鼻咽癌围放疗期EB病毒IgA抗体水平的变化

目的:探讨鼻咽癌围放疗期EB病毒免疫球蛋白A（IgA）抗体水平变化的价值。方法:选自我院于2014年4月至2017年3月期间收治的鼻咽癌患者61例;另选自我院于2014年4月至2017年3月行EBV血清学检测的50例健康者作为对照组。空腹采集外周静脉血,分离血清,采用ELISA法测定衣壳抗原IgA（VCA-IgA）和早期抗原IgA（EA-IgA）水平。比较两组VCA-IgA和EA-IgA抗体水平,鼻咽癌患者放疗前后VCA-IgA和EA-IgA抗体水平和阳性率,及VCA-IgA和EA-IgA联合诊断灵敏度和特异度。结果:观察组VCA-IgA和EA-IgA抗体水平高于对照组,且有统计学差异（P<0.05）;鼻咽癌患者放疗后VCA-IgA和EA-IgA抗体水平低于放疗前,且有统计学差异（P<0.05）;鼻咽癌患者放疗后VCA-IgA和EA-IgA阳性率低于放疗前,且有统计学差异（P<0.05）;鼻咽癌患者VCA-IgA和EA-IgA联合诊断灵敏度和特异度高于VCA-IgA和EA-IgA单项诊断。结论:VCA-IgA和EA-IgA用于鼻咽癌患者治疗效果评估具有重要价值,且联合诊断灵敏度和特异度较高,具有重要研究意义。     

联合检测EB病毒不同抗体及EB病毒DNA在鼻咽癌血清学诊断中的价值

目的:评价EB病毒(EBV)抗衣壳抗原VCA-IgA抗体、抗早期抗原EA-IgA抗体、抗立即早期抗原Rta-IgG抗体和EBV-DNA检测在鼻咽癌诊断中的价值。方法:收集160例初治鼻咽癌,133例症状相似的非鼻咽癌患者和163名健康体检者的血清和血浆。采用酶联免疫法检测血清VCA-IgA、EA-IgA和Rta-IgG抗体的水平,用实时荧光定量PCR检测血浆EBV-DNA的相对含量。按鼻咽癌2008临床TNM分期法进行分期,计算不同分组、各临床分期以及鼻咽癌治疗前后的各抗体阳性率、抗体水平以及EB-DNA的检测结果并进行数据分析。结果:鼻咽癌组VCA-IgA、EA-IgA、Rta-IgG和EBV-DNA阳性率均高于鼻咽相关疾病组及健康对照组（均P<0.05）。血清VCA-IgA和EA-IgA结果相对A值（即rA或S/CO值）在Ⅰ、Ⅱ期低于Ⅲ、Ⅳ期,差异有统计学意义(P<0.05),但阳性率在各期间比较差异无统计学意义(P>0.05);Ⅰ、Ⅱ期患者Rta-IgG的rA值和阳性率明显低于Ⅲ、Ⅳ期患者(P<0.05);血浆中EBV-DNA阳性率及EBV-DNA中位数水平随着临床分期的升高而增高,Ⅰ、Ⅱ期与Ⅲ、Ⅳ期比较差异有统计学意义（P<0.05）。治疗有效（CR+ PR）患者的EBV-DNA含量明显低于治疗前水平（P<0.05）。结论:VCA-IgA、EA-IgA、Rta-IgG、EBV-DNA检测有助于鼻咽癌的辅助诊断、临床分期预测及疗效评估。     

鼻咽癌病人血清中EB病毒的IgA,IgG抗体反应

本文应用免疫荧光法检测了294例鼻咽癌、118例鼻咽粘膜病变和40例正常人血清的VCA-IgA、IgG,EA-IgA,IgG。89.8％和26.7％鼻咽癌患者为VCA-IgA和EA-IgA抗体阳性,抗体的几何平均滴度为15.21和1.9。只有7.5％正常人和15.3％鼻咽粘膜病变者有低滴度的VCA-IgA抗体,而EA-IgA抗体,正常人全部阴性,粘膜病变者有2.5％阳性(GMT为1.30)。这些结果表明IgA抗体的出现是鼻咽癌EA病毒血清学的非常特异的现象,其中EA-IgA的阳性率和GMT均较低,但具有更高的特异性。所有被检对象的血清均为VCA-IgG阳性,但鼻咽癌组抗体的GMT(107.67)显著地高于正常人组(64.98)和粘膜病变组(26.20)。EA-IgG在各组中亦有显著差异,提示抗EA抗体也是鼻咽癌的特异现象。 鼻咽癌患者晚期EBV抗体水平显著高于早期。但抗体水平与原发灶大小无关,与淋巴结转移程度有关。在鼻咽癌患者体内,IgA和IgG抗体水平的升高是一致的。 本文还比较了免疫荧光法和免疫酶法在测定VCA-IgA的结果。免疫酶法检出抗体的阳性率和GMT稍高于前者,但无统计学上的差异,两法的特异性也是一致的。     

联合检测EB病毒相关抗体和抗原对诊断鼻咽癌的价值

背景与目的：近年来随着分子生物学的发展,提供了多项EB病毒（Epstein-Barrvirus）相关的检测指标。本研充通过同时检测EB病毒VCA-IgA、EA、IgA、EBV-特异性DNA酶（EBV-DNase）抗体、EB病毒DNA（EBV-DNA）,评价联合检测对诊断鼻咽癌的价值。方法：收集160例治疗前的鼻咽癌患者和76例健康成人的血清和血浆,应用免疫酶染色法检测血清VCA-IgA、EA-IgA：用正丁酸与巴豆油激发Raji细胞方法检测EBV-DNase抗体：用实时荧光定量聚合酶链反应（real-timefluoresc,eneequantitativePCR,RQ．PCR）分析血浆EBV-DNA,评什其在诊断鼻咽癌中的价值。结果：单项检测时对鼻咽癌患者诊断的敏感性和特异性分别为：VCA-IgA90．0％、89．5％,EA-IgA75．0％、94．7％,EBV-DNase抗体76．3％、90．8％,EBV-DNA68．8％、88．2％。联合检测的敏感性和特异性分别为98．8％和84．2％。VCA-IgA、EA-IgA阳性率与临床分期无关（P〉0．05）,各临床分期中EBV-DNase抗体阳性率、EBV-DNA水平与阳性率的差异有统计学意义（P〈0．05）。结论：VCA-IgA、EA-IgA、EBV-DNase和EBV-DNA单项检测时VCA-IgA敏感性最高,EA-IgA特异性最好;四项指标联合检测可提高鼻咽癌诊断的敏感性和准确度。EBV-DNase抗体、EBV-DNA有助于评估鼻咽癌病程和协助判断临床分期。     

提高EB病毒血清学在诊断鼻咽癌中的效益

目的 探讨同时应用VCA-IgA,EA-IgA和EA-IgG三项检测在血清学诊断鼻咽癌中的效益。方法 收集266例未经治疗的鼻咽癌患者和347例健康成人的血清,用免疫酶法检测VCA-IgA,用酶联免疫吸附法（ELISA）替代免疫酶法检测EA-IgA和EA-IgG抗体。应用新的统计学方法来评估这三项检测的不同优势比。结果 同时应用VCA-IgA,EA-IgG和EA-IgA三项检测的灵敏度和特异度分别为95.11%和97.41%。高于这三项检测的单独应用（VCA-IgA为90.60%和94.52%,EA-IgG为93.98%和93.66,EA-IgA为89.84%和88.18%）,再者,VCA-IgA,EA-IgG和EA-IgA三项检测均呈阳性者的优势比（1912.5)远远高于其中两项检测呈阳性者（VCA-IgA和EA-IgG为27.9032,EA-IgG和EA-IgA为11.1690;VCA-IgA和EA-IgA为8.0328)。而两项检测阳性的优势比又高于单个检测阳性者（VCA-IgA为0.1214;EA-IgG为0.1705;EA-IgA为0.0488)。结论 ELISA法较免疫酶法更能确切反映血清中EB病毒早期抗原的抗体水平。VCA-IgA,EA-IgG和EA-IgA的联合应用能显著提高灵敏度和特异度,明显增高检测阳性的优势比,因此,能有效地提高EB病毒血清学在诊断鼻咽癌时的效益。     

鼻咽癌患者血浆 EBV DNA 水平和 VCA-IgA 检测的临床意义

目的:探讨鼻咽癌患者血浆EBV DNA水平和VCA-IgA联合检测对鼻咽癌早期诊断的临床价值。方法:用荧光定量PCR方法检测血浆EBV DNA水平,常规免疫酶法检测VCA-IgA抗体滴度。结果:68例鼻咽癌患者中,EBV DNA阳性率95.59,中位拷贝数93×104copies/ml,VCA-IgA抗体阳性率92.65,抗体滴度≥1:20;其他头颈肿瘤36例中,EBV DVA阳性率5.56,中位拷贝数为0copies/ml,VCA-IgA抗体阳率36.11,阳性滴度均≤1:20;健康对照组EBV DNA阳性率为35.56,其滴度除3例≥1:40外,余均≤1:20。鼻咽癌患者中EBV DNA和VCA-IgA抗体阳性率显著高于对照组。结论:进一步证实EBV与鼻咽癌有密切关系;EBV DNA水平和VCA-IgA抗体滴度联合检测,有助于鼻咽癌的早期辅助诊断。     

CIK细胞联合白介素-2对鼻咽癌病人免疫功能及EB病毒指标的影响

目的:细胞因子诱导杀伤细胞(cytokine-inducedkillercells,CIK细胞)是目前抗肿瘤过继细胞免疫治疗最为有效的方案。本研究通过治疗30例鼻咽癌病人,探讨CIK细胞联合IL-2对鼻咽癌病人免疫学功能及EB病毒指标的影响。方法:将符合条件的30例病人配对分为两组。治疗组病人接受CIK细胞过继细胞免疫治疗,每周一次,连续4周与对照组比较。免疫功能及EB病毒学指标分别通过测定外周血T淋巴细胞亚群,EB病毒VCA-IgA、EA-IgA、EDAb、DNA免疫荧光定量进行评估。结果:随访8～18个月,中位随访时间12个月。CIK治疗1个月后患者CD4 CD25 %明显下降,EB病毒VCA-IgA、EA-IgA、DNA免疫荧光定量下降显著。结论:CIK细胞可改善鼻咽癌病人的免疫功能,加速EB病毒抗体及DNA拷贝数下降。提示CIK治疗可消灭肿瘤微小残留灶及转移灶,可作为一种鼻咽癌的辅助治疗。     

检测EB病毒VCA-IgA抗体在提高鼻咽癌检出率上的意义

在鼻咽癌患者血清中,常可测到高滴度的抗Epstein-Barr(EB)病毒多种特异性抗原的抗体。其中,抗EB病毒壳抗原(VCA)的特异性免疫球蛋白A抗体VCA-IgA更具特异性,已被广泛应用于临床。鼻咽癌患者血清VCA-IgA抗体的阳性率达90.14％及95.07％。然而,这一血清学诊断不能单独作为确诊鼻咽癌的依据,只是一种辅助方法。为了确定其在帮助发现鼻咽癌病例上的意义,我们     

EBV-DNA检测与鼻咽癌咽后淋巴结及颈部淋巴结转移的关系

[目的]探讨EB病毒与鼻咽癌淋巴结转移的关系。[方法]收集首诊鼻咽癌患者721例,按"中国鼻咽癌2008TNM分期"标准对咽后淋巴结及颈部淋巴结转移进行判断,分别计算咽后淋巴结阴阳性与颈部淋巴结阴阳性的病例数;使用PCR检测法对病例进行治疗前血浆EBV-DNA检测,以及记录每个病例EBV-DNA拷贝数。[结果]鼻咽癌患者咽后淋巴结转移者有较高的颈部淋巴结转移率,差异有统计学意义(P=0.013),Pearson关联系数,r=0.342,P=0.000,有统计学意义;双侧咽后淋巴结转移较单侧转移者有较高的颈部淋巴结转移率,差异有显著统计学意义(P=0.001);咽后淋巴结与颈部淋巴结均有转移者较其他病例组有较高的EBV-DNA拷贝数,差异有显著统计学意义(P=0.000)。[结论]鼻咽癌患者咽后淋巴结转移与颈淋巴结转移有正相关关系,咽后淋巴结与颈部淋巴结均有转移者有较高的EBV-DNA拷贝数。推测EBV-DNA检测可作为评估鼻咽癌淋巴结转移的分子生物学指标之一。     

EB病毒血清学检测对鼻咽癌的诊断价值

自1966年 Old 等首先发现鼻咽癌与 EB 病毒在血清学上有关以来,EB 病毒与鼻咽癌的密切关系即被充分肯定。鼻咽癌病人血清中多种EB 病毒特异的抗体滴度都升高,特别是 EB 病毒壳抗原的免疫球蛋白 A(VCA-IgA)抗体具有较高的特异性,对鼻咽癌的早期诊断有一定的价值。本文仅就近年来的研究概况加以综述。Henle 等首先报告 VCA-IgA 抗体对鼻     

鼻咽癌患者血浆EBV DNA、血清CYFRA21-1和VCA-IgA的检测及临床应用

目的：探讨血浆EB病毒DNA、血清细胞角蛋白19片段（CYFRA21-1）和EB病毒壳抗原IgA（VCA-IgA）三种肿瘤标记物对鼻咽癌诊断、疗效监测和预后判断的临床应用价值。方法：分别采用荧光定量PCR法、电化学发光法和免疫酶法检测62例鼻咽癌患者治疗前、后及62例健康对照者血浆EBV DNA、血清CY-FRA21-1和VCA-IgA含量,并进行对比分析。结果：治疗前血浆EBV DNA、血清CYFRA21-1和VCA-IgA的阳性率明显高于健康对照组（P〈0.01）;治疗后血浆EBV DNA、血清CYFRA21-1和VCA-IgA含量高者,在随访中发现肿瘤残存、复发或远处转移。结论：血浆EBV DNA、血清CYFRA21-1和VCA-IgA的检测对鼻咽癌早期诊断、高危人群筛查、以疗效监测和预后判断有重要临床应用价值,三种肿瘤标记物联合检测有互补作用。     

Serologic diagnosis of nasopharyngeal carcinoma. A double-blind study of four EB virus antibodies with evaluation by sequential discrimination

It is generally known that a close relationship exists between Epstein-Barr Virus (EBV) and nasopharyngeal carcinoma (NPC). Recently, patients with early lesions of NPC have been detected in the general population by use of serologic mass survey. Using the double-blind method, we have studied the diagnostic value of the four EBV antibody titers, VCA-IgA, VCA-IgG, EA-IgA and EA-IgG, in four groups of subjects, each consisting of 50 persons: patients with nasopharyngeal carcinoma (NPC group), patients with cancers other than NPC in the head and neck regions (HNC group), patients with cancers outside of head and neck regions (OC group) and normal individuals (NS group). The results of these four antibodies were evaluated both singularly and together by multivariate sequential discrimination. Taking 1:10 as the criterion of being positive, in the NPC group, the positive rate of VCA-IgA is 88%, the VCA-IgG rate is 100%, the EA-IgA rate is 48% and the EA-IgG rate is 74%. In the non-NPC group, the positive rates of VCA-IgA are as high as 86%-92%, but those of the other antibodies are as low as 0-42%. The positive rates and the geometric mean titers of these four antibodies were all elevated as compared with those in the three non-NPC groups. These differences are statistically significant. VCA-IgG is unimportant in the diagnosis of NPC because of its low specificity. By treating the antibody titers of VCA-IgA, VCA-IgG, EA-IgA and EA-IgG with sequential discrimination, the correlation rate between the serology and pathology of NPC is 88% and the false positive rate is 7.3%.     

Gene engineering EB virus membrane antigen in detection of MA-IgA antibody

With gene engineering EB virus membrane antigen as the diagnostic antigen, indirect immunofluorescence (IF) assay was used to detect IgA antibody against EB virus membrane antigen (MA-IgA) in sera from 202 nasopharyngeal carcinoma (NPC) patients and 315 controls (normal and patients with other tumors). MA-IgA antibody was positive in 96.8% of the pretreatment NPC patients with a GMT of 1:36.3. MA-IgA detection by this method was more sensitive than EA-IgA detection by IE. In contrast, patients with tumors other than NPC were negative for MA-IgA antibody. 9.1% of VCA-IgA positive persons were MA-IgA positive with a GMT of less than 1:5. No MA-IgA positive was found in VCA-IgA negatives. The results indicated that this method was relatively specific. In the treatment group, the positive rate and GMT of MA-IgA antibody declined with increase in survival time and the decline was faster than VCA-IgA. When recurrence or distant metastasis developed, similar to VCA-IgA and EA-IgA antibodies, the positive rate and GMT of MA-IgA antibody increased to its pretreatment level. Therefore, MA-IgA detection might be valuable in the early diagnosis and monitor of NPC.     

Establishment of VCA and EBNA1 IgA-based combination by enzyme-linked immunosorbent assay as preferred screening method for nasopharyngeal carcinoma: a two-stage design with a preliminary performance study and a mass screening in southern China

A two-stage study was conducted in southern China to determine and validate an optimal combination of Epstein-Barr virus (EBV)-related seromarkers for nasopharyngeal carcinoma (NPC) screening. In the first stage, six seromarkers [VCA-IgA, EA-IgA, Epstein-Barr virus nuclear antigen 1 (EBNA1-IgA), EBNA1-IgG, Zta-IgA and Rta-IgG] were detected by enzyme-linked immunosorbent assay (ELISA) and two traditional NPC screening seromarkers (VCA-IgA and EA-IgA) were detected by immunofluorescence assay (IFA) in serum samples from 191 NPC patients and 337 controls. An optimal combination of seromarkers for NPC diagnosis was selected using logistic regression models. Results showed that the diagnostic performances of VCA-IgA and EA-IgA tested by ELISA were superior to the performances of the same seromarkers by IFA. VCA-IgA combined with EBNA1-IgA by ELISA was identified as the optimal combination, with an area under the receiver operating characteristic (ROC) curve (AUC) up to 0.97, a sensitivity of 95.3% and a specificity of 94.1% for classification of NPCs vs. controls. In the second stage, 5,481 participants aged 30-59 years and without clinical evidence of NPC were recruited into a population-based NPC screening program from May 2008 to February 2009 in Sihui City, China. Their sera were tested simultaneously by both the new and the traditional screening schemes and eight early stage NPC patients were subsequently histopathologically confirmed. The traditional and the new screening schemes had comparable specificity (estimated as 98.5%), but the sensitivity of the new scheme (75.0%) was significantly higher than that of the traditional one (25.0%). The combination of VCA-IgA and EBNA1-IgA by ELISA outperforms the traditional NPC screening scheme and could become the preferred serodiagnostic strategy for NPC screening in high-incidence areas.     

基因工程膜抗原检测EB病毒MA—IgA抗体研究及与VCA—IgA...

With gene engineering EB virus membrane antigen as the diagnostic antigen, indirect immunofluorescence (IF) assay was used to detect IgA antibody against EB virus membrane antigen (MA-IgA) in sera from 202 nasopharyngeal carcinoma (NPC) patients and 315 controls (normal and patients with other tumors). MA-IgA antibody was positive in 96.8% of the pre-treatment NPC patients with a GMT of 1:36.3. MA-IgA detection by this method was more sensitive than EA-IgA detection by IE. In contrast, patients with tumors other than NPC were negative for MA-IgA antibody. 9.1% of VCA-IgA positive persons were MA-IgA positive with a GMT of less than 1:5. No MA-IgA positive was found in VCA-IgA negatives. The results indicated that this method was relatively specific. In the treatment group, the positive rate and GMT of MA-IgA antibody declined with increase in survival time and the decline was faster than VCA-IgA. When recurrence or distant metastasis developed, similar to VCA-IgA and EA-IgA antibodies, the positive rate and GMT of MA-IgA antibody increased to its pretreatment level. Therefore, MA-IgA detection might be valuable in the early diagnosis and monitor of NPC.     

The association between circulating tumor cells and Epstein-Barr virus activation in patients with nasopharyngeal carcinoma

Background: Circulating tumor cells (CTCs) and microemboli (CTM) are attracting increasing attention in medical biology and clinical practice. However, the clinical relevance of CTCs in nasopharyngeal carcinoma (NPC) has not yet been ascertained, and no study has focused on the influence of Epstein-Barr virus (EBV) status on CTCs in NPC patients. These issues were therefore examined. Methods: Peripheral blood samples were prospectively obtained from 33 NPC patients before treatment. CTCs and CTM were captured using the Isolation by Size of Epithelial Tumor (ISET) method. Immunohistochemistry on CK5/6 (cytokeratin5/6) and P63, as well as in situ hybridization of EBERs (EBV-encoded RNAs) were used to validate the harvested tumor cells. Results: Out of 33 NPC patients, CTCs were detected in 22 cases (66.7%), and CTM were observed in 2 cases (6.1%). CTCs were presented in all stages of NPC patients but had no association with the TNM stages (all P > 0.05). The presence of CTCs appeared to correlate with EBV activation status. Among the total NPC patients, the EBV VCA-IgA levels in CTC-positive cases were higher than that in CTC-negative cases (negative vs. positive: 3.88 vs. 4.86, P = 0.016). A similar result was observed in the patients without distant metastasis (negative vs. positive: 3.76 vs. 4.95, P = 0.009). When the number of CTCs was considered, CTC counts were found to correlate with EBV VCA-IgA (R = 0.382, P = 0.041) and EBV DNA load (R = 0.396, P = 0.033) for all NPC patients as well as NPC patients without distant metastases. Conclusions: These findings strongly suggested detectable CTCs/CTM in all stages of NPC patients and support a positive correlation between CTCs and EBV activation in NPC patients.