的发生发展起到促进作用,并为EBV相关淋巴瘤的治疗提供了依据。现就EBV在几种常见淋巴瘤中的作用机制及治疗的研究进展做如下综述。     

淋巴组织增生性疾病患者外周血EBV DNA定量检测临床意义的研究进展

目前发现爱泼斯坦巴尔病毒(Epstein-Barr virus,EBV)与多种淋巴组织增生性疾病的关系越来越密切,主要包括EBV相关淋巴瘤、EBV阳性淋巴组织增殖性疾病(EBV+LPD)以及传染性单核细胞增多症(infectious mononucleosis,IM)等,以往研究认为EBV+LPD及IM的外周血EBV ...     

Clinically severe Epstein-Barr virus encephalitis with mild cerebrospinal fluid abnormalities in an immunocompetent adolescent: a case report

A 15-year-old boy developed Epstein-Barr virus (EBV) encephalitis, a rare complication of infectious mononucleosis. The severe clinical picture and the marked neuroimaging changes were in contrast with mild cerebrospinal fluid abnormalities: leukocyte count was normal and protein level was only slightly elevated. EBV DNA was detected in cerebrospinal fluid by polymerase chain reaction.     

Functionally active Epstein-Barr virus-transformed follicular dendritic cell-like cell lines

Follicular dendritic cells (FDC) are unique nonlymphoid cells found only in germinal centers. FDC can be distinguished from other accessory cells based on a characteristic set of cell surface markers. It is known that FDC are able to rescue germinal center B cells from apoptosis. To investigate the role of FDC in the process of selection and maturation of B cells during germinal center reactions, we tried to establish factor-independent immortalized FDC-like cell lines. Because freshly isolated FDC express the Epstein-Barr Virus (EBV) receptor CD21, we attempted EBV transformation on isolated FDC. After incubation of FDC-enriched cell populations with EBV, cell lines were obtained consisting of slowly duplicating very large cells. These cell lines have a fibroblast-like morphology but could be clearly distinguished from several human fibroblast cell lines by displaying a different phenotype including intercellular adhesion molecule 1, CD40, and CD75 expression. Detection of the EBV-encoded proteins latent membrane protein 1 and Epstein-Barr virus nuclear antigen 2 in our FDC-like cell lines implicated successful EBV transformation. FDC-like cells are able to bind nonautologous B cells and preserve the latter from apoptosis. The binding of B cells to FDC-like cells is dependent on adhesion via lymphocyte function-associated antigen 1/intercellular adhesion molecule 1 and closely resembles the pattern of emperipolesis as described by others. These data demonstrate that FDC can be successfully infected by EBV, and that the cell lines obtained share phenotypic and functional characteristics with freshly isolated FDC.     

Epstein-Barr virus susceptibility of normal human B lymphocyte populations

Human blood and tonsil B lymphocytes were fractionated on density gradients and tested for virus binding and penetration into the cells. Epstein-Barr Virus (EBV) transformation was detected by immunofluorescence staining for EBV-determined nuclear antigen (EBNA). EBV bound to and penetrated all B cell populations, but only the high density populations were transformed. Activated B lymphocytes were found in the low density fractions and these cells were resistant to EBV infection. Infected and noninfected B lymphocytes were density-analyzed during in vitro culture. A spontaneous, not virus-induced, density decrease was found to precede the production of EBNA. Cells remaining at high density never expressed EBNA. The results suggest that EBV can transform only small resting B lymphocytes and that a virus-independent activation of the infected cells induces the EBNA production and transformation.     

Hemophagocytic syndrome due to Epstein-Barr virus and cytomegalovirus coinfection in a patient on adalimumab

IntroductionHemophagocytic syndrome (HPS) is a rare but potentially fatal complication of viral infections. Epstein-Barr virus (EBV) and cytomegalovirus (CMV) often infect patients receiving TNF-alpha inhibitors (TNF-α inhibitors). While EBV and CMV are well established infections for the development of infectious mononucleosis, coinfection with EBV and CMV is common among immunosuppressed patients and can result in a fatal course. In addition, such viral infections can cause HPS. To the best of our knowledge, we present here the first report of HPS induced by EBV and CMV coinfection during anti-TNFα inhibitor use.Case reportA 23-year-old man hospitalized with fever, elevated liver enzymes, lymphadenopathy, and hepatosplenomegaly was diagnosed with HPS associated with EBV and CMV coinfection while using adalimumab. No clinical improvement was observed after discontinuation of adalimumab. HPS complicated by EBV and CMV coinfection was finally diagnosed, and immediate administration of ganciclovir and prednisone was considered to have prevented a lethal clinical outcome.ConclusionFor cases showing unexplained fever, elevated liver enzymes, and lymphadenopathy while using anti-TNFα inhibitors, screening for EBV and CMV coinfection should be encouraged. In addition, HPS should be considered in patients with EBV and/or CMV infection receiving anti-TNFα inhibitors to facilitate early definitive therapy.     

Comparative evaluation of the new ARCHITECT EBV assays considering different testing algorithms

In the current evaluation, Epstein-Barr virus (EBV) serology was performed on 1113 routine serum samples. Although the initial request for all samples from the general practitioner was EBV IgM testing, 80.9% were classified as past infections. The ARCHITECT^® viral capsid antigen (VCA) IgM, VCA IgG, and EBV nuclear antigen (EBNA) 1 IgG assays showed good results for sensitivity and specificity, being 100.0%, 98.3%, and 100.0% and 99.9%, 95.4%, and 99.6%, respectively. Using an algorithm based on initial EBNA-1 IgG testing, followed by VCA IgG and IgM for samples that were not EBNA-1 IgG reactive, the number of tests per sample could be reduced to nearly 50% compared to parallel testing. The high sensitivity and specificity of the ARCHITECT^® EBNA-1 IgG assay in combination with a low number of grayzone results are a precondition for the chosen test algorithm. Thus, the newly developed ARCHITECT^® EBV panel is suitable for accurate and cost-efficient EBV serology in a routine clinical laboratory.     

Epstein-Barr virus quantitation by real-time PCR targeting multiple gene segments: a novel approach to screen for the virus in paraffin-embedded tissue and plasma

Epstein-Barr Virus (EBV) infects nearly all humans and then persists for the life of the host. In some people who later develop cancer, EBV DNA is present within malignant cells and circulates at elevated levels in the plasma. In the current study, we validated five novel quantitative polymerase chain reaction (Q-PCR) assays targeting disparate but highly conserved segments of the EBV genome (BamH1W, EBNA1, LMP1, LMP2, and BZLF1). Each assay was sensitive to as few as 50 copies of EBV DNA per reaction and was linear across at least four orders of magnitude. When applied to paraffin-embedded tissues in concert with EBV-encoded RNA (EBER) in situ hybridization, the BamH1W and EBNA1 assays were the most informative, while use of the entire battery of EBV PCR assays may help identify genomic polymorphisms or deletions. Higher viral loads were found in the 17 EBER-positive compared with the 13 EBER-negative tumors (means 84,978 versus 22 copies of EBV per 100,000 cells, respectively). The five Q-PCR assays were also informative in plasma samples where EBV was measurable in all nine patients with lymphoma or infectious mononucleosis, whereas EBV was undetectable in all nine healthy controls. The findings suggest that Q-PCR is an effective method of distinguishing disease-associated virus from incidental virus in paraffin-embedded tissue and in plasma samples.