Aptasensor development for detection of virus in water

Julie Kirkegaard

    Research output: Book/ReportPh.D. thesisResearch

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    Contamination of water by waterborne viruses causes serious health issues worldwide. The current virus detection methods are expensive and time-consuming and require access to well-equipped laboratories. This thesis describes the development of an impedimetric all-polymer aptasensor for detection of three types of waterborne viruses: norovirus, rotavirus and hepatitis A virus. The development of the aptasensor involved sample preparation for aptamer selection of rotavirus and hepatitis A virus, an iterative design process of the aptasensor, investigation of the surface immobilisation of aptamers and finally an impedimetric electrical characterisation of the sensor.
    The sample preparation of the rotavirus was based on purification and biotinylation of the virus to meet the requirements of the aptamer selection process. The selection process, performed by an external collaborator, was based on streptavidin coated magnetic bead separation, hence the needed biotinylation. It was found that the BPH linker gave the highest yield when the biotinylated rotavirus were immobilised onto the beads.
    The design of the viral aptasensor was determined by an iterative design process. The final chip design was based on a SD card design with an injection moulded PC substrate and lid. The electrodes were screen-printed PEDOT:PSS.
    The surface immobilisation of aptamers through UV cross-linking onto different polymer substrates was tested. As the success of this step is crucial for the aptasensor specificity and performances, the surface immobilisation was throughly investigated. The aptamer UV cross-linking onto PEDOT:PSS was promising. Furthermore, some passive absorption of the aptamers onto the PEDOT:PSS was found.
    The impedimetric electrical characterisation of the aptasensor chip was done with different media salinity and different pH values. The impedimetric measurements of the different media salinity showed the expected behaviour with the greatest change present in the region representing the solution resistance. The pH measurements did not show any significant change of the impedance, hence the chip was stable in the measured pH range, which corresponds to the expected pH range of water samples.. The stability of the aptasensor chip was tested over a 2 week period in continuous flow. It was found that the electrodes were not damaged or degraded during the time period, as a constant impedance signal was measured.
    A solid foundation for the further development of the aptasensor for viral detection has been established and from this a new cheap and simple viral detection method can emerge.
    Original languageEnglish
    PublisherDTU Nanotech
    Number of pages214
    Publication statusPublished - 2016


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