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
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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。Abstract: 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.
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