Development of Highly Sensitive Goggle for Fluorescence-Based Detection of Middle East Respiratory Syndrome Coronavirus (MERS CoV) and Other Pathogens
[Technical Report, Final Report]
59th MDWIOWA UNIV AMES
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In this project, a fluorescence-sensitive goggle prototype was developed at the University of Akron with potential field applicability in detection and identification of viral and bacterial pathogens. The goggle prototype is a wearable device that has two lenses for capturing 3D stereoscopic images. It also has video-recording capability. Further, an especially important feature of the goggle is that it can transmit the information back to a computer for detailed analysis. At the Center for Molecular Detection CAMD, our work focused on testing and evaluation of the goggles fluorescence detection capability. To do that, we devised an in vitro fluorescence-linked immunosorbent assay FLISA. For this assay, we used purified spike protein of Middle East Respiratory Syndrome Coronavirus MERS-CoV and several anti-spike protein polyclonal and monoclonal antibodies. The fluorescent dye Alexa Fluorregistered trademark 647 conjugated to secondary antibodies was used as the fluorophore. For comparison, the fluorescence signals were also read with a BioTek plate reader. The goggle prototype was able to detect fluorescence signals with detection limit of 625 ngmL of the Alexa Fluorregistered trademark 647 goat anti-rabbit IgG in a 96-well plate format. For the detection of purified MERS-CoV spike protein, samples with the spike protein can be distinguished from samples without the spike protein. However, the fluorescence detection limit was significantly lower in comparison to the fluorescence detection limit of the BioTek plate reader in 96-well plate format. The software for the goggle prototype was easy to use and offered several options to improve the quality of the capture images and the detection limit of the fluorescence signal.
- Medicine and Medical Research