吉林大学学报(医学版) ›› 2013, Vol. 39 ›› Issue (2): 400-404.doi: 10.7694/jldxyxb20130248

• 基础研究 • 上一篇    下一篇

双抗体间接夹心ELISA法检测血清MUC1蛋白方法的建立及其在肺癌诊断中的应用

何秋立1,张 旭2,王 娟2,翟瑞萍2,乔彩霞1,孙霞霞2,倪维华2,高素君1,台桂香2   

  1. 1. 吉林大学第一医院肿瘤中心,吉林 长春 130021;2. 吉林大学白求恩医学院
    免疫学教研室,吉林 长春 130021
  • 收稿日期:2012-09-27 出版日期:2013-03-28 发布日期:2013-03-26
  • 通讯作者: 高素君(Tel:0431-88783455,E-mail:sujung1963@sina.com); 台桂香(Tel:0431-85619403,E-mail:taiguixiang@163.com) E-mail:sujung1963@sina.com;taiguixiang@163.com
  • 作者简介:何秋立(1987-),女,河南省新乡市人,在读医学硕士,主要从事肺癌及白血病的基础与临床研究。
  • 基金资助:

    国家自然科学基金资助课题(30972782);吉林省自然科学基金资助课题(201115049);“重大新药创制”科技重大专项资助课题(2011ZX09102-001-36)

Establishment  of indirect sandwich ELISA  methodfor serum MUC1 detection and its application 
in diagnosis of lung cancer

HE Qiu-li1,ZHANG Xu2,WANG Juan2,ZHAI Rui-ping2,SUN Xia-xia2,QIAO Cai-xia1, NI Wei-hua2,GAO Su-jun1,TAI Gui-xiang2   

  1. 1.Tumor Center,First Hospital,Jilin University,Changchun 130021,China;
    2. Department of Immunology,Norman Bethune  College of Medicine,Jilin Univers
    ity,Changchun 130021,China
  • Received:2012-09-27 Online:2013-03-28 Published:2013-03-26

摘要: 目的:建立双抗体间接夹心酶联免疫吸附法(ELISA)试剂盒,检测血清黏蛋
白1(MUC1)水平,探讨其在肺癌诊断中的应用,阐明血清MUC1蛋白检测的临床应用
价值。方法:在前期制备的MUC1蛋白和兔抗人MUC1多克隆抗体的基础上,建立以鼠抗人MUC1
单克隆抗体为包被抗体,兔抗人MUC1多克隆抗体为检测抗体的双抗体间接夹心ELISA方法,
并通过对MUC1蛋白标准品的检测,绘制出标准曲线。临床上收集48例肺癌患者、7例肺良性
疾病患者和20例健康人的血清,应用本研究建立的ELISA方法检测各组标本血清MUC1蛋白表
达水平,绘制ROC曲线,确定最佳cut-off值,得出双抗体间接夹心ELISA方法检测肺癌的敏
感度、特异度及约登指数。同时与临床应用的CA15-3试剂盒检测结果进行比较。结果:应用双抗体间接夹心ELISA法确定血清MUC1的临界值为1.98  μg?L-1,检测
肺癌的敏感度为62.5%,特异度为100%,约登指数为0.625 0。CA15-3试剂盒检测肺癌的敏感度
为18.75%,特异度为100%,约登指数为0.187 5。结论:建立的双抗体间接夹心ELISA
试剂盒检测肺癌有更高的敏感度和特异度,优于临床常用的CA15-3试剂盒。

关键词:  , 黏蛋白1, 抗黏蛋白1多克隆抗体, 酶联免疫吸附法, 肺肿瘤

Abstract: Objective  To establish an indirect sandwich enzyme-linked immunesorbent assay (ELISA) kit for detecting serum level of mucin 1(MUC1) protein and to
explore its application in diagnosis of lung cancer,and to reveal the value of MUC1 protein  detection in clinic.Methods With rabbit anti-human MUC1 monoclonal antibodies and rat anti-MUC1 polyclonal antibodies as coating antibodies and testing antibodies,respectively,a new ELISA method was developed in this study,and the standard curve was gotten.Then,the serum MUC1 levels in the peripheral blood of 48 cases with lung cancer,7 cases with lung benign diseases,and 20 healthy subjects were detected with indirect ELISA.The optimal cut-off value,the specificity,the sensitivity and the Youden index were determined with ROC curve analysis.At the same time,the serum levels of MUC1 detected by CA15-3 commercial kits were analyzed.
Results Based on indirect sandwich ELISA,the cut-off value for serum MUC1 was 1.98  μg?L-1,the sensitivity was 62.5%,the specific
ity was 100% and the Youden index was 0.6250.While based on the CA15-3 kit,the sensitivity was 18.75%,the specificity was 100% and the Youden index was 0.187 5.Conclusion The indirect ELISA kit is superior to the CA15-3 kit in detecting MUC1 in lung cancer .

Key words:  , mucin 1, anti-MUC1 polyclonal antibody, enzyme-linked immunesorbent assay, lung neoplasms 

中图分类号: 

  •