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。     

EA-D p45-IgG as a Potential Biomarker for Nasopharyngeal Carcinoma Diagnosis

Aim: To identify new biomarkers for NPC diagnosis with an anti-EBV Western blot test kit. Methods:Serum samples from 64 NPC patients and healthy subjects with four specific VCA-IgA/EA-IgA profiles weretested with an anti-EBV Western blot test kit from EUROIMMUN AG. Proteins were quantified with scores ofintensity visually assigned to the protein bands. The markers which showed statistical differences between theNPC and non-NPC subjects were further evaluated in another 32 NPC patients and 32 controls in comparisonwith established biomarkers including VCA-IgA, EA-IgA, EBV-related protein IgG, and EBV DNA. Results:Among the markers screened, EA-D p45-IgG showed a statistically significant difference (p < 0.05) between NPCand non-NPC subjects with VCA-IgA positivy. In 32 VCA-IgA positive NPC patients and 32 control subjects,the diagnostic accuracy of EA-D p45-IgG was 78.1% with a positive predictive value of 77.8% and a negativepredictive value of 78.6%. In the verification experiment, the specificity and sensitivity of EA-D p45-IgG were75.0% and 90.6 %, respectively. Conclusions: EA-D p45-IgG might be a potential biomarker for NPC diagnosis,especially among VCA-IgA positive subjects.     

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.     

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.     

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。     

Comparison of the prognostic impact of serum anti-EBV antibody and plasma EBV DNA assays in nasopharyngeal carcinoma

PURPOSE: Nasopharyngeal carcinoma (NPC) has been proven as an Epstein-Barr virus (EBV)-associated cancer. Serum anti-EBV antibodies and plasma EBV DNA have been investigated as surrogate markers for NPC. A comparison of the prognostic impacts of both assays has never been reported. METHODS AND MATERIALS: Paired serum and plasma samples from 114 previously untreated NPC patients were collected and subjected to an immunofluorescence assay for immunoglobulin (Ig)A and IgG antibodies against the viral capsid antigen (VCA) and a real-time quantitative polymerase chain reaction assay for EBV DNA measurement. The effects of both assays on patient prognosis were thoroughly investigated. RESULTS: Relapsed patients had significantly higher pretreatment EBV DNA concentration than patients without relapse (p = 0.0006). No associations of VCA-IgA (p = 0.9669) or VCA-IgG (p = 0.6125) were observed between patients with and without relapse. The 4-year overall survival (60.3% vs. 93.1%, p < 0.0001) and relapse-free survival rates (54.4% vs. 77.9%, p = 0.0009) were significantly lower in patients with higher pretreatment EBV DNA load than in those with lower EBV DNA load. Patients with persistently detectable EBV DNA after treatment had significantly worse 4-year overall (30.8% vs. 84.6%, p < 0.0001) and relapse-free survival rates (15.4% vs. 74.0%, p < 0.0001) than those with undetectable EBV DNA. The VCA-IgA and VCA-IgG titer could not predict survivals (all p > 0.1). Cox multivariate analyses also showed the same results. CONCLUSION: Plasma EBV DNA is superior to serum EBV VCA antibodies in prognostic predictions for NPC.     

鼻咽癌咽后淋巴结转移与 EBV-DNA 血清学标记物关系探讨

目的:探讨鼻咽癌患者咽后淋巴结转移与EB病毒血清学标志物的关系.方法:收集首诊鼻咽癌患者721例,按“中国鼻咽癌2008 TNM分期”标准对咽后淋巴结进行判断并分为咽后淋巴结转移阳性组与阴性组,分别记录每个病例EB病毒血清学标记物:EB-DNA拷贝数、EB-VCA抗体滴度、EA-IgA抗体滴度,统计咽后淋巴结有转移组与无转移组与EB病毒血清学标记物:EB-DNA拷贝数、VCA-IgA抗体滴度、EA-IgA抗体滴度的关系.结果:鼻咽癌患者咽后淋巴结阳性者较阴性组有较高的EB-DNA拷贝数,差异有显著统计学意义(P＜0.01);不同VCA-IgA抗体滴度下咽后淋巴结的阳性率不同,差异有统计学意义(P＜0.05),VCA-IgA抗体滴度越高咽后淋巴结阳性率越高;不同EA-IgA抗体滴度下咽后淋巴结的转移率不同,差异有显著统计学意义(P＜0.05),EA-IgA抗体滴度越高咽后淋巴结阳性率越高.结论:鼻咽癌患者咽后淋巴结转移与EB病毒血清学标记物:EB-DNA拷贝数、VCA-IgA抗体滴度、EA-IgA滴度呈正相关.     

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.     

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。     

EB病毒感染与早期鼻型结外NK/T细胞淋巴瘤患者疗效及预后的关系

目的 探讨早期结外鼻型NK/T细胞淋巴瘤（ENKTCL）患者治疗前EBV DNA载量、治疗前血清EA-IgA及VCA-IgA抗体水平与临床特征、治疗反应及预后的关系。方法 分析78例早期结外鼻型NK/T细胞淋巴瘤患者的临床特征及影响预后的因素。结果 治疗前EBV DNA、VCA-IgA、EA-IgA阳性率分别为43.6%、20.5%、14.1%。EBV DNA与Ann Arbor分期、原发部位、PTI、治疗后未获得CR显著相关（均P<0.05）。VCA-IgA、EA-IgA滴度分别与EBV DNA、治疗后未获得CR显著相关（均P<0.05）。多因素分析发现年龄、EBV DNA、治疗后未获得CR为早期ENKTCL患者OS的独立预后因素（均P<0.05）;年龄、EBV DNA、原发鼻腔外上呼吸消化道、治疗后未获得CR（均P<0.05）则为早期ENKTCL患者PFS的独立预后因素。结论 治疗前EBV DNA阳性与较晚的Ann Arbor分期、PTI、原发鼻腔外上呼吸消化道、治疗反应差有关。EA-IgA、VCA-IgA水平升高与EBVDNA阳性、治疗反应差有关。治疗前EBV DNA可用于ENKTCL的风险分层及预后预测,而EA-IgA、VCA-IgA对于ENKTCL的预后指导作用有限。     

Immunity to antigens associated with a cell line derived from nasopharyngeal cancer (NPC) in non-Chinese NPC patients.

Delayed hypersensibility to antigens derived from four lymphoid cell lines was measured in 27 non-Chinese patients with nasopharyngeal cancer (NPC) and 63 non-NPC cancer patients. Of the NPC patients, 17/27 (63%) had a positive skin test response to antigens derived from HKLY-28, a lymphoid cell line which was developed from an NPC biopsy. Only 10/51 (20%) and 1/13 (8%) patients with solid tumors and hematopoietic malignancies, respectively, had positive skin test responses to HKLY-28. Positive skin tests were found less frequently when extracts from cell lines derived from normal individuals or lymphoma patients were utilized, although NPC patients were more reactive to two of the non-NPC derived cell lines than the controls. The NPC patients in this study also had significantly elevated antibody titers to the Epstein-Barr virus (EBV), capsid antigen (VCA) and early antigen (EA). Titers were highest in the patients with more anaplastic nasopharyngeal carcinomas. The skin test and serologic data are consistent with studies in Chinese patients, indicating that NPC in non-Chinese and Chinese patients is biologically similar.     

A Follow－up Survey of 42048 Subjects Tested with Epstein－Barr Viral Serology in a High －incidence Area of Nasopharyngeal Carcinoma

Objective:To evaluate the efficiency of Epstein-Bar viral serology in detecting nasopharyngeal carcinoma(NPC)and comparing the NPC risks between serology-positive and serology-negative subjects.MethidsLUnnybiebztnatuc tutratuib for VCA-IgA was performed in 42048 subjects between the ages of 30 and 59.The titre of VCA-IgA,≥1/5,EA-IgA,≥1/5 was recognized as being positive.All those were followed up to the end of 1999.Results:45 NPCs were detected in the first year(45/42048,107.02/10^5).The average annual incidence of NPC in 3050 VCA-IgA-positive subjects(144.86/10^5)was higher than that(11.98/10^5)in 38953 VCA-IgA-negative subjects during the next 12 years.72 out the 154 NPC patients detected were of stage-I.Conclusion :This reveals that EBV serology could actually detect early-stage NPCs as previously reported.Meaningfully,the NPC risk increased 12 times the VCA-IgA-negative subjects in the VCA-IgA-positive subjects after the first-turn serological screening.The 5-year and 10-year survival rates of stage-I NPC patients(81.25%,67.39% respectively)were both significantly higher than those of the stage-II A, stage-II B and stage-Ⅲ patients (60.32%,26.67% respectively)after radiotherapy.     

Antibodies to the Epstein-Barr virus transactivator protein (ZEBRA) as a valuable biomarker in young patients with nasopharyngeal carcinoma

Epstein-Barr virus (EBV)-associated nasopharyngeal carcinoma (NPC) generally occurs in adults, especially in high-prevalence populations such as the Chinese and Eskimos. In Maghrebian populations, young patients affected with this malignancy represent 25% of the total NPC cases. In adults with NPC, relatively high titers of IgA antibodies to the EBV viral capsid antigen (VCA) and early antigen (EA) represent important markers. However, nearly 50% of young NPC patients are negative for IgA-anti-VCA and -EA or exhibit very low titers of these antibodies. We report here that 92% of sera from young NPC patients negative for IgA-EA and 89% of those negative for IgA-VCA were positive for IgG antibodies to the EBV transactivator protein (ZEBRA) at very high titers. Our results show that in young patients with NPC these antibodies represent the most reliable marker for diagnosis and prognosis, particularly when compared with conventional NPC markers, i.e., IgA-VCA (58%) and anti-EA (25%). The titers of IgG-ZEBRA antibodies increased along with lymph node involvement only in the young patient group, suggesting a prognostic value of this marker in this patient group.     

Titers of Epstein-Barr virus-related antibodies in nasopharyngeal carcinoma in Japan

It is thought that nonkeratinizing or undifferentiated squamous cell carcinoma in the nasopharynx (NPC) is intimately correlated with Epstein-Barr Virus (EBV). Twenty-one patients with NPC were followed in Kyoto University Hospital and 4 in Osaka Red Cross Hospital during the past 2 years from 1980 to 1981. These patients were classified histopathologically according to the WHO classification in 1978 and staged with the TNM classification in Union Internationale Contre le Cancer (UICC) in 1978. The incidence rate of NPC among the head and neck tumors was 5.6% in the authors' university from 1980 to 1981. The sex ratio of male to female was nearly equal. The mean age of NPC patients was 56.7 years. Sera from these 25 patients with nasopharyngeal carcinoma were collected at intervals of 3 to 8 months over a 2-year period, and were examined for their spectra and titers of antibodies of EBV-related antigens. They were titrated for IgG, IgA and IgM antibodies to EB viral capsid antigen (VCA), for IgG and IgA antibodies to early antigen-DR component (EA) and for antibodies to EBV-associated nuclear antigen (EBNA). All of these patients were primarily treated with radiation, while a few who did not respond to this therapy were subsequently treated with surgery or chemotherapy. EBV antibodies of VCA-IgG, -IgA, EA(DR)-IgG, and -IgA and EBNA were elevated in 73% and 90% of the nonkeratinizing and undifferentiated NPC patients, respectively. The VCA-IgM was elevated in almost none of the cases. In contrast to this, these values were all in a normal range in the NPC patients with keratinizing squamous cell carcinoma and malignant lymphoma. Also 9% and 10% of nonkeratinizing and undifferentiated carcinomas showed the normal ranges of EBV antibodies, possibly indicating a nonassociation with EBV. When NPC disappeared with radiation therapy, EBV antibodies became normal for 6-18 months. However, those whose NPC did not respond to the combined therapy with radiation, surgery and chemotherapy maintained high titers of EBV antibodies. The prognosis was the poorest in the patients with undifferentiated carcinoma, 40% of whom died within 4 years after diagnosis.     

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。     

89Sr和153Sm-EDTMP治疗鼻咽癌多发性骨转移瘤疗效的比较

目的 比较＾８９Ｓｒ和＾１５３Ｓｍ－ＥＤＴＭＰ治疗鼻咽癌（ｎａｓｏｐｈａｒｙｎｇｅａｌ ｃａｒｃｉｎｏｍａ,ＮＰＣ）多发性骨转移瘤的疗效。方法 从１９９７年５月到１９９９年３月,作者用＾８９Ｓｒ和＾１５３Ｓｍ－ＥＤＴＭＰ分别治疗ＮＰＣ多发性骨转移病人４５例和６８例。根据治疗前后骨痛缓解和生活质量的情况决定疗效,同时比较其病灶摄取放射性变化,观察开始起效的时间,对病人血清ＶＣＡ－ＩｇＡ、ＥＡ－ＩｇＡ     

Comparison of plasma Epstein-Barr virus (EBV) DNA levels and serum EBV immunoglobulin A/virus capsid antigen antibody titers in patients with nasopharyngeal carcinoma

BACKGROUND: Serologic measurement of antibodies to Epstein-Barr virus (EBV) immunoglobulin A/viral capsid antigen (IgA/VCA) and early antigen (IgA/EA) has been used widely to screen for nasopharyngeal carcinoma (NPC) in China. Recently, it was found that plasma EBV DNA concentration is an indicator for the staging and prognosis of patients with NPC. To determine whether there is a correlation between plasma EBV DNA levels and serum levels of IgA/VCA, the authors measured both in patients with NPC and in a control group. METHODS: Real-time polymerase chain reaction was used for quantitative analysis of plasma EBV DNA concentration, and enzyme-linked immunoadsorbent assay was used to measure EBV VCA/IgA in patients with primary NPC (n = 120 patients), locally recurrent NPC (n = 8 patients), and distant metastatic NPC (n = 21 patients) among 76 patients with NPC after the completion of radiotherapy, in 60 patients with NPC in clinical remission, in 38 patients with non-NPC tumors, and in 47 control individuals. RESULTS: The median plasma EBV DNA levels were 6200 copies/mL, 9200 copies/mL, and 2050 copies/mL in patients with primary, locally recurrent, and distant metastatic NPC, respectively, but declined to 0 copies/mL in patients with clinically remissive NPC, in patients who completed radiotherapy, in patients with non-NPC tumors, and in the control group. In contrast, EBV VCA/IgA titers and detection rates remained high in all NPC groups. Plasma EBV DNA levels were significantly higher in patients who had serum VCA/IgA titers > or = 1:640 (median, 83,450 copies/mL) compared with the levels in patients who had titers < or = 1:320 (median, 17,200 copies/mL). Patients with NPC who had advanced TNM stage (Stages III and IV; median, 8530 copies/mL) and T classification (T3 and T4 tumors; median, 8530 copies/mL) had significantly higher plasma EBV DNA levels compared with patients who had early TNM stage (Stages I and II; median, 930 copies/mL) and T classification (T1 and T2 tumors; median, 3700 copies). Patients who had advanced TNM stage NPC had significantly higher mean VCA/IgA titers (1:424) compared with patients who had early TNM stage NPC (1:246), but there was no correlation between IgA/VCA titer and T or N classification of NPC. CONCLUSIONS: The results suggest that plasma EBV DNA detection is a more sensitive and specific marker than the serum IgA/VCA titer for the diagnosis and monitoring of patients with NPC. These findings provide convincing evidence for the use of plasma EBV DNA measurements for the early diagnosis and staging of NPC as well as for monitoring recurrence and metastasis of this tumor.     

抗EB病毒VCA—IgA抗体检测结果一致性检查

本文报告广东省五个鼻咽癌普查现场组,以各自检测抗EB病毒VCA—IgA抗体的试剂盒,应用免疫酶法,同时检测中山市普查的180份正常人血清,各现场组的检测结果和中山市提供的血清的滴度,通过一致率和Kappa值计算,一致率(P0)为66—86％,Kappa值(K)为0.28—0.48,U为3.14—6.28,P<0.01。结果表明所得的K值均能排除因机遇所致的可能,其一致性是有意义的,从而提示各现场检测的抗EB病毒VCA—IgA抗体的阳性率可提供现场间的比较。 本文还对检测结果和可能影响VCA—IgA抗体检测一致性的因素进行了讨论。     

Noninvasive diagnosis of nasopharyngeal carcinoma: nasopharyngeal brushings reveal high Epstein-Barr virus DNA load and carcinoma-specific viral BARF1 mRNA

Nasopharyngeal carcinoma (NPC) is the most prevalent ENT-tumour in Indonesia. We investigated the primary diagnostic value of Epstein-Barr virus (EBV) DNA load and mRNA detection in noninvasive nasopharyngeal (NP) brushings, obtained prospectively from consecutive Indonesian ENT-patients with suspected NPC (N=106) and controls. A subsequent routine NP biopsy was taken for pathological examination and EBER-RISH, yielding 85 confirmed NPC and 21 non-NPC tumour patients. EBV DNA and human DNA load were quantified by real-time PCR. NP brushings from NPC patients contained extremely high EBV DNA loads compared to the 88 non-NPC controls (p<0.0001). Using mean EBV DNA load in controls plus 3 SD as cut-off value, specificity, sensitivity, positive and negative predictive values were 98, 90, 97 and 91%, respectively. Epstein-Barr nuclear antigen 1 (EBNA1) and the carcinoma-specific BARF1 mRNA were detected by nucleic acid sequence based amplification and found in 86 and 74% of NP brushings, confirming NPC tumour cell presence. EBV RNA positivity was even higher in fresh samples stored at -80 degrees C until RNA expression analyses (88% for both EBNA1 and BARF1). EBV RNA-negative NP brushings from proven NPC cases had the lowest EBV DNA loads, indicating erroneous sampling. No EBV mRNA was detected in NP brushings from healthy donors and non-NPC patients. In conclusion, EBV DNA load measurement combined with detection of BARF1 mRNA in simple NP brushings allows noninvasive NPC diagnosis. It reflects carcinoma-specific EBV involvement at the anatomical site of tumour development and reduces the need for invasive biopsies. This procedure may be useful for confirmatory diagnosis in large serological NPC screening programs and has potential as prognostic tool.