Radiation Research Division for Biotechnology, Korea Atomic Energy Research Institute, 1266 Shinjeong-dong, Jeongeup-si, Jeollabuk-do 580-185, Korea.
Sensors (Basel). 2011;11(7):6685-96. doi: 10.3390/s110706685. Epub 2011 Jun 27.
Globally, over 170 million people (ca. 3% of the World's population) are infected with the hepatitis C virus (HCV), which can cause serious liver diseases such as chronic hepatitis, evolving into subsequent health problems. Driven by the need to detect the presence of HCV, as an essential factor in diagnostic medicine, the monitoring of viral protein has been of great interest in developing simple and reliable HCV detection methods. Despite considerable advances in viral protein detection as an HCV disease marker, the current enzyme linked immunosorbent assay (ELISA) based detection methods using antibody treatment have several drawbacks. To overcome this bottleneck, an RNA aptamer become to be emerged as an antibody substitute in the application of biosensor for detection of viral protein. In this study, we demonstrated a streptavidin-biotin conjugation method, namely, the RNA aptamer sensor system that can quantify viral protein with detection level of 700 pg mL(-1) using a biotinylated RNA oligonucleotide on an Octet optical biosensor. Also, we showed this method can be used to screen inhibitors of viral protein rapidly and simply on a biotinylated RNA oligonucleotide biosensor. Among the inhibitors screened, (-)-Epigallocatechin gallate showed high binding inhibition effect on HCV NS5B viral protein. The proposed method can be considered a real-time monitoring method for inhibitor screening of HCV viral protein and is expected to be applicable to other types of diseases.
全球有超过 1.7 亿人（约占世界人口的 3%）感染丙型肝炎病毒（HCV），该病毒可导致慢性肝炎等严重肝脏疾病，并进一步引发健康问题。为了检测丙型肝炎病毒（HCV）的存在，因为它是诊断医学中的一个基本因素，所以监测病毒蛋白对于开发简单可靠的 HCV 检测方法具有重要意义。尽管在作为 HCV 疾病标志物的病毒蛋白检测方面取得了相当大的进展，但目前基于抗体处理的酶联免疫吸附测定（ELISA）检测方法存在几个缺点。为了克服这一瓶颈，RNA 适体作为抗体替代物在病毒蛋白生物传感器检测中的应用开始兴起。在这项研究中，我们展示了一种链霉亲和素-生物素偶联方法，即 RNA 适体传感器系统，该系统可以使用生物素化的 RNA 寡核苷酸在 Octet 光学生物传感器上定量检测 700pg/mL 的病毒蛋白。此外，我们还表明，该方法可用于在生物素化的 RNA 寡核苷酸生物传感器上快速简单地筛选病毒蛋白抑制剂。在所筛选的抑制剂中，(-)-表没食子儿茶素没食子酸酯对 HCV NS5B 病毒蛋白具有高结合抑制作用。所提出的方法可以被认为是 HCV 病毒蛋白抑制剂筛选的实时监测方法，有望适用于其他类型的疾病。
