Huang Rongrong, Xi Zhijiang, Deng Yan, He Nongyue
State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China.
Medical School of Yangtze University, Jingzhou 434023, China.
Sci Rep. 2016 Aug 8;6:31103. doi: 10.1038/srep31103.
This research is aimed at selecting specific aptamer of hepatitis B e antigen by SELEX and its applications. Hepatitis B e antigen (HBeAg) seroconversion is used as an indicator of virological response when treating patients suffering from chronic hepatitis B. HBeAg also indicates a high viremia and high infectivity in untreated patients. With HBeAg modified magnetic beads as targets, three groups of aptamers are successfully selected. These are the first reported DNA aptamers that can specifically bind to HBeAg. Based on the property that the conformation changes upon binding to its target, aptamer has emerged as ideal candidate in a variety of sensing applications. In this study, we present a simple strategy for aptamer-based fluorescence biosensors for the quantitative detection of HBeAg, in which a fluorescence labeled HBeAg aptamer serves as the molecular recognition element and a short DNA molecule that is complementary to the aptamer serves as the competitor. The LOD for HBeAg is 609 ng/mL. Later, the fluorescence system is deployed in HBeAg positive and negative blood serum (p < 0.05). The total detection assay could be completed in 2 min. These newly isolated aptamers could assist the diagnosis of chronic hepatitis B.
本研究旨在通过指数富集的配体系统进化技术（SELEX）筛选乙型肝炎e抗原（HBeAg）的特异性适配体及其应用。在治疗慢性乙型肝炎患者时，HBeAg血清学转换被用作病毒学应答的指标。HBeAg还表明未治疗患者存在高病毒血症和高传染性。以修饰有HBeAg的磁珠为靶标，成功筛选出三组适配体。这些是首次报道的能特异性结合HBeAg的DNA适配体。基于与靶标结合时构象会发生变化的特性，适配体已成为各种传感应用中的理想候选物。在本研究中，我们提出了一种基于适配体的荧光生物传感器用于定量检测HBeAg的简单策略，其中荧光标记的HBeAg适配体作为分子识别元件，与该适配体互补的短DNA分子作为竞争剂。HBeAg的检测限为609 ng/mL。随后，将该荧光系统应用于HBeAg阳性和阴性血清检测（p < 0.05）。整个检测过程可在2分钟内完成。这些新分离的适配体可辅助慢性乙型肝炎的诊断。
