Although many studies concerning the detection of influenza virus have been published, a paper-based, label-free electrochemical immunosensor has never been reported. Here, we present a cost-effective, handmade paper-based immunosensor for label-free electrochemical detection of influenza virus H1N1. This immunosensor was prepared by modifying paper with a spray of hydrophobic silica nanoparticles, and using stencil-printed electrodes. We used a glass vaporizer to spray the hydrophobic silica nanoparticles onto the paper, rendering it super-hydrophobic. The super-hydrophobicity, which is essential for this paper-based biosensor, was achieved via 30–40 spray coatings, corresponding to a 0.39–0.41 mg cm−2 coating of nanoparticles on the paper and yielding a water contact angle of 150° ± 1°. Stencil-printed carbon electrodes modified with single-walled carbon nanotubes and chitosan were employed to increase the sensitivity of the sensor, and the antibodies were immobilized via glutaraldehyde cross-linking. Differential pulse voltammetry was used to assess the sensitivity of the sensors at various virus concentrations, ranging from 10 to 104 PFU mL−1, and the selectivity was assessed against MS2 bacteriophages and the influenza B viruses. These immunosensors showed good linear behaviors, improved detection times (30 min), and selectivity for the H1N1 virus with a limit of detection of 113 PFU mL−1, which is sufficiently sensitive for rapid on-site diagnosis. The simple and inexpensive methodologies developed in this study have great potential to be used for the development of a low-cost and disposable immunosensor for detection of pathogenic microorganisms, especially in developing countries.
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机译:尽管已经发表了许多有关检测流感病毒的研究,但从未报道过基于纸的,无标记的电化学免疫传感器。在这里,我们提出了一种具有成本效益的,基于手工纸的免疫传感器,用于流感病毒H1N1的无标签电化学检测。通过用疏水性二氧化硅纳米粒子喷雾修饰纸,并使用模版印刷电极来制备此免疫传感器。我们使用玻璃蒸发器将疏水性二氧化硅纳米颗粒喷洒到纸张上,使其具有超疏水性。超疏水性是此纸基生物传感器必不可少的,它通过30–40喷涂层实现,相当于在纸上的0.39–0.41 mg cm −2 sup>纳米颗粒涂层,并产生了水接触角为150°±1°。用单壁碳纳米管和壳聚糖修饰的模板印刷碳电极可提高传感器的灵敏度,并通过戊二醛交联固定抗体。使用差分脉冲伏安法评估传感器在10到10 4 sup> PFU mL -1 sup>范围内的各种病毒浓度下的灵敏度,并评估其对MS2噬菌体的选择性和乙型流感病毒。这些免疫传感器表现出良好的线性行为,改进的检测时间(30分钟)以及对H1N1病毒的选择性,检测限为113 PFU mL -1 sup>,对现场快速诊断足够敏感。这项研究中开发的简单而廉价的方法学,特别是在发展中国家,具有巨大的潜力可用于开发低成本和一次性的用于检测病原微生物的免疫传感器。
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