血清EB病毒抗体水平与鼻咽癌患者预后的关系

目的 探讨血清EB病毒VCA/IgA、EA/IgA、NA1/IgA及Rta/IgG抗体水平与鼻咽癌患者预后的关系.方法 140例初治无远处转移的鼻咽癌患者分别在治疗前和治疗结束后采用免疫酶法检测血清VCA/IgA和EA/IgA,ELISA法检测NA1/IgA和Rta/IgG.随访进行远期疗效和生存的评价.结果 治疗后患者血清VCA/IgA、EA/IgA、NA1/IgA及Rta/IgG抗体水平较治疗前有明显下降,但仍显著高于正常对照组(P＜0.05).治疗后持续缓解的鼻咽癌患者其治疗前VCA/IgA、EA/IgA抗体水平显著低于疾病进展患者(P＜0.05).血清VCA/IgA、EA/IgA、NA1/IgA及Rta/IgG抗体水平与患者的3年总生存率无关(P＞0.05).治疗前VCA/IgA抗体高水平组(≥1∶320)及EA/IgA抗体高水平组(≥1∶80)患者的无进展生存期(61.8％,61.3％)低于抗体低水平组患者(86.5％,86.5％;P＜0.001).Cox回归分析显示治疗前VCA/IgA抗体水平是影响无进展生存的独立危险因素(HR=3.80,P=0.001).结论 VCA/IgA、EA/IgA可为鼻咽癌患者预后判断提供帮助.     

鼻咽癌EB病毒Rta/IgG抗体检测的诊断价值

目的 通过检测血清中的EB病毒Rta/IgG抗体,评价其在鼻咽癌诊断上的价值.方法 收集211例未经治疗的鼻咽癌患者,413例对照组(包括203例相似症状的非鼻咽癌病例和210例健康体检者)的血清,用酶联免疫吸附法(ELISA)检测Rta/IgG抗体.应用受试者工作特征(ROC)曲线对结果进行分析评价.结果 鼻咽癌组的Rta/IgG抗体rA值中位数明显高于对照组(P<0.001).Rta/IgG抗体检测诊断鼻咽癌的ROC曲线下面积为0.933,最佳截断点时敏感度为90.5%,特异度为90.1%.结论 采用ELISA方法检测血清中Rta/IgG抗体可以作为检测EB病毒的一个新指标,并可作为鼻咽癌诊断的重要标志物之一.     

五厂家EB病毒抗体检测试剂盒血清学诊断鼻咽癌的比较

目的 通过比较EB病毒抗体检测试剂盒血清学诊断鼻咽癌的准确性和检测结果的一致性,为试剂盒在临床上的使用选择和性能改进提供依据.方法 使用五厂家的EB病毒衣壳抗原IgA和IgG抗体检测试剂盒（VCA IgA和VCA IgG试剂盒）、核抗原I IgA和IgG抗体检测试剂盒（EBNA1 IgA和EBNA1IgG试剂盒）、早期抗原IgA和IgG抗体检测试剂盒（EA IgA和EA IgG试剂盒）以及Zta IgA抗体检测试剂盒,分别检测33例鼻咽癌患者（NPC）、30例健康体检者（HD）和41例非鼻咽癌的其他肿瘤患者（NNPC）血清或血浆样本.结果 A厂家的VCA IgA试剂盒灵敏度高于其他厂家同品种试剂盒,但对于NNPC特异度最低（36.6％）;而D厂家VCA IgA试剂盒的特异度最高（97.6％）,且对HD的特异度均大于90％.B和D厂家的EBNA1 IgA试剂盒间阳性、阴性符合率分别为92.1％和100.0％.A和E厂家的EA IgA试剂盒的灵敏度均较低而特异度高,试剂盒间阳性符合率低（39.4％）,阴性符合率高（98.6％）;而VCA IgG试剂盒的灵敏度高但特异度低.A和C厂家的EBNA1IgG试剂盒的灵敏度高（100.0％,97.0％）但特异度低（3.3％,13.3％）.C厂家EA IgG试剂盒检测所有样本结果均为阴性.结论 五个不同厂家VCA LgA、EA IgA试剂盒诊断鼻咽癌的准确性和检测结果的一致性存在差异,特别是A厂家和其他国内厂家同品种试剂间差异明显,需根据临床目的进行选择.三家国产VCA IgA试剂盒的灵敏度需进一步提高.相反,EBNA IgA试剂盒诊断鼻咽癌的准确性和结果一致性较好.单独使用VCA IgG和EBNA1 IgG试剂盒血清学诊断鼻咽癌的特异度差,其判读界值可能需根据检测且的进行调整.     

基于2008分期标准的壮族鼻咽癌各期患者中EB病毒Zta/IgG等抗体阳性率的比较

探讨桂西地区壮族鼻咽癌患者EB病毒EBNA1/IgA、Zta/IgG及Rta/IgG抗体阳性率与鼻咽癌2008分期的关系。收集140例未经治疗的鼻咽癌患者的血清,用酶联免疫吸附法(ELISA)检测EBNA1/IgA、Zta/IgG及Rta/IgG抗体,按"2008分期法"进行分期,分别计算临床分期的各抗体阳性率并进行两两对比,进行统计学分析。鼻咽癌各临床分期组EB-NA1/IgA、Zta/IgG及Rta/IgG抗体阳性率均有统计学差异(P<0.05)。III期的各抗体阳性率明显低于IV期临床分期(P<0.05),Rta/IgG抗体有显著差异(P=0.001)。壮族鼻咽癌患者EB病毒EBNA1/IgA、Zta/IgG及Rta/IgG抗体阳性率与鼻咽癌临床分期有关。或许EBNA1/IgA、Zta/IgG及Rta/IgG抗体对于评估鼻咽癌临床分期有一定的参考价值。     

鼻咽癌血清学检测方法的改进

目的 改进现有鼻咽癌血清学早期诊断方法,提高检测灵敏度.方法 用链霉亲和素.生物素放大免疫酶法与既有常规免疫酶法对鼻咽癌防治示范基地所取294份普查血清IgA/VCA,IgA/EA抗体进行血清学检测,并用SPSS统计软件对检测结果进行χ2检验和t检验.结果 共检测30岁以上294份普查人群血清,其中鼻咽癌患者血清106份,健康人血清188份.改进的链霉亲和素一生物素放大免疫酶法与既往已有的免疫酶法相比,血清IgA/VCA、IgA/EA抗体的检测阳性率增加;检测血清抗体的几何平均数明显提高.结论 改进方法可以提高血清学检测的灵敏度,同时保证了检测结果的特异性,提高了鼻咽癌的检出率,可以应用于鼻咽癌早期筛查工作.     

EBV4型IgA/VCA IgA/EA IgG/EA IgG/ZEBRA抗体在鼻咽癌普查和早期诊断中的应用

目的摸索以疱疹病毒4型（EBV）IgG／ZEBRA为捕捉抗原的间接酶联免疫吸附试验（ELISA）条件,为大量人群普查奠定基础。方法将纯化的ZEBRA抗原用于对鼻咽癌（NPC）患者血清及健康人血清IgG／ZEBRA抗体的ELISA检测。结果检测NPC患者血清288份,其中ELISA实验显示阳性262份,敏感度91％,检测正常人血清96份,其中阳性5份,特异度94．8％。其结果显示NPC组的阳性率与健康对照组的数据之间差异有统计学意义（P〈0．001）。本研究在此基础上对广东惠州5463份和广西桂平2017份血清进行检测,检出早期鼻咽癌患者5例。并将结果与免疫酶法检测IgA／VCA、IgA/EA、IgG／EA比较。结论以EBV早期抗原ZEBRA为捕捉抗原的间接ELISA方法具有较高的特异性和敏感性,可以用于大量人群的NPC早期筛查和早期诊断。     

Logistic回归结合ROC曲线在HBV相关肝硬化合并肝癌患者中的诊断价值

研究以治疗前170例HBV相关肝硬化合并肝癌、90例HBV相关肝硬化和90例慢性乙型肝炎(chronic hepatitis B, CHB)患者为研究对象,检测异常凝血酶原(abnormal prothrombin,PIVKA-Ⅱ)、高尔基体蛋白73(Golgi protein 73,GP73)、AFP和甲胎蛋白异质体(alpha fetoprotein heterogeneity,AFP-L3)的血清水平,建立单项检测的ROC曲线,确定各指标的最佳诊断临界值(cutoff值),再通过Logistic回归评价多变量检测模型的诊断价值。结果表明,单项检测时,PIVKA-Ⅱ的ROC曲线下面积(area under the curve, AUC)最大(0.920),敏感度和特异度分别为91.2%和85.6%。多项检测时,PIVKA-Ⅱ联合AFP的Logistic回归方程为最佳诊断模型,ROC AUC最大(0.951),敏感度和特异度分别为87.6%和91.1%,优于两项指标的并联和串联实验。肝癌风险概率值P=1/[1+e~(-(-5.284+9.087×PIVKA-Ⅱ+4.756×AFP))],在早期(Ⅰ期和BCLC A期)肝癌患者中的敏感度为66.7%,在小肝癌(肿瘤直径3 cm)患者中的敏感度为67.7%。因此,PIVKA-Ⅱ联合AFP的Logistic回归方程诊断模型在HBV相关肝硬化合并肝癌患者中具有良好的诊断价值,利用肝癌风险概率值可从良性肝病患者中及时诊断出早期肝癌和小肝癌。     

应用Logistic回归和ROC曲线综合分析PSA、fPSA、fPSA/PSA和EPCA对前列腺癌的诊断价值

目的 探讨Logistic回归和ROC曲线综合分析PSA、fPSA、fPSA/PSA和EPCA对前列腺癌的诊断价值.方法 检测107例前列腺癌(prostate cancer,PCa)患者、84例良性前列腺增生(benign prostatic hyperplasia,BPH)患者和50例健康对照者的血清PSA、fPSA和EPCA水平,计算fPSA/PSA,建立Logistic回归模型,绘制ROC曲线并计算曲线下面积来评价各肿瘤标志物对PCa的诊断价值.结果 PCa组血清PSA、fPSA和EPCA水平较BPH组和健康对照组明显升高(P<0.01),PCa组和BPH组fPSA/PSA水平低于健康对照组(P<0.01);单项指标中,EPCA的灵敏度和特异性最高(78.5%、96.0%),约登指数最大(0.777);建立回归模型Y=logit(P)=-6.906+0.843XPSA-1.402XfPSA+0.271XEPCA,新变量P的AUC为0.935,灵敏度和特异性分别为87.9%和96.0%.结论 PSA、EPCA检测在PCa诊断中具有一定的临床意义,与fPSA联合检测与单项检测相比可以显著提高诊断能力,综合应用Logistic回归和ROC曲线分析有助于提高PCa的诊断效能.     

乳腺癌患者血清IL-6、IL-1β、TNF-α的变化及其临床意义

目的探讨乳腺癌患者血清IL-6、IL-1β、TNF-ɑ的变化及其临床意义。方法应用放射免疫法和免疫比浊法检测66例乳腺癌患者、22例乳腺良性增生患者、62名健康对照者空腹血清IL-6、IL-1β、TNF-ɑ水平。用ROC曲线方法确定各指标的最佳诊断临界值,用Logistic回归分析方法进行三种指标联合分析。结果乳腺癌组血清三种指标水平均高于乳腺良性增生组和健康对照组（P〈0.05）。单独用其中一种指标鉴别诊断的敏感性和特异性均不超过83%,若用Logistic回归分析联合＂IL-6＋IL-1β＋TNF-ɑ＂鉴别诊断乳腺癌和乳腺良性增生的敏感度可以达到96.7%,但特异度仅71.4%。结论血清IL-6、IL-1β、TNF-ɑ可作为乳腺癌鉴别诊断的辅助指标。     

estern印迹杂交分析p53蛋白在人鼻咽癌中的表达及其意义

目的 :检测 10 4例鼻咽癌活检组织和 8例鼻咽粘膜慢性炎症组织中p5 3蛋白的表达。方法 :采用Western印迹法结合免疫沉淀技术。结果 :10 4例鼻咽癌活检组织中有 12例出现突变型p5 3蛋白表达 ,突变检出率为 11.5 4% (12 10 4) ;8例鼻咽慢性炎症活检组织中未见p5 3蛋白的异常表达 (0 8) ;鼻咽癌病人中人IgA VCA抗体滴度≥ 1∶16 0者 ,p5 3蛋白突变检出率为11.80 % (11 93) ,IgA VCA抗体滴度≤ 1∶80者 ,p5 3蛋白突变检出率为 9.10 % (1 11) ,IgA EA抗体滴度阳性者 ,p5 3蛋白突变检出率为 14.10 % (11 78) ,IgA EA抗体滴度阴性者 ,p5 3蛋白突变检出率为 3.80 % (1 2 6 ) ;12例p5 3蛋白异常表达的鼻咽癌病人中有 2例复发。结论 :突变型p5 3基因产物的表达与鼻咽癌EB病毒早期抗原有关 ,p5 3基因在鼻咽癌中可能发挥一定的作用     

Single-assay combination of Epstein-Barr Virus (EBV) EBNA1- and viral capsid antigen-p18-derived synthetic peptides for measuring anti-EBV immunoglobulin G (IgG) and IgA antibody levels in sera from nasopharyngeal carcinoma patients: options for field screening

Assessment of immunoglobulin A (IgA) antibody responses to various Epstein-Barr virus (EBV) antigen complexes, usually involving multiple serological assays, is important for the early diagnosis of nasopharyngeal carcinoma (NPC). Through combination of two synthetic peptides representing immunodominant epitopes of EBNA1 and viral capsid antigen (VCA)-p18 we developed a one-step sandwich enzyme-linked immunosorbent assay (ELISA) for the specific detection of EBV reactive IgG and IgA antibodies in NPC patients (EBV IgG/IgA ELISA). Sera were obtained from healthy donors (n = 367), non-NPC head and neck cancer patients (n = 43), and biopsy-proven NPC patients (n = 296) of Indonesian and Chinese origin. Higher values of optical density at 450 nm for EBV IgG were observed in NPC patients compared to the healthy EBV carriers, but the large overlap limits its use for NPC diagnosis. Using either EBNA1 or VCA-p18 peptides alone IgA ELISA correctly identified 88.5% and 79.8% of Indonesian NPC patients, with specificities of 80.1% and 70.9%, whereas combined single-well coating with both peptides yielded sensitivity and specificity values of 90.1 and 85.4%, respectively. The positive and negative predictive values (PPV and NPV, respectively) for the combined EBNA1 plus VCA EBV IgA ELISA were 78.7% and 93.9%, respectively. In the Indonesia panel, the level of EBV IgA reactivity was not associated with NPC tumor size, lymph node involvement, and metastasis stage, sex, and age group. In the China panel the sensitivity/specificity values were 86.2/92.0% (EBNA1 IgA) and 84.1/90.3% (VCA-p18 IgA) for single-peptide assays and 95.1/90.6% for the combined VCA plus EBNA1 IgA ELISA, with a PPV and an NPV for the combined EBV IgA ELISA of 95.6 and 89.3%, respectively. Virtually all NPC patients had abnormal anti-EBV IgG diversity patterns as determined by immunoblot analysis. On the other hand, healthy EBV carriers with positive EBV IgA ELISA result showed normal IgG diversity patterns. By using EBV IgG immunoblot diversity as confirmation assay for EBV IgA ELISA-positive samples, the sensitivity and specificity for NPC diagnosis increased to 98% and 99.2%, respectively, in the Indonesian NPC samples. The use of these combined methods for seroepidemiological screening studies is proposed.     

Native early antigen of Epstein-Barr virus, a promising antigen for diagnosis of nasopharyngeal carcinoma

The Epstein-Barr virus (EBV) early antigen (EA) complex consists of multiple proteins with relevance for diagnosis of acute, chronic and malignant EBV related diseases, including nasopharyngeal carcinoma (NPC). In a recent study, it was found that the molecular diversity of EBV-specific IgG and IgA antibody responses in NPC patients and demonstrated that these reflect independent B-cell triggering leading to distinct EBV antigen-recognition profiles. The fine-specificity of NPC-related IgG and IgA responses was explored further against defined recombinant and synthetic EBV-EA antigens using immunofluorescence, immunoblot and ELISA techniques and determined their diagnostic value in a large panel of sera from NPC (n = 154), non-NPC tumor patients (n = 133), acute mononucleosis patients (n = 70) and healthy EBV carriers (n = 259). Individual recombinant EBV-EA markers yielded sensitivity/specificity values not exceeding 86%, whereas selected EA-specific peptide epitopes were rather poorly recognized by IgG and IgA antibodies in NPC sera. Surprisingly, we found that a "low salt" native EA-protein extract reproducibly prepared from purified nuclei of EA-induced HH514 cells, and containing characteristic EA(D)-polypeptides, such as p47-54 (BMRF1), p138 (BALF2), p55-DNAse (BGLF5), and p65-TK (BXLF1), but without viral capsid (VCA) or nuclear antigen (EBNA) reactivity, gave highest sensitivity (90.4%) and specificity (95.5%) values for NPC diagnosis in both IgG and IgA ELISA. The data support further the notion that EBV-EA reactive IgG and IgA antibodies in NPC patients are directed against distinct conformational and-in part-linear epitopes on EBV-specific proteins, barely recognized in other EBV-related syndromes. The use of a defined native EBV EA-specific antigen opens the way to further improve serological diagnosis of NPC.     

Epstein-Barr virus serology in the diagnosis of nasopharyngeal carcinoma

The antibody levels to viral capsid antigen (VCA) and early antigen (EA) of Epstein-Barr virus (EBV) in 164 nasopharyngeal carcinoma (NPC) patients from Sarawak, East Malaysia were significantly higher than those in 147 sex, age and ethnically matched healthy controls. As diagnostic markers of NPC, IgG/VCA at reciprocal titers > or =160 was the most sensitive (89%, with 98% specificity), while IgA/EA at > or =5 was the most specific (100%) but the least sensitive (75%). The sensitivity and specificity of IgA/VCA at reciprocal titers > or =10 were 84% and 97%. IgA/VCA has an advantage over IgG/VCA despite the slightly lower sensitivity due to its consistently more distinct fluorescence reaction. The sensitivity and specificity can be marginally improved by a combination of two tests.     

Comparison of three different serological techniques for primary diagnosis and monitoring of nasopharyngeal carcinoma in two age groups from Tunisia

Nasopharyngeal carcinoma (NPC) in Tunisia is characterized by its bimodal age distribution involving juvenile patients of 10-24 years and adult patients of 40-60 years. Three serological techniques were compared for primary diagnosis (N = 117) and post-treatment monitoring (N = 21) of NPC patients separated in two age groups. Immunofluorescence assay (IFA) was used as the "gold standard" for detection of IgG and IgA antibodies reactive with Epstein-Barr virus (EBV) early (EA) and viral capsid (VCA) antigens. Results were compared with ELISA measuring IgG and IgA antibody reactivity to defined EBNA1, EA, and VCA antigens. Immunoblot was used to reveal the molecular diversity underlying the anti-EBV IgG and IgA antibody responses. The results indicate that young NPC patients have significantly more restricted anti-EBV IgG and IgA antibody responses with aberrant IgG VCA/EA levels in 78% compared to 91.7% in elder patients. IgA VCA/EA was detected in 50% of young patients versus 89.4% for the elder group (P < 0.001). Immunoblot revealed a reduced overall diversity of EBV antigen recognition for both IgG and IgA in young patients. A good concordance was observed between ELISA and IFA for primary NPC diagnosis with 81-91% overall agreement. Even better agreement (95-100%) was found for antibody changes during follow-up monitoring, showing declining reactivity in patients in remission and increasing reactivity in patients with persistent disease or relapse. ELISA for IgA anti-VCA-p18 and immunoblot proved most sensitive for predicting tumor relapse. VCA-p18 IgA ELISA seems suitable for routine diagnosis and early detection of NPC complication.     

Evaluation of commercial EBV RecombLine assay for diagnosis of nasopharyngeal carcinoma

BACKGROUND: In recent years a number of Epstein-Barr virus (EBV) proteins were defined as being immunodominant for either IgM, IgG or IgA immune responses, yielding promising markers for diagnostic serology. Specific reactivity patterns to these proteins have been described for infectious mononucleosis (IM), nasopharyngeal carcinoma (NPC), various types of lymphoma, and healthy EBV carriers. OBJECTIVES: To compare the NPC-related diagnostic value of EBV RecombLine test (Mikrogen, Germany) with a standardized immunoblot assay [Fachiroh J, Schouten T, Hariwiyanto B, Paramita DK, Harijadi A, Haryana SM, et al. Molecular diversity of Epstein-Barr virus IgG and IgA antibody responses in nasopharyngeal carcinoma: a comparison of Indonesian, Chinese, and European subjects. J Infect Dis 2004;190:53-62] and to define the diagnostic value of individual EBV marker proteins in a population with high incidence of NPC. RESULT: Sera from Indonesian NPC patients taken at primary diagnosis (n=108) were analyzed for IgG and IgA reactivity and compared with regional healthy blood donors (n=62), non-NPC patient controls (n=10) and IM patients (n=10). Most NPC patients and controls showed strong IgG reactivity to VCA-p18, -p23, and EBNA1, limiting their diagnostic use. Few (<20%) healthy donors and patient controls showed IgG reactivity to EA proteins p47/54 and p138, yielding combined sensitivity/specificity and PPV/NPV values of 92.6%/98.3% and 99.0%/88.1%, for diagnosing NPC. NPC sera showed significantly more EBV reactive IgA antibody (>80% positive) than controls (<10% positive), although being less broadly reactive and significantly less strong compared to IgG. For IgA best results were observed for RecombLine EBNA1 with sensitivity/specificity and PPV/NPV values of 92%/89% and 93.4%/85.9%, respectively. CONCLUSION: In high incidence NPC regions with low incidence IM yet high prevalence of EBV infection, both RecombLine IgG and IgA tests provide a useful alternative to the more complex cell-extract based immunoblot assay as confirmation test for NPC diagnosis in particular when using EA and EBNA1 as discriminators in IgG and IgA testing, respectively.     

Performance of Two Commercially Available Automated Immunoassays for the Determination of Epstein-Barr Virus Serological Status

This study evaluated the performance of two automated Vidas (V) and Liaison (L) immunoassays for Epstein-Barr virus (EBV) serology. The detection of the viral capsid antigen (VCA) IgM, the VCA/early antigen (VCA/EA) IgG, and the Epstein-Barr nuclear antigen (EBNA) IgG was assessed on 526 sera collected for routine EBV testing in immunocompetent subjects. The determination of expected EBV status (186 EBV primary infections, 183 past EBV infections, and 157 EBV-seronegative individuals) was based on results of routine laboratory enzyme immunoassays (EIAs) together with clinical data. The sensitivity and specificity of each individual marker were determined in comparison to the expected EBV status. The agreement between the V and L profiles and the expected EBV status was established through the interpretation of combinations of the different EBV markers. Statistically significant differences between the two tests were found for the specificity of the VCA IgM marker (96.2% for V versus 93.2% for L), the sensitivity of the VCA/EA IgG marker (89% for V versus 94% for L), and the specificity of the EBNA IgG marker (96.5% for V versus 74.2% for L). The results determined for the two assays with respect to overall agreement with the established expected EBV status were not significantly different (89.7% for V versus 88.2% for L), with discrepancies mainly observed in sera referenced as primary infections. These findings demonstrated the similar performances of the Vidas and the Liaison assays for the establishment of an EBV serological status using the VCA, EA, and EBNA markers.