Centre for Personalized Nanomedicine, Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Australia.
Plant Genetic Engineering Laboratory, School of Agriculture and Food Sciences, The University of Queensland, Australia.
Sci Rep. 2017 Jan 17;7:38896. doi: 10.1038/srep38896.
Developing quick and sensitive molecular diagnostics for plant pathogen detection is challenging. Herein, a nanoparticle based electrochemical biosensor was developed for rapid and sensitive detection of plant pathogen DNA on disposable screen-printed carbon electrodes. This 60 min assay relied on the rapid isothermal amplification of target pathogen DNA sequences by recombinase polymerase amplification (RPA) followed by gold nanoparticle-based electrochemical assessment with differential pulse voltammetry (DPV). Our method was 10,000 times more sensitive than conventional polymerase chain reaction (PCR)/gel electrophoresis and could readily identify P. syringae infected plant samples even before the disease symptoms were visible. On the basis of the speed, sensitivity, simplicity and portability of the approach, we believe the method has potential as a rapid disease management solution for applications in agriculture diagnostics.
开发快速、灵敏的植物病原体检测分子诊断方法具有挑战性。在此,我们开发了一种基于纳米粒子的电化学生物传感器,用于在一次性丝网印刷碳电极上快速灵敏地检测植物病原体 DNA。该 60 分钟的测定法依赖于通过重组酶聚合扩增(RPA)快速等温扩增目标病原体 DNA 序列,然后通过金纳米粒子电化学评估进行差分脉冲伏安法(DPV)。我们的方法比常规聚合酶链反应(PCR)/凝胶电泳灵敏 10000 倍,甚至可以在疾病症状出现之前,轻易识别出感染丁香假单胞菌的植物样本。基于该方法的速度、灵敏度、简便性和便携性,我们相信该方法有可能成为农业诊断应用中快速疾病管理的解决方案。