丙肝病毒核心抗原荧光定量型生物条形码检测体系的建立与评价  

Establishment of fluorescent quantitative bio-bar codes assay (FQ-BCA) for ultrasensitive detection of hepatitis C virus core antigen and its assessment

作　　者：张立营[1] 冯玉奎[1] 梁冰 高博[3] 孙英姿[1] 苑同业 孙宇[4] 

机构地区：[1]济南军区青岛第二疗养院检验科,山东青岛266071 [2]济南军区第401医院检验科,山东青岛266071 [3]军事医学科学院野战输血研究所,北京100850 [4]解放军第305医院检验科,北京100017

出　　处：《军医进修学院学报》2011年第4期337-340,共4页Academic Journal of Pla Postgraduate Medical School

基　　金：青岛市公共领域科技支撑计划项目(09-1-1-28-nsh)

摘　　要：目的建立丙肝病毒核心抗原(HCVcAg)的荧光定量型生物条形码检测体系,并对检测体系进行方法学评价。方法制备标记有HCVcAg多抗及特异DNA链的金纳米颗粒探针(NP探针)和标记有HCVcAg单抗的磁性微球探针(MMP探针),形成MMP探针-HCVcAg-NP探针复合物,再利用去杂交将NP探针上标记的DNA链释放出来,通过荧光定量PCR方法鉴定这些释放的DNA链可确定丙肝病毒的存在。结果建立了HCVcAg的荧光定量型生物条形码检测体系,检测灵敏度可达100fg/ml,是相应的HCVcAg ELISA检测方法的104倍。结论本研究为发展高灵敏度丙肝病毒的荧光定量型生物条形码检测试剂盒奠定了基础。Objective To establish the highly sensitive assay of hepatitis C virus core antigen（HCVcAg） and to assess its methodology.Methods HCVcAg-and DNA chain-labeled nanoparticle probe（NP） and HCVcAg-labeled magnetic microparticle probe（MMP） with monoclonal antibodies were prepared to form a MMP-HCVcAg-NP sandwich complex.DNA chain was then released by dehybridization.Hepatitis C virus was identified based on the presence of the released DNA chain detected by fluorescent quantitative PCR.Results The fluorescent quantitative bio-bar codes assay（FQ-BCA） of HCVcAg was established with a sensitivity of about 100fg/ml which was 10 times higher than that of conventional ELISA assay.Conclusion This study lays a foundation for establishing the highly sensitive HCV FQ-BCA assay of HCVcAg.

关 键 词：丙型肝炎病毒 丙型肝炎病毒核心抗原 荧光定量型生物条形码检测 方法学评价研究 

分 类 号：R446[医药卫生—诊断学]