Viral Safety Laboratory of the National Center of Biomedical Analysis, Institute of Transfusion Medicine, The Academy of Military Medical Sciences, Beijing 100850, China.
J Virol Methods. 2012 Jul;183(1):45-8. doi: 10.1016/j.jviromet.2012.03.027. Epub 2012 Mar 30.
A simple gold nanoparticle (GNP) probe based assay (GNPA) that was modified from a bio-barcode assay (BCA) technique, was developed for ultra-sensitive, rapid detection of the bluetongue virus (BTV) VP7 outer-core protein. This assay captures the VP7 target antigen using the GNP probe coated with anti-VP7 polyclonal antibodies and single-stranded signal DNA. Magnetic microparticle (MMP) probes coated with anti-VP7 monoclonal antibodies were then added to form a sandwich immuno-complex. The single-stranded signal DNA coated onto the GNP probe present in the immuno-complex could then be detected by PCR and real-time fluorescence PCR using a TaqMan probe. The assay has a purified VP7 detection limit of 10(-2)fg/ml which is 8 orders of magnitude greater than that of conventional antigen capture ELISAs and 1 order of magnitude more sensitive than that of a conventional BCA. These results indicate that the GNPA is a highly sensitive method for easy detection of BTV proteins and that it can be modified as needed to measure the presence of other proteins.
一种基于简单金纳米粒子（GNP）探针的测定法（GNPA），是从生物条码测定法（BCA）技术改良而来，被开发用于超灵敏、快速检测蓝舌病毒（BTV）VP7 外壳蛋白。该测定法使用包被有抗-VP7 多克隆抗体的 GNP 探针捕获 VP7 靶抗原，以及单链信号 DNA。然后添加包被有抗-VP7 单克隆抗体的磁性微粒（MMP）探针，形成夹心免疫复合物。存在于免疫复合物中的单链信号 DNA 包被的 GNP 探针可以通过 PCR 和使用 TaqMan 探针的实时荧光 PCR 进行检测。该测定法对纯化的 VP7 的检测限为 10(-2)fg/ml，比传统的抗原捕获 ELISA 高 8 个数量级，比传统的 BCA 灵敏 1 个数量级。这些结果表明，GNPA 是一种用于检测 BTV 蛋白的高灵敏度方法，并且可以根据需要进行修改以测量其他蛋白的存在。
