ELISA法检测中国中山市正常人及鼻咽癌患者血清EB病毒抗体水平

目的观察EB病毒抗体 EBNA1/IgA、ZEBRA/IgG、EBNA1/IgG在不同年龄、不同性别的正常人群中的分布,并与鼻咽癌患者进行比较。方法用ELISA法在全自动酶联免疫分析系统上检测468例正常人和16例鼻咽癌患者的血清标本。结果正常人血清各抗体的水平[rD（λ）值]分别为:EBNA1/IgA男性0.726±0.541,女性0.563±0.340,男性高于女性（P<0.001）;ZEBRA/IgG男性0.709±0.480,女性0.829±0.480,女性高于男性（P<0.01）;EBNA1/IgG男性0.781±0.285,女性0.784±0.302,男女之间无显著性差异（P>0.05）。ZEBR/IgG的阳性率随年龄增高而升高,相关系数为0.766（P<0.05）。鼻咽癌患者上述3种抗体的血清水平分别为3.265±1.76、1.408±0.672、1.249±0.317,均明显高于正常人（P<0.01）。结论正常人群中EBNA1/IgA、ZEBRA/IgG、EBNA1/IgG3种抗体的血清水平存在年龄和性别上的差异,但波动在一定范围内;在鼻咽癌的临床诊断和高危人群筛查中有必要根据具体的人群界定不同的阳性临界值;鼻咽癌患者此3种抗体的血清水平均明显升高,对鼻咽癌的诊断有一定意义。     

EB病毒Rta/IgG、EBNA1/IgA、VCA/IgA 及EA/IgA 抗体与鼻咽癌分期的关系

目的探讨EB病毒Rta/IgG、EBNA1/IgA、VCA/IgA及EA/IgA抗体与鼻咽癌临床分期的关系。方法收集211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Rta/IgG、EBNA1/IgA抗体,用免疫酶法检测VCA/IgA及EA/IgA抗体,按92分期法进行分期,分别计算各T、N、M分期及临床分期的各抗体阳性率及抗体水平并进行统计学分析。结果鼻咽癌各T、N、M分期及临床分期组Rta/IgG抗体rA值均无统计学差异(P>0.05)。T1期的EBNA1/IgA抗体rA值明显低于其它T分期,N0期明显低于其它N分期,临床I期明显低于其它临床分期(P<0.05)。VCA/IgA、EA/IgA抗体滴度各N分期及临床分期间比较,差异有统计学意义(P<0.05)。结论EB病毒Rta/IgG抗体表达与鼻咽癌分期无关。EBNA1/IgA抗体在早期鼻咽癌表达水平相对较低。而VCA/IgA、EA/IgA抗体水平与颈部淋巴结转移程度相关,对于评估鼻咽癌临床分期有一定的参考价值。     

EB病毒ZEBRA/IgG抗体检测在鼻咽癌血清学诊断中的应用

目的研究ZEBRA/IgG抗体检测在鼻咽癌血清学检测中的应用。方法应用ELISA法检测鼻咽癌新发病例、鼻咽癌治疗后复诊病例、EB病毒VCA/IgA抗体阳性及阴性正常人群中的ZEBRA/IgG抗体。结果鼻咽癌新发病例ZE-BRA/IgG的阳性率为92.7%,显著高于其他各组,均有极显著性差异(P均<0.01)。治疗后复诊病例,ZEBRA/IgG的阳性率为56.9%,抗体水平较治疗前明显下降。结论ZEBRA/IgG可作为诊断鼻咽癌及预后判断的较好指标。     

EB病毒Rta/IgG、EBNAl/IgA、VCA/IgA及EA/IgA抗体与鼻咽癌分期的关系

目的 探讨EB病毒Rta/lgG、EBNA1/IgA、VCA/IgA及EA/IgA抗体与鼻咽癌临床分期的关系.方法 收集211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Rta/IgG、EBNA1/IgA抗体,用免疫酶法检测VCA/IgA及EA/IgA抗体,按"92分期法"进行分期,分别计算各T、N、M分期及临床分期的各抗体阳性率及抗体水平并进行统计学分析.结果 鼻咽癌各T、N、M分期及临床分期组Rta/IgG抗体rA值均无统计学差异(P>0.05).Tl期的EBNA1/IgA抗体rA值明显低于其它T分期,NO期明显低于其它N分期,临床I期明显低于其它临床分期(P<0.05).VCA/IgA、EA/IgA抗体滴度各N分期及临床分期间比较,差异有统计学意义(P<0.05).结论 EB病毒Rta/IgG抗体表达与鼻咽癌分期无关.EBNA1/IgA抗体在早期鼻嘲癌表达水平相对较低.而VCA/IgA、EA/IgA抗体水平与颈部淋巴结转移程度相关,对于评估鼻咽癌临床分期有一定的参考价值.     

EB病毒EBNA1/IgA与鼻咽癌分期的关系

目的：探讨EB病毒EBNA1/IgA抗体与鼻咽癌临床分期的关系。方法：用酶联免疫吸附法（ELISA）检测211例未经治疗的鼻咽癌患者血清中EBNA1/IgA抗体,按＂92分期法＂进行分期,分别计算各T、N、M分期及临床分期的EBNA1/IgA抗体阳性率及抗体水平并进行统计学分析。结果：鼻咽癌患者的EBNA1/IgA抗体表达与性别和各年龄组没有相关性。EBNA1/IgA抗体rA值与原发灶T分期及颈淋巴结转移N分期差异均有统计学意义（P〈0.05） 与远处转移M分期差异无统计学意义（P〉0.05）。鼻咽癌早期（Ⅰ、Ⅱ期）患者的EBNA1/IgA抗体rA值显著低于晚期（Ⅲ、Ⅳ期）（P〈0.05）。结论：EBNA1/IgA抗体在早期鼻咽癌表达水平相对较低。对于评估鼻咽癌临床分期有一定的参考价值。     

广州地区成年女性EB病毒抗体水平相关性及其年龄分布

目的调查广州地区成年女性血清免疫球蛋白水平和抗EB病毒VCA/EBNA1 IgA和IgG抗体的年龄分布及其相互关系。方法收集广州市309例常规体检的健康女性血清样本,分别采用比浊法和ELISA检测血清免疫球蛋白水平、抗EB病毒VCA/EBNA1 IgA和IgG抗体水平。结果广州地区成年女性抗EB病毒VCA-IgA、抗VCA-IgG及抗EBNA1-IgA、抗EBNA1-IgG的阳性率分别为14.6%、92.2%、3.2%、99.7%。抗VCA-IgA阳性率与年龄呈正相关(r=0.163,P=0.004);抗EBNA1-IgG OD值与年龄呈负相关(r=-0.178,P=0.020)。各抗体水平间的相关性分析显示,抗EBNA1-IgA与血清总IgG水平之间呈负相关(r=-0.154,P=0.008);抗VCA-IgA与抗EBNA1-IgA、抗EBNA1-IgA与抗EBNA1-IgG、血清总IgA与抗VCA-IgA、总IgG与抗VCA-IgG、总IgA与总IgG、总IgM与总IgG水平之间均存在正相关(r分别为0.113、0.241、0.228、0.228、0.433和0.362,P0.05)。结论广州地区成年女性99.7%存在EB病毒既往感染。随着年龄增长,抗VCA-IgA阳性率逐渐增高,而抗EBNA1-IgG OD值则逐渐下降。抗EB病毒抗体水平间存在一定的相关性,有助于鼻咽癌等疾病筛检时联合指标的选择。     

中山市健康成人外周血EB病毒抗体谱的调查研究

目的探讨来自不同地域的人群感染EB病毒的血清学抗体谱是否存在差异。方法从鼻咽癌高发区中山市的健康成人中收集348例中山籍贯原住居民的血清和81例外省籍贯居民的血清,用ELISA法检测血清中的EBNA1-IgA、EBNA1-IgG、VCA-p18-IgA、VCA-p18-IgG、Zta-IgA和Zta-IgG等6种抗体水平,比较不同抗体组合时两类人群样本的抗EB病毒抗体谱的差别。结果当三项IgA类抗体皆为阴性时,外省籍贯阴性率远远大于中山籍贯样本（P=0）;当EBNA1-IgA单阳时,外省籍贯样本阳性率远远高于中山籍贯样本（P=0.001）;当Zta-IgA单项阳性时（P=0.008）,或者Zta-IgA和VCA-p18-IgA同时阳性时（P=0.001）,中山籍贯样本阳性率明显高于外省籍贯样本;而三项IgG类抗体的任意组合,中山籍贯和外省籍贯都无统计学意义。结论中山籍贯原住居民感染的EB病毒常常处于激活状态;相反,外省籍贯的中山居民感染的EB病毒处于潜伏期时表达的EBNA1水平更高。提示中山籍贯原住居民比外省籍贯居民具有更高的风险罹患鼻咽癌。     

EARLY NASOPHARYNGEAL CARCINOMA AMONG lgA/VCA ANTIBODY POSITIVE INDIVIDUALS DETECTED BY ANTICOMPLEMENT lMMUNOENZYMATIC METHOD

The anticomplement immunoenzymatic method (ACIE) is used to detect nasopharyngeal carcinoma in EBV IgA/VCA antibody positive in- dividuals in a high risk area. Carcinoma cells with EBNA were found in 4 0f the 64 antibody positive individuals. Cytological and histological examinations showed that stage I poorly dif- ferentiated carcinomas were in 4 cases. The intervals between the first bleeding for serology and nasopharyngeal carcinoma (NPC) diagnosis were 8-9 months. No further elevation of IgA/VCA antibody appeared during this period. The reisult of this study confirm the value of anticomplement immunoenzyme for detecting early stage NPC.     

鼻咽癌病人多项免疫功能及EB病毒PCR检测的综合研究

为进一步了解ＥＢ病毒与鼻咽癌的关系,全面综合调查鼻咽癌病人的免疫功能,应用ＰＣＲ方法检测鼻咽病人组织块ＥＢ病毒ＤＮＡ,并对处国咽癌病人血清中ＥＢ病毒（ＥＢＶ－ＶＣＡ／ＩｇＡ）抗体、超氧化物歧化酶（ＳＯＤ）活性、脂质过氧化物降解产物（ＭＤＡ）及数种微量元素含量进行测定。结果：从鼻咽癌病人鼻咽部活检组织中成功地检测出ＥＢ病毒ＤＮＡ片断。进一步证实了ＥＢ病毒与鼻咽癌的发生有密切关系。并对鼻咽癌病人血清Ｖ     

Serological diagnosis of nasopharyngeal carcinoma by enzyme linked immunosorbent assay: optimization, standardization and diagnostic criteria

OBJECTIVE To produce an enzyme linked immunosorbent assay (ELISA) to detect antibodies against Epstein Barr virus (EBV) specified nuclear antigen 1 (EBNA 1) and the 18 kD EBV matrix protein, and to determine and optimize its sensitivity and specificity for the diagnosis of nasopharyngeal carcinoma (NPC).

METHODS We used a combination of highly purified glutathione transferase fusion proteins of the 40 kD carboxy domain of EBNA1 and the 18 kD EBV matrix protein for coating ELISA plates. In three separate studies, we tested for IgA antibodies in serum specimens from 28 EBV seronegative donors, 284 EBV seropositive donors and 160 newly diagnosed NPC patients. By comparing the sensitivity and specificity of diagnosis obtained for different cutoff values, we derived several quantitative parameters to evaluate assay performance, establish objective diagnostic criteria which optimize the intrinsic diagnostic capability of the assay and assess the significance of individual test results, respectively. Optimum cutoff optical density (OD) is defined as the cutoff OD where sensitivity of the assay equals its specificity, and resolution of the assay is indicated by the value of sensitivity (or specificity) determined at the optimum cutoff OD. Diagnosis of NPC was achieved by setting a cutoff zone at +/- 20% of this value.

RESULTS All the EBV seronegative donors tested were not reactive, and most of the EBV seropositive donors were weakly reactive, while the majority of NPC patients were moderately or strongly reactive. While the assay was thus shown to be specific for EBV, there was an overlap in the level of these serum antibodies between few individuals of the two latter groups. It was shown that the assay performed equally well in two separate studies conducted under different testing conditions and using different collections of sera in that assay resolution determined on these occasions were 86% and 87% respectively. Diagnosis of NPC can be achieved at the same expected sensitivity of 89% and 83% determined at the lower and upper limits of the cutoff zones, with the corresponding values of specificity being 78% and 91%. It was further shown in the third study that resolution of the assay can be increased to 90% using an assay produced with a higher concentration of the same antigens, and that diagnosis of NPC can be achieved at a higher sensitivity ranging between 86% and 95% at a corresponding specificity of 93% and 86%.

CONCLUSIONS After optimization and standardization, the ELISA can achieve a sensitivity ranging from 86% to 95%, with corresponding specificities of 93% and 86% respectively for the diagnosis of NPC.

     

Serological diagnosis of nasopharyngeal carcinoma by enzyme linked immunosorbant assay:optimization,standardization and diagnostic criteria

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鼻咽癌患者血清中EB病毒四种特异性IgG抗体的测定

用间接免疫荧光法检测了２４例鼻咽癌患者与１１９例正常人血清中ＥＢ病毒４种特异性ＩｇＧ抗体，结果表明，正常人血清中ＥＢ病毒衣壳抗原（ＷＣＡ）的抗体效价为１：１６０以下，而鼻咽癌患者血清抗体效价为ｌ：１６０～１：５１２０，抗体阳性率均为１００％；正常人血清中ＥＢ病毒核抗原（ＥＢＮＡ）的抗体效价为１：８０以下，而鼻咽患者血清抗体价为１：８０～１：２５６０，阳性率均为１００％；正常人血清中ＥＢ病毒早期抗原（ＥＡ）的抗体效价为ｌ：１０，阳性率只有１３．３％，而鼻咽癌患者血清抗体效价为ｌ：２０～１：１２８０，阳性率为１００％；正常人血清中ＥＢ病毒抗原（ＭＡ）的抗体效价为１：１０～１：４０，阳性率为７１％，而鼻咽癌患者血清抗体效价为１：１０～１：３２０，阳性率为１００％．以上４种抗体中．ＶＣＡ、ＥＢＮＡ和ＥＡ抗体的效价和分布在正常人与鼻咽癌患者之间有明显差异（Ｐ＜姜勇男，男，３１岁，讲师．延吉市局子街２１号（１３３０００）０．０ｌ），ＥＡ抗体的阳性率也比正常人明显升高（Ｐ＜０．０ｌ），ＭＡ抗体的效价和阳性率与正常人相比无显著差异（Ｐ＞０．０５）。     

类风湿关节炎患者血清抗EB病毒抗体的临床研究

目的研究ＥＢ病毒感染与类风湿关节炎的临床关系。方法以Ｂ９５－８细胞为抗原底物,用间接免疫荧光法检测血清中抗ＩｇＡＥＢ病毒壳抗原抗体（抗ＩｇＡ／ＶＣＡ抗体）。以免疫印迹法检测血清中抗ＩｇＧＥＢ病毒Ｚ蛋白抗体（抗ＩｇＧ／Ｚ抗体）。结果类风湿关节炎患者血清中抗ＩｇＧ／ＶＣＡ抗体阳性率２１．９％,明显高于正常人及其他风湿病患者。抗ＩｇＧ／Ｚ抗体仅在抗ＩｇＡ／ＶＣＡ抗体阳性时出现。结论抗ＩｇＡ／ＶＣＡ抗体的检测对类风湿关节炎的诊断有一定临床意义。类风湿关节炎患者体内的ＥＢ病毒可能处于激活状态。     

血清急性期蛋白EB病毒IgA／VCA，IgA／EA抗体测定在鼻咽癌诊断中的应用

测定１９０例鼻咽癌（ＮＰＣ）患者血清中急性期蛋白（ＡＰＰＳ）包括：唾液酸（ＳＡ）．α＿１－抗胰蛋白酶（α＿１－ＡＴ），α＿１－酸性糖蛋白（ａα＿１－ＡＧＰ），触珠蛋白（ＨＰ），铁传递蛋白（ＴＦ）和前清蛋白（ＰＡ）及ＥＢ病毒（ＥＢＶ）ＩｇＡ／ＶＣＡ，ＩｇＡ／ＥＡ抗体，并评价ＡＰＰＳ和ＥＢＶＩｇＡ／ＶＣＡ，ＩｇＡ／ＥＡ抗体对鼻咽癌的诊断价值。。将ＡＰＰＳ测定值经逐步判别分析统计学处理，求得判别函数，用以对ＮＰＣ进行判别诊断。结果发现：ＮＰＣ治疗前组ＡＮＰＳＥＢＶＩｇＡ／ＶＣＡ，ＩｇＡ／ＥＡ与临床病理诊断的符合率分别为８１．８８％和５６．５７％，而治疗后未复发组两者的符合率分别为８０．９５％和３３．３３％。结果表明：对ＮＰＣ的诊断和随访ＡＰＰＳ优于ＥＢＶＩｇＡ／ＶＣＡ，ＩｇＡ／ＥＡ抗体。     

Detection and analysis of anti-latent membrane protein 2A antibodies in the sera of patients with Epstein-Barr virus associated malignancies

Background Epstein-Barr virus (EBV) associated malignancies with a Type Ⅱ latency gene expression pattern, such as Hodgkin‘s disease, and nasopharyngeal carcinoma (NPC) , frequently express the EBV antigen latent membrane protein 2A (LMP2A). We expected to establish a highly expressing LMP2A yeast cell strain and get the high quality LMP2A protein, which was used for detection, analysis and characterization of its antibodies in various patients‘ sera of EBV associated malignancies. Methods The plasmid pPICZαA-LMP2A containing the full length of LMP2A cDNA was constructed and transformed to Pichia pastoris GS115 to express LMP2A protein. After fermentation and purification, the LMP2A protein was used as an antigen to detect anti-LMP2A antibodies (Abs) in the sera of patients with EBVassociated malignancies in enzyme linked immunosorbent assay (ELISA) or Western-blot. Results LMP2A was expressed successfully with an expected molecular weight of approximately 54 kD and Abs to LMP2A were strikingly specific to NPC. Two-thirds or more sera from NPC patients were positive for antiLMP2A immunoglobulin G (IgG) Abs. The antibodies were absent from the sera of other EBV-associated diseases except a small fraction of the gastric carcinoma. Comparing anti-viral capsid Ags (VCA) IgA and LMP2A IgA titers in the sera from 76 NPC patients, only 55% were positive for anti-LMP2A IgA Abs while 70% were positive for anti-VCA IgA. However, we found that 3 sera negative for VCA IgA were positive for LMP2A IgA. Conclusion The results suggested the potential significance of LMP2A specific Abs for the diagnosis of EBVassociated malignancies, especially NPC.     

Epstein-Barr virus infection in nasopharyngeal lymphoid hyperplasia

Ｏｂｊｅｃｔｉｖｅ　ＴｏｄｅｔｅｃｔｗｈｅｔｈｅｒＥｐｓｔｅｉｎＢａｒｒｖｉｒｕｓ（ＥＢＶ）ｈａｒｂｏｒｓｉｎｎａｓｏｐｈａｒｙｎｇｅａｌｌｙｍｐｈｏｉｄｈｙｐｅｒｐｌａｓｉａ（ＮＰＬＨ）ｗｈｉｃｈｉｓｆｒｅｑｕｅｎｔｌｙｔｏｂｅｓｅｅｎｉｎＧｕａｎｇｚｈｏｕ,ａｈｉｇｈｉｎｃｉｄｅｎｃｅａｒｅａｏｆｎａｓｏｐｈａｒｙｎｇｅａｌｃａｒｃｉｎｏｍａ（ＮＰＣ）,ａｎｄｔｏｅｘｐｌｏｒｅｔｈｅｒｅｌａｔｉｏｎｂｅｔｗｅｅｎＮＰＬＨａｎｄｄｅｖｅｌｏｐｍｅｎｔｏｆＮＰＣＭｅｔｈｏｄｓ　Ｔｗｅｎｔｙ…     

鼻咽癌患者血清EB病毒DNA片段的检出及临床意义

目的：探讨鼻咽癌患者血清Epstein-Barr病毒(EBV)DNA片段检出率及其和EBV衣壳抗原IgA类抗体(VCA／IgA)的关系。方法：聚合酶链反应(PCB)加限制性内切酶酶切技术检测疱疹病毒的DNA，酶免疫组化技术检测EBV VCA／IgA抗体。结果：33例鼻咽癌患者中，抗体阳性率88％，DNA阳性率55％；对照组120例，仅1例检出EBV DNA片段。鼻咽癌患者中EBV DNA片段的检出率显著性高于对照组，但EBVDNA片段的检出率低于EBV VCA／IgA抗体的检出率。结论：进一步证实EBV与鼻咽癌有密切关系；EBV DNA片段及EBV VCA／IgA抗体检测，有利于鼻咽癌的辅助诊断。     

益气解毒颗粒对鼻咽癌高危人群EB病毒感染活性抑制作用的临床观察

为观察益气解毒颗粒对鼻咽癌高危人群EB病毒感染活性的抑制作用.将111例鼻咽癌高危患者随机分为2组,分别予益气解毒颗粒及鼻咽清毒剂,连用3月,观察其对壳抗原抗体(IgA/VCA)和早期抗原抗体(IgA/EA)滴度的影响.结果:治疗组IgA/VCA由21.485±2.956降至3.678±3.821,IgA/EA由6.987±1.575降至1.771±2.407,显效率51.72%,总有效率87.93%;对照组则分别由21.793±3.094降至15.139±3.375,7.598±1.732降至5.438±2.329,显效率7.55%;总有效率24.53%,组间差异均为P＜0.01.结论:益气解毒颗粒对鼻咽癌高危者EB病毒感染活性有很强的抑制作用.     

鼻咽癌患者外周血白细胞及血清中EB病毒DNA的检测

目的探讨鼻咽癌患者外周血白细胞及血清中EB病毒DNA的分布及其与鼻咽癌的关系.方法采用多聚酶链反应检测24例鼻咽癌患者(实验组)及10例头颈部其它部位肿瘤(对照组-1)和10例慢性鼻咽炎患者(对照组-2)外周血白细胞及血清中EB病毒的DNA.同时采用免疫酶法检测以上各组患者血清中EB病毒VCA-IgA抗体的效价.结果发现实验组中10例患者外周血白细胞中EB病毒DNA为阳性,阳性率为41.67%(10/24).对照组-1阳性率为10%(1/10),对照组-2患者外周血白细胞中EB病毒DNA均为阴性.实验组与对照组-2比较阳性率差异有显著性(P＜0.05).在以上三组患者血清中EB病毒DNA均为阴性,三组间无差异.实验组VCA-IgA与对照组-1,对照组-2分别比较阳性率其差异均有显著性(P＜0.01).结论有些鼻咽癌患者外周血白细胞中可检测出EB病毒DNA.血清中未能检测出EB病毒DNA.鼻咽癌患者外周血白细胞EB病毒DNA的存在与血清VCA-IgA的效价之间有一定的关系.     

广西五地区338868人鼻咽癌普查比较分析

目的　提高全人口鼻咽癌早诊率。方法　对广西五地区３３８８６８ 人进行鼻咽癌血清学检查,对检出患者进行组织学病理确诊。结果　检出 Ｅ Ｂ病毒壳抗原的免疫球蛋白 Ａ（ Ｖ Ｃ Ａ Ｉｇ Ａ）抗体阳性９３６７ 人,早期抗原的免疫球蛋白 Ａ（ Ｅ Ａ Ｉｇ Ａ）阳性３０６ 人。检出鼻咽癌１１３ 例,各地区早诊率为６６７％ ～１００％ 。结论　证明 Ｅ Ｂ病毒与鼻咽癌关系密切,血清学普查可以检出鼻咽癌和早期鼻咽癌。