[1]张玲,刘杰,王臣玉,等.EB 病毒Rta 优势表位抗原的克隆表达及在应用ELISA 诊断鼻咽癌中的价值[J].现代检验医学杂志,2020,35(05):1-4,12.[doi:10.3969/j.issn.1671-7414.2020.05.001]
　ZHANG Ling,LIU Jie,WANG Chen-yu,et al.Cloning and Expression of Dominant Epitope Antigen of Epstein-Barr Virus Rta Protein and Its Application in the Diagnosis of Nasopharyngeal Carcinoma by ELISA[J].Journal of Modern Laboratory Medicine,2020,35(05):1-4,12.[doi:10.3969/j.issn.1671-7414.2020.05.001]
点击复制

EB 病毒Rta 优势表位抗原的克隆表达及在应用ELISA 诊断鼻咽癌中的价值()
分享到：

《现代检验医学杂志》[ISSN:/CN:]

卷:
第35卷
期数:
2020年05期
页码:
1-4,12
栏目:
论著
出版日期:
2020-11-01

文章信息/Info

Title:
Cloning and Expression of Dominant Epitope Antigen of Epstein-Barr Virus Rta Protein and Its Application in the Diagnosis of Nasopharyngeal Carcinoma by ELISA
文章编号:
1671-7414（2020）05-001-05
作者:
张玲1; 刘杰2; 王臣玉2; 张守信2; 杨丽萍2; 危利1; 王超男1; 焉丽波3;  宋少峰3; 陈蒙蒙3; 冯晓燕1; 张贺秋1
1. 东方海洋（北京）医学研究院，北京 100071；2. 烟台毓璜顶医院，山东烟台264000； 3. 艾维可生物科技有限公司，山东烟台264000
Author(s):
ZHANG Ling1; LIU Jie2; WANG Chen-yu2; ZHANG Shou-xin2; YANG Li-ping2; WEI Li1; WANG Chaonan1 ; YAN Li-bo3; SONG Shao-feng3; CHEN Meng-meng3; FENG Xiao-yan1; ZHANG He-qiu1
1. Medical Institute of Oriental Ocean (Beijing), Bijing 100071, China; 2. Yantai Yuhuangding Hospital, Shandong Yantai 264000, China; 3. Avioq Biotechnology Co. LTD, Shandong Yantai 264000, China
关键词:
EB 病毒; 立即早期蛋白Rta; 优势表位抗原; 鼻咽癌
分类号:
R739.62；R730.43
DOI:
10.3969/j.issn.1671-7414.2020.05.001
文献标志码:
A
摘要:
目的　制备高活性EB 病毒立即早期蛋白Rta 优势表位抗原，并初步评价该抗原在鼻咽癌（nasopharyngeal carcinoma, NPC）诊断中的价值。方法　分析Rta 氨基酸序列，选取抗原优势表位Rta（301~440aa），合成目的基因， 连接pBVGST-6His 载体，构建重组pBV-Rta 表达质粒，进行诱导表达获得纯化抗原。采用间接ELISA 初步评价优势表 位抗原在鼻咽癌诊断中的价值，以纯化的pBV-Rta 优势表位抗原为包被抗原，抗人IgA，IgG 和IgM 为二抗，检测由烟 台毓璜顶医院提供的50 例经临床病理确诊的NPC 患者血清样本和90 例健康体检者血清样本。结果　获得了高效原核 表达的pBV-Rta 优势表位抗原(301~440aa)，经过小样本验证，确定pBV-Rta 具有良好的抗原活性和特异度。放大样本 进行检测，基于Rta 优势表位抗原的Rta-IgG 间接ELISA 方法检测灵敏度和特异度分别为73.08% 和95.79%。Rta-IgG 检测的AUC 值为0.860。结论　Rta 优势表位抗原pBV-Rta 是优选的可以用于Rta-IgG 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。
Abstract:
Objective　To prepare the dominant epitope antigen of Rta of Epstein Barr virus (EBV) immediate early protein, and evaluate its value in the diagnosis of nasopharyngeal carcinoma(NPC). Methods　The amino acid sequence of Rta was analyzed, and the dominant epitope antigen of Rta (301~440aa) was selected to synthesize the target gene. An expression-construct for Rta (301~440aa) was engineered by inserting the corresponding DNA into a pBVGST-6His plasmid. The recombinant pBV-Rta expression plasmid was induced expression to obtain the purified antigen.The value of the dominant epitope antigen in the diagnosis of NPC was preliminarily evaluated by indirect ELISA. The purified pBV-Rta dominant epitope antigen was used as the encapsulated antigen, and the anti-human IgA, IgG and IgM were used as the secondary antibody. The sensitivitiy and specificitiy of pBV-Rta were detected in serum of 50 NPC patients confirmed by clinicopathology and 90 healthy patients tested by Yantai Yuhuangting Hospital. Results　The domonant epitope antigen (301~440aa) of pBV-Rta was highly prokaryotic expressedin E. coli. After small sample verification, it was determined that pBV-Rta had good antigenic activity and specificity. The sensitivitiy and specificitiyof Rta-IgG were 73.08% and 95.79%, respectively, by indirect ELISA based on Rta domonant epitope antigen,and the AUC value of Rta-IgG was 0.860. Conclusion　The Rta dominant epitope antigen pBV-Rta is the preferred antigen that can be used for Rta-IgG detection. The indirect ELISA detection methods of Rta-IgG established using Rta dominant epitope antigen can be used to distinguish NPC patients from healthy controls.

参考文献/References:

[1] EPSTEIN M A, ACHONG B G, BARR Y M. Virus particles in cultured lymphoblasts from Burkitt’s lymphoma[ J]. The Lancet, 1964, 283(7335): 702-703.
[2] GUIGAY J. Advances in nasopharyngeal carcinoma[J]. Current Opinion in Oncology, 2008, 20(3): 264-269.
[3] CHUA M L K, WEE J T S, HUI E P, et al. Nasopharyngeal carcinoma[J]. Lancet, 2016, 387(10022): 1012-1024.
[4] CAO Sumei, SIMONS M J, QIAN Chaonan. The prevalence and prevention of nasopharyngeal carcinoma in China[J]. Chinese Journal of Cancer, 2011, 30(2): 114-119.
[5] AI Ping, WANG Tao, ZHANG Hong, et al. Determination of antibodies directed at EBV proteins expressed in both latent and lytic cycles in nasopharyngeal carcinoma[ J]. Oral Oncology, 2013, 49(4): 326-331.
[6] FENG Ping, REN E C, LIU Dingxiang, et al. Expression of epstein–barr virus lytic gene BRLF1 in nasopharyngeal carcinoma: potential use in diagnosis[J]. Journal of General Virology, 2000, 81(10): 2417-2423.
[7] 孙畅, 路顺，郞锦义．EB 病毒感染与鼻咽癌诊治的研 究进展[J]. 肿瘤预防与治疗, 2018, 31（5）: 352-357.
SUN Chang, LU Shun, LANG Jinyi. Research progress of Epstein-Barr virus infection and the diagnosis and treatment of nasopharyngeal carcinoma[J].J Cancer Control Treat, 2018, 31(5): 352-357.
[8] 陈刚．EB 病毒感染的实验室诊断研究进展[J]．中 国热带医学，2018，18（3）：289-293．
CHEN Gang. Research progress of laboratory diagnosis of Epstein-Barr virus infection[J]. China Tropical Medicine, 2018, 18（3）: 289-293．
[9] 何流，张雷．鼻咽癌患者血清EB 病毒Rta-IgG、 VCA-IgA、EA-IgA 抗体与其临床表现的关系[J]． 解放军医药杂志, 2020, 32（2）: 44-47．
HE Liu, ZHANG Lei. Correlations between serum EB virus ,Rta-IgG，VCA-IgA and EA-IgA antibodies with clinical manifestations in patients with nasopharyngeal carcinoma [J]. Medical &Pharmaceutical Journal of Chinese People’s Liberation Army, 2020, 32(2): 44-47.
[10] 任军，张晓梅，张晓光，等．以Rtac2/3 为抗原用 于鼻咽癌病人检测的初步研究[J]. 中华微生物学和 免疫学杂志, 2006, 26（11）: 1057-1059．
REN Jun, ZHANG Xiaomei, ZHANG Xiaoguang, et al. Studies on antibody response to recombinant Rta protein in patient with nasopharyngeal carcinoma [J]. Chinese Journal of Microbiology and Immunology, 2006, 26(11): 1057-1059.
[11] 祝元雪，李晓江．EB 病毒Rta 蛋白在鼻咽癌早期 诊断中的研究进展[J]．现代肿瘤医学, 2013, 21（12）: 2844-2846.
ZHU Yuanxue, LI Xiaojiang. Research of EB virus Rta protein in the early diagnosis of nasopharyngeal carcinoma [J]. Journal of Modern Oncology, 2013, 21(12): 2844-2846.
[12] 赵新星, 张龙城. 鼻咽癌与EB 病毒立即早期蛋白的 应用进展[J]. 临床耳鼻咽喉头颈外科杂志, 2016, 30 （13）: 1078-1083．
ZHAO Xinxing, ZHANG Longcheng. Application progress of nasopharyngeal carcinoma and immediate early proteins in EB virus [J]. Journal of Clinical Otorhinolaryngology Head and Neck Surgery, 2016, 30(13): 1078-1083.
[13] 李晓华，蒙以良，漆光紫，等．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[J]. 现代免疫学, 2013, 33（2）: 142- 145.
LI Xiaohua, MENG Yiliang, QI Guangzi, et al. Comparison between 2008 clinical stages of nasopharyngeal carcinoma and Epstein-Barr virus antibodies positive rate of EBNA1/IgA, Zta/IgG and Rta/IgG the zhuang nationality [J]. Current Immunology, 2013, 33(2): 142- 145.
[14] FENG P, CHAN S H, SOO M Y, et al. Antibody response to Epstein-Barr virus Rta protein in patients with nasopharngeal carcinoma[J]. Cancer, 2001, 92(7): 1872-1880.

相似文献/References:

[1]王巧祎,王宁宁,刘晴晴,等.EB 病毒BART miRNAs 在胃癌中的作用机制研究进展[J].现代检验医学杂志,2023,38(05):199.[doi:10.3969/j.issn.1671-7414.2023.05.038]
　WANG Qiaoyi,WANG Ningning,LIU Qingqing,et al.Research Progress on the Mechanism of Epstein-Barr Virus BART miRNAs in Gastric Cancer[J].Journal of Modern Laboratory Medicine,2023,38(05):199.[doi:10.3969/j.issn.1671-7414.2023.05.038]
[2]周林颖,刘 兴,伍 勇,等.HIV/AIDS 患者并发EBV 和HCMV 感染临床免疫学特征及影响因素分析[J].现代检验医学杂志,2024,39(02):91.[doi:10.3969/j.issn.1671-7414.2024.02.017]
　ZHOU Linying,LIU Xing,WU Yong,et al.Clinical Immunological Characteristic and Influencing Factors Analysis of HIV/AIDS Patients Infected with EBV and HCMV[J].Journal of Modern Laboratory Medicine,2024,39(05):91.[doi:10.3969/j.issn.1671-7414.2024.02.017]

备注/Memo

备注/Memo:
基金项目：山东省重大科技创新工程项目（2019JZZY011018）。
作者简介：张玲（1989-），女，硕士研究生，从事分子生物学及蛋白纯化，E-mail：ling808818@126.com。 　　　　　刘杰，硕士研究生，副主任技师，从事分子遗传学检测，E-mail：liulocus@126.com，与第一作者同等贡献。
通讯作者：张贺秋，E-mail：zhangheqiu2004@126.com。
更新日期/Last Update: 2020-10-30