Nano Biomedical Research Centre, School of Biomedical Engineering , Shanghai Jiao Tong University , Shanghai 200030 , P. R. China.
Division of Biomedical Engineering, School of Engineering , University of Glasgow , Glasgow G12 8LT , U.K.
ACS Nano. 2018 Jul 24;12(7):7213-7219. doi: 10.1021/acsnano.8b03183. Epub 2018 Jul 5.
In order to combat the growing threat of global infectious diseases, there is a need for rapid diagnostic technologies that are sensitive and that can provide species specific information (as might be needed to direct therapy as resistant strains of microbes emerge). Here, we present a convenient, enzyme-free amplification mechanism for a rational hybridization chain reaction, which is implemented in a simple format for isothermal amplification and sensing, applied to the DNA-based diagnosis of hepatitis B virus (HBV) in 54 patients. During the cycled amplification process, DNA monomers self-assemble in an organized and controllable way only when a specific target HBV sequence is present. This mechanism is confirmed using super-resolution stochastic optical reconstruction microscopy. The enabled format is designed in a manner analogous to an enzyme-linked immunosorbent assay, generating colored products with distinct tonality and with a limit of detection of ca. five copies/reaction. This routine assay also showed excellent sensitivity (>97%) in clinical samples demonstrating the potential of this convenient, low cost, enzyme-free method for use in low resource settings.
为了应对日益严重的全球传染病威胁，我们需要灵敏且能够提供物种特异性信息的快速诊断技术（例如，当出现耐药微生物菌株时，这可能有助于指导治疗）。在这里，我们提出了一种方便的、无酶扩增机制，用于合理的杂交链式反应，该反应以简单的格式进行等温扩增和传感，应用于基于 DNA 的乙型肝炎病毒 (HBV) 的诊断在 54 名患者中。在循环扩增过程中，只有当存在特定的靶标 HBV 序列时，DNA 单体才会以有组织和可控的方式自组装。该机制通过超分辨率随机光学重建显微镜得到证实。所采用的格式类似于酶联免疫吸附测定法，产生具有独特色调的有色产物，检测限约为 5 个拷贝/反应。该常规检测在临床样本中也显示出优异的灵敏度（>97%），证明了这种方便、低成本、无酶方法在资源有限环境中的应用潜力。
