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Abstract
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.
Highlights
There has been growing demand for low-cost, point-of-care biosensors that require minimal instrumentation, especially in developing countries
We present a low-cost paper-based immunosensor for the label-free electrochemical detection of influenza virus H1N1 using paper modified with hydrophobic silica nanoparticles (NPs) and stencil-printed electrodes
Measurements), and (iv) sensitivity, selectivity, and stability studies of the immunosensor for detection of H1N1 viruses are discussed
Summary
There has been growing demand for low-cost, point-of-care biosensors that require minimal instrumentation, especially in developing countries. Paper-based biosensors are one such class that fulfill these requirements [1]. Paper is a popular substrate material for biosensors because of its excellent characteristics, for example, its mechanical properties such as flexibility, high specific stiffness, and lightness. Most paper-based biosensors can be categorized into optical and electrochemical sensors. Colorimetric detection, such as in a home pregnancy test kit, is widely employed for many paper-based sensors, but only semi-quantification can be achieved with this technique. A great deal of research has been directed toward the development of sensitive paper-based biosensors that can provide quantitative information as well, by using surface-enhanced Raman spectroscopy, fluorescence spectroscopy, etc. A great deal of research has been directed toward the development of sensitive paper-based biosensors that can provide quantitative information as well, by using surface-enhanced Raman spectroscopy, fluorescence spectroscopy, etc. [3,4]
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