EBV-DNA检测在血液肿瘤性疾病中的应用价值

EB病毒(EBV)属人类疱疹病毒，与人类多种疾病相关联。目的 本研究探讨EB病毒感染与恶性血液病的关系。方法 应用PCR法检测38例血液肿瘤性疾病患儿骨髓细胞EBV-DNA。结果 38例患儿骨髓EBV-DNA阳性检出率61％(23／38)。5例恶性淋巴瘤患儿均为阳性，急性白血病(AL)、骨髓增生异常综合征(MDS)、再生障碍性贫血(AA)、特发性血小板减少性紫癜(ITP)患儿阳性率较高，其余病例均为阴性。对照组10例非血液病发热患儿仅1例阳性。结论 EBV感染与血液肿瘤性疾病的发生、发展及愈后关系密切。EBV感染可促使病情恶化或难治。同时对伴有不明原因发热的患儿除考虑肿瘤性发热外，应警惕因免疫功能异常导致的EBV易感因素，尽早动态检测EBV-DNA，及时合理治疗。由于病人免疫力低下，对EBV病毒的免疫应答功能降低，抗体反应法作EBV常造成假阳性结果。本方法简便、快速、特异、敏感，具有重要的临床应用价值。     

EBER原位杂交用于骨髓涂片中检测EB病毒

EB病毒(EBV)感染是小儿比较常见的病毒感染性疾病,感染时其症状轻重不一,可累及全身多个系统,引发多种疾病.目前公认EBV感染与三大类儿童血液系统疾病有关:淋巴系统疾病、造血系统疾病、自身免疫性疾病.我科与病理科联合开展了EBER(EBV编码的小RNA)原位杂交在骨髓涂片中检测EBV.     

EB病毒感染与淋巴增殖性疾病

EB病毒(EBV)是一种能诱发肿瘤的疱疹病毒.多数情况下,由于机体的免疫系统能严格限制病毒颗粒的增殖,受感染者终生携带病毒而不发病.少数情况下如机体免疫功能明显降低,则可引起严重的相关性疾病,特别是良性或恶性的淋巴增殖性疾病.近年来,有关EBV在人体内生存的机制、感染后的免疫调控、相关疾病的发病机制等方面均有了新的认识.现就近年来EBV与淋巴增殖性疾病的有关进展情况进行介绍.     

肿瘤

05 0 641　 EBV- DNA检测在血液肿瘤性疾病中的应用价值 /李　戈…∥中国小儿血液 .- 2 0 0 4 ,9( 2 ) .- 69～ 70结果提示 EBV感染与血液肿瘤性疾病的发生、发展及愈后关系密切。EBV感染可使其病情恶化或难治。同时对伴有不明原因发热的患儿除考虑肿瘤性发热外 ,应警惕因免疫功能异常导致的EBV易感因素 ,尽早动态检测 EBV- DNC,及时合理治疗。参 4(何秀兰 )0 5 0 642　儿童造血系统恶性肿瘤的免疫治疗 /吴　王月∥中国小儿血液 .- 2 0 0 4 ,9( 4 ) .- 1 91～封 30 5 0 643　多学科协作在儿童实体瘤诊断治疗整体方案中作用探讨 /汤…     

EB病毒感染与血液病

某些病毒感染可引起血液系统异常或导致血液系统疾病，如疱疹病毒中的EB病毒（EBV）、卡波西肉瘤病毒（KSHV或人类疱疹病毒-8）、巨细胞病毒（CMV）、细小病毒B19及人类免疫缺陷病毒（HIV）等。EBV、KSHV、CMV等疱疹病毒感染与宿主免疫状态密切相关。全球90％以上成人曾感染过EBV，一般均无临床症状，其体内少数B淋巴细胞中有潜伏EBV存在。     

EB病毒的研究进展

EB病毒 (Epstein Barrvirus ,EBV)是一个具有复杂基因组的人类疱疹病毒 ,全球 90 %以上的人受到过该病毒的感染。由于EBV对人类的普遍易感性和与多种人类疾病尤其是恶性肿瘤密切相关 ,EBV感染越来越受到人们的关注。研究EBV的免疫学特性、免疫系统对EBV感染的控制作用及EBV逃避机体免疫应答的机制、以及EBV感染相关疾病致病机理已成为近来重要的研究课题。一、EBV分子生物学特点与功能(一 )形态与结构1 96 4年EpsteinAchong和Barr[1] 首次从淋巴瘤细胞株分离出疱疹样病毒 ,命名为EB病毒。EBV属于γ疱疹病毒科 ,形态与其他人…     

儿童EB病毒原发性感染血浆病毒DNA拷贝数特征

目的探讨儿童EB病毒(EBV)原发性感染血浆病毒DNA拷贝数特征及其变化规律。方法回顾性病例总结。选择2017年9月1日至2018年9月30日于复旦大学附属儿科医院诊断为EBV原发性感染的571例患儿为研究对象, 收集患儿病毒实验室指标及临床信息等资料。根据血浆EBV DNA结果分为阳性组和阴性组, 根据EBV DNA载量将阳性组分为高载量组和低载量组。组间差异采用χ2检验、Wilcoxon秩和检验进行比较。结果 571例EBV原发性感染患儿中男334例、女237例, 初诊年龄3.8(2.2, 5.7)岁。阳性组255例(44.7%)、阴性组316例(55.3%)。阳性组发热、肝和(或)脾肿大、转氨酶升高的患儿比例均高于阴性组[235例(92.2%)比255例(80.7%)、169例(66.3%)比85例(26.9%)、144例(56.5%)比120例(38.0%), χ2=15.22、96.80、18.27, 均P<0.001];阳性组中有70例进行了随访, 随访时间46(27, 106)d, 其中68例(97.1%)患儿在28 d内转阴, 另2例(2.9%)患儿随访修正诊断为...     

EB病毒感染及其相关性疾病

EB病毒(EBV)感染是儿科较为常见的病毒感染性疾病,不同的免疫状态有不同的临床表现。机体感染EBV后,可以产生传染性单核细胞增多症、EBV相关性噬血细胞综合征、慢性活动性EBV感染、X连锁淋巴组织增生性疾病及鼻咽癌、淋巴瘤等。免疫缺陷患者感染EBV后容易发生淋巴组织增殖性疾病。传染性单核细胞增多症是EBV感染最常见的临床表现形式,常合并一系列的并发症。     

基础疾病对脓毒性休克患儿临床特征和预后的影响

目的了解儿童重症监护病房(pediatric intensive care unit,PICU)内不同基础性疾病并发脓毒性休克(septic shock,SS)患儿的临床特征和预后。方法回顾性收集2017年1月1日至2019年12月31日北京儿童医院PICU收治的SS患儿病历资料,按照有无基础疾病、基础疾病种类进行分组,总结不同基础疾病条件下SS的临床特征、预后及病原分布情况。结果研究期间共收治218例SS患儿,总病死率为21.6%(47/218);合并基础疾病者141例[64.7%(141/218)],病死率24.1%(34/141);处于化疗骨髓抑制期的恶性血液病和肿瘤患儿病死率最高(17/45,37.5%),无基础性疾病者病死率最低(13/77,16.9%)。合并基础性疾病的SS患儿以革兰阴性菌感染为主(63.1%,41/65),恶性血液病及肿瘤化疗后骨髓抑制期患儿革兰阴性菌感染最高(80.0%,20/25)。革兰阳性菌感染在无基础疾病组最高(52.1%,25/48)。多器官功能障碍综合征(multiple organ dysfunction syndrome,MODS)在恶性血液病及肿瘤化疗后骨髓抑制期者的发生率最高(95.6%,43/45),无基础疾病组最低(59.7%,46/77)。结论伴发基础疾病,尤其是血液肿瘤的患儿化疗后发生SS时病原体以革兰阴性菌最常见,病死率和MODS发生率高;无基础疾病者以革兰阳性菌最常见,病死率和MODS发生率相对较低。     

中山市儿童EB病毒感染情况分析

目的了解广东省中山市儿童EB病毒(EBV)感染情况及流行特征。方法回顾性分析2011至2013年经实时荧光定量PCR检测外周血中单个核细胞EBV-DNA的疑似EBV感染患儿的临床资料。结果 3 402例疑似EBV感染患儿中,EBV-DNA检出阳性409例,总阳性率为12.0%。其中2011年阳性率为8.1%,2012年为10.4%,2013年为19.5%,三年阳性率差异有统计学意义(P0.05);不同性别患儿间的差异无统计学意义(P0.05);不同年龄组间的差异有统计学意义(P0.05),以学龄前组为最高。EBV感染性疾病以传染性单核细胞增多症为最多(61.6%),其次为呼吸道感染(26.7%)、颈淋巴结炎(3.4%)、血小板减少性紫癜(2.4%)等。在部分EBV感染患儿中,检出MP-IgM阳性79例,CP-Ig M阳性47例,细小病毒B19-IgM阳性20例,单纯疱疹病毒IgM阳性11例,巨细胞病毒IgM阳性10例,风疹病毒Ig M阳性4例。结论中山市儿童EBV-DNA阳性年检出率呈上升趋势,EBV感染后以传染性单核细胞增多症最常见,但可累及多个系统。EBV感染后可能伴有多种病原抗体阳性。     

Prevalence of Epstein-Barr Viral DNA among children at a single hospital in Suzhou,China

ObjectiveThis study aimed to describe the prevalence of Epstein-Barr virus (EBV)-DNA among children in Suzhou, and to explore the association between plasma EBV load and disease diagnosis.MethodsAll children admitted to the Children's Hospital of Soochow University between January 2018 and September 2020 and subjected to the plasma EBV-DNA assay were included. The authors retrospectively collected demographic and discharge diagnostic information of the participants, and ascribed the disease distribution characteristics of children with positive plasma EBV-DNA by age and viral load.ResultsA total of 38,175 patients underwent plasma EBV-DNA PCR assay, of which 2786 (7.3%) had EBV-DNA in their plasma. Children aged 3–4 years had a high prevalence of EBV infection. Plasma EBV positivity was common with infectious mononucleosis (IM, 40.0%), respiratory infection (20.1%), atypical EBV infection (14.2%), acute leukemia (6.4%), hemophagocytic lymphohistiocytosis (HLH, 4.8%), and idiopathic thrombocytopenic purpura (ITP, 2.9%). With increasing age, plasma EBV positivity was more common in children with IM and atypical EBV infection. However, an inverse correlation was observed in children with respiratory infections and ITP. High levels of EBV loads were more likely to occur in HLH, IM, and atypical EBV infection, especially in HLH. However, lower viral loads were found in respiratory infection and acute leukemia.ConclusionsThis is a large sample study that revealed the prevalence of plasma EBV-DNA levels in children of various ages and presenting illnesses.     

761例住院儿童EB病毒感染分析

目的：了解儿童EB病毒（EBV）感染情况,并分析其相关疾病谱,从而为EBV感染及相关疾病的防治提供科学的理论依据。方法：采用real-time PCR法检测2010年8月至2011年7月收治的761例（年龄22 d至14岁）疑似EBV感染儿童血浆中EBV-DNA载量,并对EBV-DNA检查结果及相关疾病进行统计学分析。结果：761例血浆标本中EBV-DNA阳性标本109例,阳性率为14.3%;不同年龄组EBV-DNA阳性检出率差异有统计学意义（P<0.05）,其中婴儿组（<1岁）的阳性检出率最低（P<0.05）;不同季节间阳性检出率差异有统计学意义（P<0.05）,其中夏季阳性检出率高于冬季（P<0.05）。109例阳性标本的EBV-DNA载量范围为2.13～6.69,中位数为3.72。对62例EBV-DNA阳性住院患儿最终临床诊断分析得出,呼吸系统疾病占39%,主要为急性支气管炎、急性上呼吸道感染及急性支气管肺炎。结论：不同年龄组及不同季节间EBV-DNA阳性检出率不同;儿童EBV感染相关疾病以呼吸系统疾病为主;Real-time PCR法检测血浆EBV-DNA有助于临床上EBV感染的早期诊断。     

PCR法检测140例儿童EB病毒DNA的临床分析

目的探讨应用聚合酶链反应技术(PCR)检测EBV-DNA的临床应用及140例EBV阳性病例的临床分析。方法应用PCR和荧光检测技术检测外周血EBV-DNA,并对140例阳性病例的临床特点进行回顾性分析。结果病例年龄1月～12岁,中位年龄3岁,<3岁84例,3～7岁45例,>7岁11例;140例患儿中传染性单核细胞增多症17例(12.1%),呼吸道感染20例(14.3%),另外还有原发性血小板减少性紫癜、肝炎、嗜血细胞综合征、过敏性紫癜等及隐性感染病例;结论PCR法检测EBV-DNA时间短,准确性好,灵敏度高,在EB病毒感染相关疾病诊断中很有价值。     

慢性活动性EB病毒感染患儿外周血单个核细胞EB病毒DNA及感染细胞类型检测的临床意义

目的 研究EB病毒(Epstein-Barr virus,EBV)慢性活动性感染(CAEBV)、急性感染(AEBV)以及正常儿童的外周血单个核细胞(PBMC)的EBV-DNA水平,以及EBV感染细胞类型的差异,探讨其与CAEBV临床表型的关系.方法 收集2004年3月至2008年4月在我院住院的CAEBV患儿10例,AEBV患儿13例,以及正常儿童12例的外周血单个核细胞,应用实时荧光定量PCR法检测EBV-DNA水平,并对EBV-DNA阳性的CAEBV和AEBV及正常儿童,用免疫磁珠法分选各种淋巴细胞后进行EBV编码的RNA-1(EBV encoding RNA-1,EBER-1)探针荧光原位杂交(FISH)确定EBV感染细胞的类型.结果 CAEBV组EBV-DNA载量为[(6.8×107)±(1.1 x 108)]/ml,AEBV组EBV-DNA载量为[(1.3×106)±(1.6×106)]/ml,两组比较差异有统计学意义,CAEBV组PBMC的EBV-DNA水平明显高于AEBV组(P＜0.01);7例CAEBV患儿做细胞分选及FISH后,发现EBV不仅可以引起B细胞感染,而且还引起NK细胞、CD4+和CD8+T细胞不同程度的感染,临床表现为反复或持续的传染性单核细胞增多症(IM)样症状.6例患儿以感染T细胞为主,其中1例以CD8+T细胞感染为主,临床表现除高热,肝脾淋巴结大,伴严重的血液系统一系或三系降低外,还并发了爆发性的致死性T淋巴细胞增殖综合征而死亡.1 例以NK细胞感染为主,临床表现还伴有对蚊虫叮咬高度敏感且IgE高达2500 U/ml.AEBV组7例患儿均显示感染B淋巴细胞,临床表现为可以痊愈的IM.6例正常儿童均为阴性.结论 CAEBV患儿体内存在更多的EBV复制和不同的EBV感染细胞类型,实时荧光定量PCR检测EBV-DNA水平并测定EBV感染的淋巴细胞类型有可能协助CAEBV临床个体化诊治和评估病情进展.

Abstract：

Objective To study the difference in the EBV-DNA level in peripheral blood mononuclear cells (PBMC) and the type of Epstein-Barr virus(EBV)-infected cells in pediatric patients with chronic active EBV(CAEBV) infection,acute EBV infection(AEBV)and healthy children,and to analyze the relationship between the above difference and the clinical manifestation of CAEBV.Method Real-time fluorescent quantitative polymerase chain reaction (PCR) was used to detect the EBV-DNA levels in peripheral blood mononuclear cells (PBMC) in 12 normal children, 10 pediatric patients with CAEBV infection and 13 pediatric patients with AEBV infection in our hospital between March 2004 and April 2008. Immunomagnetic bead cell fractionation and fluorescent in situ hybridization(FISH) by EBV encoding RNA-1(EBER-1) probe were used in the healthy children, EBV-DNA positive CAEBV patients and AEBV patients to detect the type of EBV-infected cells.Result The average EBV-DNA level in CAEBV patients'PBMC was(6.8×107±1.1×108)copies/ml, while the average EBV-DNA level of AEBV patients' PBMC was(1.3×106±1.6×106)copies/ml.The average EBV-DNA level of CAEBV infected patients' PBMC was significantly higher than that of AEBV infected patients' PBMC(P＜0.01).The cell fractionation and FISH in seven CAEBV patients showed that EBV in CAEBV patients infected not only B cells,but NK cells and CD4+ and CD8+ T cells to different degree, and these patients presented recurrent and persistent infectious mononucleosis(IM)-like symptoms.In 6 CAEBV patients infection mainly occurred to T cells, in one case,infection occurred mainly in CD8+T cells, and the patient died from fulminant and deadly T lymphocytes proliferative syndrome except presenting firstly high fever, enlargment of the liver, spleen, lymphnode and the severe decrease of one or three kinds of blood cells. In 1 CAEBV patient the infection was mainly found in NK cells, who presented with hypersensitivity to mosquito biting and high IgE level (2500 U/ml).But EBV in seven AEBV patients infection was found only in B cells who presented with only IM for one time and no EBV-infected PBMC were found in the remaining 6 healthy children. Conclusion There are much more EBV replications and different EBV-infected cell types in CAEBV patients. Detection of EBV-DNA level by real-time fluorescent quantitative PCR and the detection of the type of EBV-infected cells may help in diagnosis, treatment and development evaluation of children with CAEBV infection.     

儿童再生障碍性贫血病毒病因学的初步研究

用ＰＣＲ和ＥＬＩＳＡ技术对３８例ＡＡ患儿的外周血进行了１９、ＥＢＶ、ＣＭＶ及ＨＢＶ系列检测。     

Epstein-Barr virus DNA load and seroconversion in pediatric renal transplantation with tacrolimus immunosuppression

EBV infection is one of major complications arising in pediatric patients who have undergone renal transplantation. A strong correlation between the grade of immunosuppression and the development of PTLD, one of the most severe EBV-associated diseases, has been recognized. In this study, we monitored the serologic profile in conjunction with peripheral blood EBV-DNA load of 32 children who underwent renal transplantation with tacrolimus as an immunosuppressant. Six patients were EBV-seronegative (EBV-) before the transplantation, and the mean DNA load in the EBV- group was significantly higher than that in the EBV-seropositive (EBV+) group. Seroconversion occurred in five of these patients in a mean period of 22 weeks after the transplantation. The EBV-DNA load in the EBV+ group was maintained at a low level for a year, whereas it increased rapidly to over 1 x 10(5) copies/mL in two patients in the EBV- group three to seven months after the transplantation, which corresponds to the timing of seroconversion, and one of them developed PTLD. These observations suggest that the close monitoring of the EBV-DNA load, along with longitudinal observation of seroconversion, is essential in pediatric renal transplantation, particularly for younger children who are more likely to be EVB-.     

91例EB病毒相关疾病儿童血浆EB病毒DNA的检测

目的了解EB病毒(EBV)感染患儿外周血血浆中游离EBVDNA的拷贝数,确定EBV原发感染后外周血血浆中EBV游离DNA的拷贝数与发病天数及病情轻重的关系。方法应用荧光定量PCR方法,测定73例EBV原发感染和18例EBV相关重症疾病患儿外周血血浆中EBV游离DNA。结果①原发EBV感染患儿外周血血浆中EBV游离DNA随发病天数呈下降趋势,发病2周后很难检测到。②EBV相关重症疾病组患儿外周血血浆中EBV游离DNA阳性率明显高于原发EBV感染组,差异有显著性(89%vs16%,P<0.05)。结论原发EBV感染后随病程天数的增加,病毒复制水平逐渐下降。血浆中EBV游离DNA检测对评价EBV相关疾病的严重程度有一定参考价值。     

宏基因组二代测序在伴有中枢神经系统受累儿童噬血细胞综合征中的应用价值

目的 探究宏基因组二代测序（metagenomic next-generation sequencing,mNGS）技术检测伴有中枢神经系统受累的噬血细胞综合征患儿颅内EB病毒（Epstein-Barr virus,EBV）感染的应用价值及临床意义。 方法 回顾性分析30例伴有中枢神经系统受累的噬血细胞综合征患儿的脑脊液mNGS结果,与脑脊液EBV-DNA定性检测、血清EBV抗体谱检测结果进行比较,以治疗前后血清EBV-DNA拷贝数变化反映针对性治疗效果。 结果 脑脊液mNGS EBV检测阳性率为100%（30/30）,高于脑脊液EBV-DNA定性检测阳性率（10%,3/30;P<0.001）,与血清EBV抗体谱检测阳性率（93%,28/30）比较差异无统计学意义（P>0.05）。中位mNGS EBV检出序列数为2 400,治疗前血清EBV-DNA拷贝数与EBV检出序列数呈中度正相关（r_s=0.693,P<0.001）。多元线性回归分析结果显示,治疗前血清EBV-DNA拷贝数越高,脑脊液mNGS EBV检出序列数越高（P<0.05）。 结论 EBV相关噬血细胞综合征容易诱发EBV感染引起的病毒性脑炎,mNGS可以显著提高脑脊液EBV检测的阳性率,协助临床诊断。     

91例EB病毒相关疾病儿童血浆EB病毒DNA的检测

目的:了解EB病毒（EBV）感染患儿外周血血浆中游离EBV DNA的拷贝数,确定EBV原发感染后外周血血浆中EBV游离DNA的拷贝数与发病天数及病情轻重的关系。方法:应用荧光定量PCR方法,测定73例EBV原发感染和18例EBV相关重症疾病患儿外周血血浆中EBV游离DNA。结果:①原发EBV感染患儿外周血血浆中EBV游离DNA随发病天数呈下降趋势,发病2周后很难检测到。②EBV相关重症疾病组患儿外周血血浆中EBV游离DNA阳性率明显高于原发EBV感染组,差异有显著性(89% vs 16%, P＜0.05）。结论: 原发EBV感染后随病程天数的增加,病毒复制水平逐渐下降。血浆中EBV游离DNA检测对评价EBV相关疾病的严重程度有一定参考价值。［中国当代儿科杂志,2009,11（11）：897-900］     

p53 protein expression does not correlate with EBV status in childhood B non-Hodgkin lymphomas

BACKGROUND: The p53 tumor suppressor gene is affected in a wide range of human cancers, including hematological malignancies. This gene encodes a nuclear phosphoprotein p53, which plays a key role in cell cycle arrest, induction of apoptosis, and DNA repair. Mutations of the p53 gene often lead to the accumulation of the mutated protein in the nucleus of neoplastic cells. However, p53 protein expression is frequently detected in non-Hodgkin lymphomas (NHL) without any correlation with p53 mutations. This discordance suggests the existence of other mechanisms to stabilize the p53 protein, including binding of p53 protein to viral proteins. p53 protein has been shown to bind to proteins encoded by the Epstein-Barr virus (EBV). PROCEDURE: The aim of this study was to analyze p53 expression in childhood B-NHL and correlate its expression in the absence of p53 mutations with EBV in order to investigate a possible involvement of EBV in p53 stabilization. DESIGNS AND METHODS: Tumor specimens from 35 children with B-NHL were analyzed by immunohistochemistry (IHC) with the DO7 monoclonal antibody, which recognizes an epitope at N-terminus of p53 protein and reacts with wild type and mutant proteins. To detect p53 mutations, PCR/SSCP and sequencing were performed. EBV status was determinated using a specific PCR technique. RESULTS: The overall frequency of p53 immunostaining positivity was 45% (16 of 35). p53 mutations were detected in nine patients (25.6%). p53 immunoreactivity was observed in all cases with mutations. Additionally, we identified 7 p53 positive cases among 26 tumors without mutations. EBV DNA was detected in 24 of 35 cases. Four patients with p53 expression dissociated from mutation were EBV positive. No statistically significant association was found between p53 expression and EBV cases despite the exclusion of those patients in which p53 expression was related with p53 mutations (P = 0.28 and 0.54, respectively). CONCLUSIONS: Our results suggested that in childhood B-NHL, the expression of p53 dissociated from mutations could not be related to EBV infection. Further studies with larger patient sets will be necessary to determinate if EBV-encoded protein may play a role for nuclear accumulation of p53 protein.