酶联免疫吸附检测鼻咽癌方法的建立

目的：探讨检测鼻咽癌的酶联免疫吸附（ＥＬＩＳＡ）方法。方法：以正丁酸和巴豆油诱导Ｒａｊｉ细胞表达的ＥＢ病毒早期蛋白为抗原，用ＥＬＩＳＡ方法检测血清中抗ＥＢ病毒相关早期蛋白的ＩｇＧ抗体水平（吸光度Ａ４９０）。其中鼻咽癌４５４例，肝癌４０例，肺癌２３例，肠癌２０例，乳腺癌２３例，卵巢癌１６例，头颈肿瘤１５例和健康人５２４例。结果：在所有被检血清中鼻咽癌血清Ａ值最高（０．３３３±０．１００），（Ｐ＜０．００１）。以正常人９５％特异性Ａ４９０＜０．１８为界，ＥＬＩＳＡ方法诊断鼻咽癌的敏感性为８９％（４０５／４５４）特异性为９４％（４９３／５２４）。结论：用来自Ｒａｊｉ细胞内的ＥＢ病毒早期蛋白为抗原建立的ＥＬＩＳＡ方法可以应用于鼻咽癌的血清学诊断。     

EB病毒VCA－IgA抗体微量检测法

ＥＢ病毒ＶＣＡ－ＩｇＡ抗体微量检测法陈于东关键词：ＥＢ病毒抗体，微量法，鼻咽癌鼻咽癌患者血清ＥＢ病毒壳抗原的免疫球蛋白Ａ抗体（ＶＣＡ－ＩｇＡ）阳性率高达９０％以上［１，２］，除了可作为临床诊断、观察疗效的重要指标外，也应用于普查。常规方法需抽取静脉血...     

正常人群痘苗病毒抗体的测定

用ＥＬＩＳＡ方法检测了１５５例不同年龄组正常人血清中的痘苗病毒ＩｇＧ抗体，结果如下１２岁以下儿童全部为阴性，１３岁以上各年龄组抗体阳性率为３５．０％－８０．０％，总阳性率为３８．７％；在各阳性年龄组中，抗体效价分布在１：２００－１：１６００之间，几何平均效价为１：２８３，分布在１：２６９－１：２９５之间。     

SLE患者与EB病毒感染的研究

采用免疫酶标法测定２０例ＳＬＥ患者及２０例正常人血清中ＥＢ病毒壳抗原抗体ＩｇＡ（ＥＢＶＣＡ－ＩｇＡ）。结果显示：ＳＬＥ患者ＥＢＶＣＡ－ＩｇＡ阳性率轻正常人为高（Ｐ＜０．０５）。提示ＳＬＥ发病与ＥＢ病毒感染有一定关系。     

321例EB病毒抗体在多种疾病的分布

采用免疫酶法检测３２１例患者血清的抗ＥＢ病毒壳抗原抗体（ＥＢ－ＶＣＡ－ＩｇＡ）结果，鼻咽癌患者ＥＢ－ＶＣＡ－ＩｇＡ阳性率为７５％，其他各系统也都可以不同的阳性率，运用ＥＬＩＳＡ法检测了８８例疾患的抗ＥＢ病毒的早期抗体，抗ＥＢ病毒核抗原抗体（ＩｇＧ／Ｍ）提示血液系统，消化道系统的部分肿瘤及乳癌，甲状腺癌等与ＥＢ病毒有一定的关系。     

鼻咽癌放疗前后EB病毒抗体滴度的变化

鼻咽癌放疗前后ＥＢ病毒抗体滴度的变化李明周祖金（广西区第二人民医院放疗科桂林市５４１００２）关键词鼻咽癌；ＥＢ病毒；抗体鼻咽癌患者血清中常可测出高滴度的抗ＥＢ病毒多种抗原抗体，其中ＩｇＡ抗体尤其是ＩｇＡＶＣＡ高度特异，其水平与病情呈正相关［１］，为...     

14岁以下正常人群B19病毒抗体的测定

用间接免疫荧光法检测了延边地区１４岁以下不同年龄组的正常人群血清中抗Ｂ１９病毒抗体。结果如下：抗体阳性率为１２．９％，各年龄组抗体阳性率分布不均，主要在７．５％～２２．５％之间，几何平均效价为１：１９，各年龄组的几何平均效价在１：１６～１：２５之间；抗体效价分布在１：１０～１：８０之间，主要在１：４０以下，朝鲜族和汉族的病毒抗体阳性率分别为９．７％和１６．０％，几何平均效价分别为１：２１和１：１８．统计结果表明，各年龄组之间及朝鲜族和汉族之间，抗体阳性率和几何平均效价均无显著性差异（Ｐ＞０．０５）．     

柯萨奇β组病毒感染性心肌炎血清学分型及其临床意义

采用ＥＬＩＳＡ法检测２３例病毒性心肌炎和６６例健康人血清ＣＢＶＩ－６型多价和单价抗体。结果心肌炎组柯萨奇β组病毒（ＣＢＶ）多介抗体阳性率５２．２％（１２／２３），健康人组为２５．５％（１７／６６），两者差异显著（Ｐ〈０．０５）；８３．３％（１０／１２）ＣＢＶ感染性心肌患者的单价抗体水平〉１：１２８００，以该效价作为阳性判断值，则ＣＢＶ２，ＣＢＶ３和ＣＢＶ５单价抗体阳性率分别为２０％（２／１０，５０     

11种分枝杆菌抗原用于肺结核和肺癌血清学鉴别诊断的效果比较

目的：对１１种分枝杆菌抗原制备物用于肺结核和肺癌血清学鉴别诊断的效果进行评估。方法：采用ＥＬＩＳＡ技术分别测定１０４例肺结核患者，１０４例肺癌患者和５２例正常人血清中抗１１种分枝杆菌抗原制备物的抗体水平。结果：淋巴分枝杆菌菌体超声抗原Ｂ６ｂ检测肺结核血清抗体阳性率为８２％，显著高于抗ＰＰＤ抗体的阳性率６２％（Ｐ＜０．０５）。Ｂ６ｂ抗原检测肺癌患者血清抗体的阳性率为１．９％，远低于抗ＰＰＤ抗体的阳性检出率１５．４％（Ｐ＜０．０１）。结论：分枝杆菌抗原制备物的菌种来源、细胞培养方法和抗原制备方法对其用于血清学鉴别诊断的灵敏度和特异性，都有重要影响。其中，Ｂ６ｂ抗原用于肺结核和肺癌的鉴别诊断效果最好     

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。     

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。     

ANTICOMPLEMENT IMMUNOENZYMATIC METHOD OF DETECTING EPSTEIN-BARR NUCLEAR ANTIGEN IN NASO- PHARYNGEAL CARCINOMA CELLS AND NORMAL EPI- THELIAL CELLS

Exfoliated cells collected by negative pres- sure suction from the nasopharynx of patients with poorly differentiat.ed and undifferentiated cell carcinoma and of suspect nasopharyngeal {:arcinoma (NPC) patients were examined for Epstein-Barr nuclear antigen (EBNA) by anti- complement immunoenzymatic method (ACIE). All 79 NPC patients had EBNA-positive carcino- ma cells, while the positive rate of cytologic and histologic examination was only 87.3To and 91.1"z. No EBNA was found in tumor cells from pa- tients with head and neck malignant and benign tumors other than NPC, nor was it found in nasopharyngeal epithelial cells of dead fet.uses. 6 persons with EBNA-positive cells detected by this method, with no carcinoma detected by cytology or histology, were reexamined regularly. Carcinoma cells were detected from l week to 3 months later in 4. This shows that this ACIE method is specific and sensitive and can be used for early NPC detection, especially in combina- tion with serologic detection of Epstein-Barr virus (EBV) antibody. Besides finding EBNA in nasopharyngeal carcinoma cells, it can also be found in normal ciliated columnar epithelial cells and hyperplas- tic cells of the nasopharynx.     

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。     

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。     

系统性红斑狼疮患者抗Sm抗体与EB病毒核抗原-2表达的相关分析

目的 探讨EB病毒(EBV)阳性系统性红斑狼疮(SLE)患者EBV核抗原-2(EBNA2)表达与抗Sm抗体的相关性.方法 SLE患者44例,正常人43名作为对照.①采用PCR-Southern杂交技术检测两组外周血单个核细胞中EBV特异性DNA片段, EBV阳性患者进行RT-PCR和 Southern杂交检测EBNA2的表达.②ELISA法检测EBV特异性IgM抗体.③免疫印迹法检测血清可提取核抗原(ENA)抗体.结果 ①SLE组32例EBV阳性,其中EBNA2 mRNA阳性20例(活动期9例,稳定期11例),EBNA2 mRNA阳性率在活动期和稳定期患者差异无显著性(P＞0.05).②SLE组EBV特异性IgM抗体阳性率为18.18%(8/44),对照组均为阴性,两组有显著性差异(P＜0.05).③SLE抗Sm抗体检出率与EBNA2 mRNA检出率呈正相关(r=0.4107,P＜0.05).结论 EBNA2 mRNA表达可能在一定程度上参与SLE的发病和病情进展.     

EB病毒核抗原1优势表位区段抗原的克隆表达及其在鼻咽癌诊断中的应用

目的制备高活性EB病毒核抗原1(Epstein-Barr virus nuclear antigen 1, EBNA1)的优势表位区段抗原,并初步评价该抗原在鼻咽癌诊断中的应用价值。方法利用生物学软件BIOSUN分析EBNA1抗原的B细胞表位分布,并从中选取含有优势表位的抗原区段。然后利用分子生物学技术构建含有优势表位区段序列的原核表达质粒并进行表达,获得纯化优势表位抗原,采用间接酶联免疫吸附试验(ELISA)初步评价优势表位区段抗原的检测性能。结果成功构建含有优势表位抗原区段序列的原核表达质粒,获得了可溶性表达的EBNA1优势表位区段抗原NA-2(401~641 aa)。通过间接ELISA法对小量样本进行验证,确定NA-2抗原针对EBNA1-IgA抗体具有较高的抗原活性和特异性,较EBNA1-IgG和EBNA1-IgM抗体而言,能够较好地区分鼻咽癌(nasopharyngeal carcinoma, NPC)和健康对照样本。放大样本量进行检测,基于NA-2抗原的EBNA1-IgA抗体间接ELISA方法检测灵敏度和特异性分别为90.38%和91.58%,EBNA1-IgA抗体检测的AUC值为0.942。结论获得EBNA1优势表位区段抗原NA-2,该抗原是优选的可以用于EBNA1-IgA检测的抗原,利用该抗原所建立的间接ELISA检测方法可以很好地区分鼻咽癌患者和健康对照。     

Epstein-Barr virus antibodies in patients with ataxia-telangiectasia and other immunodeficiency diseases

An unusual antibody response to the Epstein—Barr virus (EBV) has been noted in patients with ataxia—telangiectasia. Of a group of 16 such patients 8 were found to have antibodies in their serum to the EBV viral capsid antigen (VCA), and 4 of them also had antibodies to the EBV early antigen (EA); antibodies to the nuclear antigen (EBNA), however, were absent in 3 of the 8. The antibody pattern persisted for more than 2 years in the patients available for follow-up study. In comparison, of 24 patients with various other immunodeficiency syndromes 9 were found to have EBV-VCA antibodies in their serum, but none of the 9 had EA antibodies and 3 lacked EBNA antibodies. Two other groups of subjects, all of whom had EBV-VCA and EBNA antibodies in their serum late after an EBV infection, were also studied; 82 had infectious mononucleosis and 55 were healthy and had no such history. EA antibodies were detected in 45 of the first group during the acute phase of the illness but persisted in only 6 of the 68 who were followed up for more than 2 years, and they were detected in only 7 of the second group.

All eight lymphoblastoid cell lines established from the peripheral blood of the four patients with ataxia—telangiectasia who are still available for follow-up study express EBV-VCA, whereas most similar cell lines established from normal individuals express only EBNA. In two of these patients cell-mediated immunity, as assessed from lymphocyte transformation induced by mitogens, was markedly decreased but autologous cell-mediated immune regression of EBV-induced transformation of B (bone-marrow-derived)-lymphocytes was normal. The percentage of T (thymus-derived)-helper cells was greatly decreased in two of the three patients in whom it was measured, and the percentage of T-suppressor cells was greatly increased in one of them, but the percentage of total T-lymphocytes was within normal limits in all three.

The possible significance of these findings — in particular, the persistence of EA antibodies and the diminished restriction of expression of EA — in the late development of tumours after an EBV infection in patients with ataxia-telangiectasia deserves careful attention. Finally, the apparent correlation between immunoglobulin deficiency and poor or absent EBNA antibody response warrants further study.

     

EA-IgA抗体检测对鼻咽癌诊断价值

应用免疫酶标记抗体间接法同时检测鼻咽癌病人98例(放疗前63例,放疗后35例)和对照组51例血清中EA-IgA、VCA-IgA抗体。结果表明,治疗前鼻咽癌病人的VCA-IgA抗体阳性率较高,而EA-IgA抗体更具有特异性,两者联合检测,可以互相补充,提高诊断可靠性,对早期发现鼻咽癌具有重要价值。结果还显示,鼻咽癌放疗后EA-IgA抗体阳性率明显下降,一般放疗后三年以上者此抗体转阴,复发时又呈阳性,提示对鼻咽癌放疗三年后EA-IgA阳性者应给以重视,可能预示肿瘤残存或复发。     

抗鼠科肉瘤病毒癌基因同源物B1抗体的检测及其在类风湿关节炎诊断中的意义

目的:了解抗鼠科肉瘤病毒癌基因同源物B1(v-raf murine sarcoma viral oncogene homologue B1,BRAF)抗体在类风湿关节炎(rheumatoid arthritis,RA)诊断中的临床意义。方法:纳入RA患者112例、健康对照73例、其他风湿病患者112例,以重组人BRAF蛋白为抗原,采用酶联免疫吸附法(enzyme-linked immunosorbent assays,ELISA)检测患者血清中的抗BRAF抗体水平,并收集RA患者的临床资料,采用SPSS 13.0软件行统计学分析。结果:(1)抗BRAF抗体的阳性率在RA患者组为53.6%,明显高于正常对照组(4.1%,P<0.01)和除骨关节炎(34.2%)组以外的其他风湿病组(强直性脊柱炎11.8%,系统性红斑狼疮20.1%,原发性干燥综合征17.9%,P均<0.01);RA患者血清中抗BRAF抗体滴度也明显高于正常对照组和其他风湿病组(P均<0.01)。(2)抗BRAF抗体对RA诊断的敏感度和特异度分别为53.6%和84.3%。(3)类风湿因子、抗环瓜氨酸抗体、抗角蛋白抗体和抗核周因子抗体阴性RA患者的抗BRAF抗体阳性率分别为52.6%、38.2%、30.3%和31.0%。(4)RA患者的抗BRAF抗体水平与患者的年龄、病程、C反应蛋白水平呈负相关。结论:抗BRAF抗体有助于RA的诊断,并可作为其他自身抗体的补充。