Application of Hydrogel Nanoparticles for Detection of Dengue Virus
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Lee, Ji-Hyun
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Abstract
Dengue infection is one of the most important mosquito-borne viral diseases whose origins can be tracked to hundreds of years ago. Over the years, the infection numbers and range have spread widely with significant amplification in the number of incidents and epidemics occurring. Factors, such as misdiagnosis, have been a contributing problem in our ability to control the incidence of disease across the world. Although there are diagnostic methodologies to detect dengue infection, many existing methods are not sensitive enough to detect early infections, making it possible to have many false-negatives. With no FDA approved vaccines or therapeutic options for treatment of dengue infection related disease manifestations including Dengue Hemorrhagic Fever (DHF) and Dengue Shock Syndrome (DSS), there has been increasing interest to find a sensitive assay to detect and quantify virus, especially at the early stages of infection. As viral titers at the early stages of infection is too low to diagnose with current diagnostic methodologies, we hypothesize that Nanotrap sample preparation technology will enable enhanced detection at the lower limits of the virus using RT-qPCR. Nanotrap technology are hydrogel particles that have been engineered and tailored with specific size-sieving shell and chemical dye baits. In this study, four types of Nanotrap particles were screened to identify a particle architecture that best enriches DENV-2: CN1030, CN2010, CN2080, and CN400. CN1030 has a reactive red bait without a shell, CN2010 has a cibacron blue bait with a shell, CN1080 has an acrylic acid bait with a shell, and CN400 is a 1:1:1 ratio of CN1030, CN2010, and CN1080. Out of these Nanotrap particles, CN1030 have been found to provide the best enrichment of DENV-2 from our initial Nanotrap particles screening experiment yielding with a 10-fold enrichment. Using CN1030, we further tested limit of detection range and compared the detection range of the DENV-2 with and without CN1030. We found in absence of CN1030, DENV-2 was detected at 1.00E1 PFU/mL, but with CN1030 DENV-2 was detected at 1.00E-1 PFU/mL; this shows a 100-fold enrichment in detection. Ongoing studies are evaluating these particles for their ability to capture and enrich the other serotypes of DENV, namely DENV-1, -3, and -4 from urine. In addition, the ability of these particles to stabilize and preserve virus is also being evaluated, using DENV-2 as a prototype.
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This thesis has been embargoed for 6 months and will not be available until June 2018 at the earliest.
Keywords
Dengue virus, CN1030, Enhanced detection, Nanotrap particles