Design and micro/nanofabrication of microfluidic lab-on-a-chip biosensors

总体目标：拟开展的研究将应用纳米生物技术解决工程挑战，并为用于疾病诊断和农业植物保护的芯片实验室（lab-on-a-chip， LOC）设备提供有效解决方案。相关背景：DNA/RNA检测的金标准是定量逆转录聚合酶链反应（qRT-PCR）。然而，这种设备价格昂贵，而且不便于携带。此外，需要经过专业培训的技术人员来操作qRT-PCR仪器。我们提出了一种基于阻抗的LOC器件作为替代解决方案。LOC测试只需要少量的试剂，设备是全自动和便携式的。直接应用包括COVID-19病毒检测。流式细胞仪是一种市售的细胞计数设备。但该设备体积较大，不适合现场监控。我们提出了一种精确测量空气传播病原体的dielectrophoresis (DEP) LOC device。可能的应用包括实时监测空气传播的COVID-19病毒和对农产品有害的空气传播孢子。研究目的：目的1：发展等温扩增扩增核酸，然后应用阻抗传感读出测试结果。目标2：将电泳学与阻抗传感相结合，实现单细胞传感。核心专长：陈洁博士在阿尔伯塔大学的研究小组包括工程师和生物医学研究人员，专长于分子生物学，人工智能，微加工和表面化学。在过去的8年里，我们开发了一种基于阻抗的LOC设备，其功能与酶联免疫吸附试验（ELISA）相同。我们已经用LOC设备成功测试了SARS-CoV-2抗体。我们的设备和运行测试所需的所有必要组件的成本不到10美元，结果在一小时内报告。预期结果：我们的加拿大制造LOC设备可以检测飞摩尔浓度的核酸和单细胞水平的病原体。我们希望每年在顶级期刊上发表两到三篇文章。我们还计划在未来五年内培养博士后和博士生。我们希望申请两到三个专利，并将技术授权给加拿大公司，以确保我们的技术成功商业化并翻译给最终用户。

Broad Goals: The proposed research will apply nanobiotechnologies to address engineering challenges and provide effective solutions in lab-on-a-chip (LOC) devices for disease diagnosis and agricultural plant protection. Relevant Background: The gold standard for DNA/RNA tests is quantitative reverse transcription polymerase chain reaction (qRT-PCR). However, the equipment is costly and is not portable. Further, professionally trained technical personnel are required to operate qRT-PCR instruments. We propose an impedance-based LOC device as an alternative solution. LOC tests only need a tiny amount of reagents, and the devices are fully automated and portable.  Immediate applications include COVID-19 viral testing. Flow cytometry is a commercially available device for cell counting. However, the device is large and not suitable for onsite monitoring. We propose a dielectrophoresis (DEP) LOC device for accurately measuring airborne pathogens. The possible applications include real-time monitoring of airborne COVID-19 virus and airborne spores harmful to agricultural food products. Research Aims: Aim #1: Develop isothermal amplification to amplify nucleic acids and then apply impedance sensing to read out test results. Aim #2: Integrate dielectrophoresis with impedance-sensing to achieve single-cell sensing.      Core Expertise: Dr. Jie Chen's research group at the University of Alberta includes engineers and biomedical researchers with specialties in molecular biology, artificial intelligence, microfabrication, and surface chemistry. Over the past 8 years, we have developed an impedance-based LOC devices that can function the same as the enzyme-linked immunosorbent assay (ELISA). We have successful tested SARS-CoV-2 antibodies with our LOC device. The cost of our device and all the necessary components required to run a test is less than $10 with results being reported in one hour.   Expect Outcomes: Our  made-in-Canada LOC device can detect nucleic acids at femtomolar concentrations and detect pathogens at the single-cell level. We expect to publish two to three articles every year in top-ranking journals. We also expect to train postdoctoral fellows and Ph.D. students in the next five years. We expect to file two to three patents and license the technologies to Canadian companies to ensure successful commercialization of our technology and translation to end users.