EB病毒抗体检测在不同年龄壮族鼻咽癌患者中的应用

目的探讨桂西地区壮族鼻咽癌患者EB病毒各年龄段Rta/IgG、VCA/IgA、VCA/IgG及Zta/IgG抗体的rA值和阳性率与年龄的关系。方法收集140 例未经治疗的鼻咽癌患者和280例健康人的血清,用酶联免疫吸附法（ELISA）检测Rta/IgG、VCA/IgA、VCA/IgG及Zta/IgG抗体,分别计算各年龄段的抗体水平及阳性率并进行统计学分析。结果鼻咽癌患者各年龄段VCA/IgA抗体rA值和阳性率差异均有统计学意义 (P<0.05)。在50岁以后年龄段的患者与健康人Rta/IgG、VCA/IgA抗体阳性率比较差异有统计学意义 (P<0.05)。在50岁以后的年龄段患者与健康人Zta/IgG抗体阳性率比较差异无统计学意义 (P>0.05)。结论壮族鼻咽癌患者与健康人EB病毒Rta/IgG、VCA/IgA及Zta/IgG抗体水平和阳性率比较存在年龄上的差异。在鼻咽癌的临床诊断和高危人群筛查中有必要根据不同年龄段的人群界定不同的阳性临界值。     

EBVDNA和EBV抗体指标早期诊断鼻咽癌方案的筛选

目的:探讨EB病毒DNA及EB病毒抗体在鼻咽癌联合诊断中的应用价值.方法:对81例未经治疗的病理确诊鼻咽癌怠者和89例健康体检者分别采用实时荧光定量PCR测定EB病毒DNA,计算病毒拷贝数,采用间接免疫酶法测定EBV VCA/IgA和EMIgA,采用酶联吸附试验(ELISA法)检测EBNA1/IgG、EBNA1/IgA以及ZTA/IgG.通过ROC曲线评价各指标单独和联合诊断鼻咽癌的效果.结果:分析发现VCA/IgA、EBV DNA和EBNA1/IgA 3个指标具有较高的灵敏度,分别为95.06%、83.95%和81.63%.而EA/IgA、EBNA1/IgG和ZTA/IgG则具有较高的特异度,分别为94.38%、95.51%和89.89%.EBNA1/IgA与VCM/IgA并联时灵敏度为100%,特异度为84.27%.结论:EB-NA1/IgA与VCA/IgA并联时灵敏度和特异度都在比较高的水平,可作为鼻咽癌早期筛查的选择方案.     

鼻咽癌高发区居民EB病毒感染流行病学研究

分析广东省鼻咽癌高发区30~59岁16 355人EB病毒EBNA1/IgA和VCA/IgA抗体在不同性别、年龄人群中的分布及患鼻咽癌概率情况。方法：应用ELISA法检测EBNA1/IgA和VCA/IgA抗体,计算抗体阳性率和鼻咽癌风险概率,从不同层面分析血清EB病毒抗体特征。结果：人群EBNA1/IgA阳性率为4.04%,男、女分别为5.11%和3.23%,除30～39岁年龄段以外,EBNA1/IgA阳性率均显示男性高于女性（P<0.05）。人群VCA/IgA阳性率为5.84%,男女性分别为5.97%和5.74%,35～59岁年龄段VCA/IgA阳性率表现与性别无关（P>0.05）。男性EBNA1/IgA和VCA/IgA阳性率与年龄呈正相关（P<0.05）,各年龄段抗体阳性率分布与鼻咽癌发病年龄趋势基本吻合,尤其是EBNA1/IgA与年龄显示了较好的一致性。女性中以上指标未见随年龄增长而上升。结论：鼻咽癌高发区人群EBNA1/IgA抗体分布存在性别上的差异。男性人群的EBNA1/IgA和VCA/IgA阳性率随着年龄增长逐渐上升,女性以上指标与年龄无显著相关。EBNA1/IgA抗体阳性率高峰年龄段与鼻咽癌高发年龄段具有较好的一致性,有望成为鼻咽癌筛查的理想指标。     

血清EB病毒抗体EBNA1 IgA检测临界值探讨

目的:通过对健康人群与鼻咽癌患者EB病毒抗体EBNA1 IgA水平的分析,探讨在鼻咽癌高发区应用该抗体作为鼻咽癌血清学指标时临界值的确定.方法:采用ELISA法检测780例健康人和104例鼻咽癌患者血清EB病毒EBNA1 IgA,根据灵敏度和特异度曲线分别选择灵敏度、特异度在95%所对应的rOD值作为阴性临界值和阳性临界值,根据该临界值将人群划分成3个不同等级,rOD≥1.85为阳性,1.85＞rOD≥1.10为可疑阳性,rOD＜1.10为阴性.结果:健康人群EBNA1 IgA的均值是0.850±0.637,鼻咽癌患者为2.241±0.875.健康人群中EBNA1 IgA阴性、可疑阳性和阳性人群分别占75.13%、17.44%和7.44%,而鼻咽癌患者则分别是4.81%,17.31%,77.88%.结论:鼻咽癌高发区人群血清EBNA1 IgA的rOD值离散程度较大.以EBNA1 IgA作为鼻咽癌血清学诊断指标,可根据灵敏度和特异度确定阳性、可疑阳性和阴性3个人群,以利于临床作出对鼻咽癌的辅助诊断.     

鼻咽癌患者配偶及子女血清EB病毒IgA/VCA抗体水平分析

目的:通过对鼻咽癌患者及其配偶和子女的血清中EB病毒IgA/VCA抗体水平进行分析,进一步了解鼻咽癌患者家属患鼻咽癌的风险情况.方法:选择鼻咽癌患者及其配偶和子女各317例,同一地区正常人群413例,用免疫酶法检测血清中EB病毒IgA/VCA抗体.结果:鼻咽癌组、配偶组、子女组及对照组的IgA/VCA抗体阳性率分别97.2%、14.2%、19.9%及3.1%,鼻咽癌组显著高于其它3组,配偶组和子女组也明显高于对照组(P＜0.001).经Ridit分析显示,鼻咽癌组、配偶组、子女组与对照组的平均Ridit值分别为0.860、0.404、0.424及0.356.鼻咽癌组的IgA/VCA抗体滴度水平显著高于其它3组,配偶组和子女组的IgA/VCA抗体滴度水平也明显高于对照组(P＜0.001).鼻咽癌患者的配偶与子女IgA/VCA阳性感染的OR值分别为5.09和7.63.配偶组与子女组的IgA/VCA抗体阳性率及抗体滴度水平比较差异均无统计学意义(P＞0.05).结论:EB病毒感染IgA/VCA抗体阳性具有家族聚集性,鼻咽癌患者的配偶及子女EB病毒被再激活的机会要比正常人高,对该高危人群应结合临床密切随访及观察.以便早期发现鼻咽癌.     

Logistic回归模型在早期鼻咽癌多指标联合辅助诊断实验的应用

背景与目的:目前用于鼻咽癌辅助诊断的指标较多,但大部分早期诊断的效果较差.如何联合多个指标进行辅助诊断是近年研究的重点.本研究旨在评价鼻咽癌常用诊断指标VCA/IgA、EA/IgA、EBV DNA、EBNA1/IgA、EBNAl/IgG和ZTA/IgG的诊断价值.并采用logistic回归筛选优化的联合诊断组合.方法:对8l例未经治疗的病理确诊鼻咽癌患者和89例健康体检者分别采用实时荧光定量PCR法测定EB病毒DNA,计算病毒拷贝数,采用间接免疫酶法测定EBV、VCA/IgA和EA/IgA,采用酶联吸附试验(ELISA法)检测EBNA1/IgG、EBNA1/IgA以及ZTA/IgG.通过ROC曲线评价各指标单独和联合诊断鼻咽癌的效果.用logistic回归综合两指标信息,根据ROC曲线确定诊断界值,以增加鼻咽癌诊断的准确性.结果:logistic回归方法的诊断敏感度和特异度比常规的并联实验均有提高.筛选的优化组合方案为EBV DNA+EBNA1/IgA和VCA/IgA+EBNAI/IgA,两组合的并联试验敏感度和特异度分别为0.96、0.82和1.00、0.84,而logistic回归预测概率为诊断指标.ROC曲线确定界值时.两组合的灵敏度和特异度分别为1.00、0.87和0.98、0.88.结论:多指标联合建立logistic回归模型,利用预测概率进行辅助诊断能够达到同时提高灵敏度和特异度的效果,可供临床参考.     

鼻咽癌病人多种抗EB病毒抗体反应的相关性与同份血清多指标检测的应用价值

本文收集广州、中山县等鼻咽癌高发区的鼻咽癌病人和正常人的血清样本,应用免疫荧光和免疫酶标技术检测EB病毒壳抗原(VCA)、早期抗原(EA)的IgA或IgG型抗体以及核抗原(EBNA)抗体水平。 VCA与EA两者无论是IgA或IgG型抗体都有高度相关关系,而EBNA抗体与VCA或EA抗体的出现无相关性。VCA与EA的相关系数(γ)在0.265～0.731之间,表示两者有作常显著的相关关系(P值<0.01或0.001)。相反地,当EBNA抗体显著升高时,VCA和EA抗体可以不出现或抗体水平很低,EBNA抗体滴度与VCA或EA抗体滴度的增加无平行关系,二者间统计学上无显著意义。 上述检测结果表明:多种抗EB病毒抗体的检测方法的综合应用,不仅提高了鼻咽癌早期诊断的阳性率,而且可以降低由单一种EBV抗体检测方法所造成的假阴性。 本文还讨论了淋巴细胞亚型(B或T)与EBNA、VCA和EA三种抗体的相互关系,特别是T细胞功能与EBNA抗体水平的关系。     

鼻咽癌血清学检测中的复合阳性判断方法及其应用

EB病毒 (Epstein-Barr virus, EBV)与鼻咽癌的发生和发展密切相关 , 95% 以上的鼻咽癌血清都含有与 EB病毒壳抗原 (virus capscule antigen,VCA)相关的 IgA抗体 (IgA/VCA抗体 ). IgA/VCA(+ )人群被称为鼻咽癌的高危群体 [1]. 根据闵华庆等 [2]对 12万人的调查结果 , 在占总数 8.15% 的 IgA/VCA(+ )人群中 ,鼻咽癌的检出率比 IgA/VCA(- )人群高 40倍 . 尽管如此 , 在 IgA/VCA(+ )人群中鼻咽癌的阳性检出率仍然很低 , 只有 1.21% . 因此 , 有必要在 IgA /VCA(+ )人群中继续探索能提高鼻咽癌检出率和诊断准确性的方法 , 这也是关系到在鼻咽癌高发区如何提高血清学筛查效果的关键之一 . 1997年黄腾波等在广东四会鼻咽癌高发区对 IgA/VCA(+ )人群进行监测研究 , 提出了血清学与临床体检相结合的综合筛查方案 [3]. 当时检测 EBV早期复合抗原 (EBV-EA)相关 IgG抗体的酶联免疫吸附 (ELISA)法 (IgG/EA ELISA)尚未建立 . ELISA法与间接免疫酶染色 (IEA)检测 EBV-EA IgG抗体滴度的方法 (IgG/EA IEA)比较 , 在敏感性相同 (88.3% )的前提下 , 具有特异性 (98.3% )明显优于后者 (53.2% )的特点 [4] . 在特异性为 97.7% 的前提下 , 用 ELISA法检测鼻咽癌血清的敏感性 (89.2% )也显著高于用 IEA法测定 EBV-EA IgA抗体滴度 (IgA/EA IEA)的敏感性 (65.5% )[5].     

鼻咽癌患者一级亲属不同性别人群EB病毒抗体和患病概率分析

目的:通过对散发性鼻咽癌患者一级亲属EB病毒感染和患病概率分析,了解遗传和性别因素对EB病毒抗体水平和患鼻咽癌概率的影响.方法:用ELISA法检测散发性鼻咽癌患者一级亲属(亲属组)和一般人群(对照组)血清EB病毒VCA/IgA和EBNA1/IgA抗体,统计不同人群抗体阳性率、鼻咽癌风险概率和癌检出率.结果:亲属组和对照组的VCA/IgA阳性率分别为6.78％和5.83％,EBNA1/IgA阳性率分别为6.10％和4.00％,抗体阳性率组间差异均无统计学意义(P＞0.05),其中男亲属组的阳性率高于男对照组(P＜0.05).男亲属组的鼻咽癌高危率大于男对照组(P＜0.05).在亲属组与对照组,VCA/IgA阳性率显示与性别无关(P＞0.05),而EBNA1/IgA阳性率和鼻咽癌高危率均表现为男性高于女性,P＜0.05.男亲属组癌检出率大于男对照组,P＜0.05;男对照组癌检出率大于女对照组,P＜0.05.结论:散发性鼻咽癌患者男性一级亲属EBNA1/IgA抗体阳性率和患鼻咽癌风险较一般男性高,而女性亲属以上指标与一般女性无差异.散发性鼻咽癌一级亲属中男性患鼻咽癌的风险高于女性.男性一级亲属鼻咽癌患病率高于一般人群,再次提示由血缘关系反映的遗传因素是鼻咽癌发病的主要原因.     

以血清EB病毒抗体谱评估患鼻咽癌危险度的研究

目的：比较鼻咽癌患者与健康人群血清EB病毒抗体水平,评估患鼻咽癌的危险度。方法：收集珠江口地区121例鼻咽癌患者和332例健康人血清。采用酶联吸附试验（ELISA）检测血清中EBNA 1/IgA、EBNA 1／IgG和zta/IgG,以免疫酶法或免疫荧光法检测VCA／IgA。结果：EBNA 1／IgA、EBNA 1／IgG和zta/IgG的敏感度分别为85％、83％和79％;三者组合的敏感度最高,为92％。EBNA 1/IgA、EBNA 1／IgG和zta/IgG的特异度分别为86％、86％和80％;三者组合的特异度最高,为93％。根据优势比水平,可将患鼻咽癌的危险度分为低危险、中危险和高危险3个层次。93％的健康人是低危险的,优势比为0．0－0．3;0．4％的健康人是高危险的,优势比为137．9。结论：ELISA在诊断鼻咽癌的结果判断上更具客观性,较传统免疫酶法好;EBNA 1／IgA、EBNA 1／IgG和zta/IgG的联合应用可以评估人群患鼻咽癌的危险度。     

Diagnostic Significance of Combined Detection of Epstein-Barr Virus Antibodies,VCA/IgA,EA/IgA,Rta/IgG and EBNA1/IgA for Nasopharyngeal Carcinoma

The objective of this study was to investigate the diagnostic significance of EBV antibody combined detectionfor nasopharyngeal carcinoma (NPC) in a high incidence region of southern China. Two hundred and elevenuntreated NPC patients, 203 non-NPC ENT patients, and 210 healthy controls were recruited for the study. Thetiters of VCA/IgA and EA/IgA were assessed by immunoenzyme assay, and the levels of Rta/IgG and EBNA1/IgAwere determined by enzyme-linked immunosorbent assay. The levels of VCA/IgA, EA/IgA, Rta/IgG and EBNA1/IgA demonstrated no association with gender or age (p>0.05). The receiver operating characteristic curve andthe area under the curve were used to evaluate the diagnostic value. The sensitivity of VCA/IgA (98.1%) andthe specificity of EA/IgA (98.5%) were the highest. When a logistic regression model was used to combine theresults from multiple antibodies to increase the accuracy, the combination of VCA/IgA+Rta/IgG, whose areaunder the curve was 0.99, had the highest diagnostic efficiency, and its sensitivity, specificity and Youden indexwere 94.8%, 98.0% and 0.93 respectively. The data suggest that the combination of VCA/IgA+Rta/IgG may bemost suitable for NPC serodiagnosis.     

EBV 抗体和EBV-DNA 在鼻咽癌诊断及分期的研究*

目的：评估EBNA1/IgA 、Zta/IgA 、VCA/IgA 和EBV-DNA对不同分期鼻咽癌的诊断效能,探讨各指标阳性率与鼻咽癌分期的关系。方法：收集2010年3 月至2015年9 月中山大学附属中山医院收治的初诊鼻咽癌患者152 例,健康体检者675 例。采用酶联免疫吸附法（ELISA）检测血清EBNA1/IgA 、Zta/IgA 和VCA/IgA 抗体ROD 值,荧光定量PCR （fluorescence quantitative PCR,FQ-PCR ）检测血浆EBV-DNA水平。比较单独和联合应用EBV 标记物对各期鼻咽癌的诊断效能,同时分析各指标阳性率与鼻咽癌分期的关系。结果：鼻咽癌患者EBNA1/IgA 、Zta/IgA 、VCA/IgA 和EBV-DNA阳性率显著高于健康体检者（P < 0.01）。 EBNA1/IgA 在早期鼻咽癌表达相对较高,灵敏度为77.8% ,而EBV-DNA在晚期鼻咽癌的灵敏度最高为88.8% ,两者特异度均在96% 以上。联合检测中EBNA1/IgA 并联EBV-DNA检测的灵敏度为92.1%（早期为82.5% 、晚期为98.9%）,特异度为96.9% 。EBV-DNA阳性率与鼻咽癌临床分期和N 分期呈正相关,Zta/IgA 阳性率与N 分期呈正相关（P < 0.01）。 结论：在无症状人群中进行鼻咽癌筛查,单项指标首选EBNA1/IgA 。晚期患者的辅助诊断则推荐EBV-DNA。两者并联检测可进一步提高鼻咽癌诊断效能。EBV-DNA是鼻咽癌分期和病情监测的重要指标,Zta/IgA 可间接反映淋巴结转移情况,有望对患者病情评估起到参考作用。     

Significance of Plasma IgA and IgG Antibodies to Epstein-Barr Virus Early and Viral Capsid Antigens in Thai Nasopharyngeal Carcinoma

Epstein-Barr virus (EBV) is an important causal factor of human nasopharyngeal carcinoma (NPC). High levels ‍of serum IgA and IgG antibodies to EBV early and viral capsid antigens (IgA/EA, IgA/VCA, IgG/EA and IgG/VCA) ‍have been reported in NPC patients. Since specific serum IgA/EA, IgA/VCA and IgG/EA are claimed to be useful ‍serological markers for NPC. In order to evaluate whether plasma IgA/EA, IgA/VCA, IgG/EA and IgG/VCA antibody ‍levels are useful markers for diagnosis and prognosis of Thai NPC, we examined the prevalence of these antibodies ‍in 79 NPC patients, and 127 age-matched controls (47 healthy subjects (HS), 32 cases of other disease (OD) and 48 ‍cases of other cancer (OC)) by using an indirect immunofluorescence assay. The prevalence of plasma IgA/EA, IgA/ ‍VCA, and IgG/EA in NPC patients (55.7, 68.4 and 68.4%) was significantly higher than in the HS (0.0, 0.0 and ‍20.5%,), OD (0.0, 0.0 and 3.1%) and OC (0.0, 0.0 and 20.8%) groups (p<0.05). The prevalence of plasma IgG/VCA ‍in NPC patients (93.7%) was significantly different from those for the OD and OC groups (71.9 and 43.8%) but not ‍for the HS group (89.4%). In NPC patients, the geometric mean titers (GMT) of plasma IgA/EA, IgA/VCA and IgG/ ‍EA were increased with an advanced clinical stage of disease but not IgG/VCA. In contrast, GMT of IgG/VCA was ‍increased with aggressive type of disease (histological type) but not IgA/EA, IgA/VCA, and IgG/VCA. The results of ‍our study suggest that plasma IgA/EA, IgA/VCA and IgG/EA antibodies may be useful markers for diagnosis and ‍assessing prognosis of Thai NPC. ‍     

Antibody response to Epstein-Barr virus Rta protein in patients with nasopharyngeal carcinoma: a new serologic parameter for diagnosis.

BACKGROUND: Nasopharyngeal carcinoma (NPC) is associated closely with Epstein-Barr virus (EBV). The authors previously reported that an EBV immediate-early gene, BRLF1, was expressed frequently in NPC tumors, and a significant elevation in immunoglobulin G (IgG) antibodies directed against BRLF1 gene product Rta was detected in NPC sera by a radioactive immunoprecipitation assay. To simplify and to make the detection more quantitative, an enzyme-linked immnunosorbent assay (ELISA) was developed in this study. METHODS: Antigen domains of Rta were identified further using an immunoprecipitation assay. Two glutathione-S-transferase (GST) recombinant Rta fragments (R150-GST and R185-GST) were prepared subsequently and were used as antigens in the ELISA. Serum samples derived from 51 patients with NPC patients, 115 non-NPC ENT patients, and 47 healthy volunteers were examined for the presence of antibodies directed against Rta. RESULTS: Among the patients with NPC, 74.5% showed a positive IgG response to R150-GST, and 62.7% showed a positive IgG response to R185-GST, with 80.4% positive for either fragment. In contrast, the reactions were positive in only 8.5% of healthy volunteers and 13.0% of control patients. When using a mixture of the two recombinant Rta proteins as coating antigens, the IgG positive responses were 82.3%, 10.6%, and 14.8%, respectively, in patients with NPC, healthy volunteers, and control patients. It is noteworthy that 51.0% of the NPC sera showed a positive immunoglobulin A (IgA) response, with none of the control patients showing obvious reactivity. Both the IgG response and the IgA response to Rta protein in patients with NPC were correlated with the IgA response to EBV early antigens and virus capsid antigens, the classic serologic markers used to diagnose NPC. CONCLUSIONS: The ELISA method described for the detection of IgG antibodies directed against recombinant Rta proteins is simple and reliable and may be useful as a serologic parameter for the screening and diagnosis of patients with NPC.     

Increased incidence of IgA antibodies to the Epstein-Barr virus-associated viral capsid antigen and early antigens in patients with chronic lymphocytic leukemia

Antibody titers to Epstein-Barr virus (EBV)-associated early antigens (EA) and the viral capsid antigen (VCA) were determined by ELISA on 263 sera obtained from healthy donors, patients with Hodgkin's disease (HD), non-Hodgkin lymphomas (NHL), infectious mononucleosis (IM), Burkitt's lymphoma (BL), and nasopharyngeal carcinoma (NPC). As expected, most lymphoma patients showed markedly elevated anti-VCA IgG and anti-EA IgG antibody titers. Only one patient in the NHL group (n = 56) consisting of patients with lymphomas other than chronic lymphocytic leukemia (CLL) and hairy-cell leukemia (HCL), and 3 patients with HCL (n = 19) had high antibody titers of the IgA class to VCA and EA. Seventeen out of 48 patients (36%) with CLL had high IgA anti-VCA titers and 10 of these sera (21%) also contained IgA anti-EA. The geometric mean titer (GMT) of IgA anti-VCA was 2,510, the GMT of IgA anti-EA was 780. These antibody titers were about 10 times lower than the corresponding GMT of the NPC patients investigated in this study. The elevated IgG and IgA antibody titers to VCA and EA in CLL and HCL patients seem to reflect an immunodeficiency secondary to the malignant disease leading to reactivation of latent EBV infection. The possibility that at least some of these B-cell lymphomas are associated with EBV cannot be excluded.