Rapid Nucleic Acid Concentration from Wastewater with Minimal Equipment: New Strategy for COVID-19 Surveillance
Date
2022-04
Authors
Long, Shannon
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Abstract
Background: As of 26 March 2022, COVID-19 has killed over six million people worldwide, with almost one million of those deaths being in the United States (World Health Organization, 2022). Wastewater-based epidemiology is an effective tool for monitoring COVID-19 on a community level and catching and suppressing emerging outbreaks (Betancourt et al., 2021). However, this requires SARS-CoV-2 RNA to be concentrated from wastewater for PCR testing. A rapid, inexpensive, and uncomplicated method of RNA concentration would facilitate widespread deployment of wastewater-based epidemiology for COVID-19, especially in low-resource settings. Hollow silica microspheres float in water and can be functionalized to bind target molecules on cells of interest (Weigum et al., 2016), so they could potentially be adapted to bind to RNA and carry it to the top of a sample.
Purpose: To demonstrate the capacity of hollow silica microspheres to isolate RNA from aqueous solution.
Methodology: Hollow silica microspheres were functionalized with chitosan, a nucleotide-binding molecule (Yang et al., 2017). The functionalized microspheres were then mixed with a fluorescent oligonucleotide for varying amounts of time, after which micrographs were taken. Fluorescence around the microspheres was quantified with ImageJ.
Research: Preliminary findings suggest that the functionalized microspheres can capture oligonucleotides in as little as ten minutes. Follow-up experiments are exploring nucleotide capture at sub-10-minute intervals.
Conclusion: Chitosan-functionalized hollow silica microspheres have potential as a tool for rapid RNA concentration from wastewater. Future experiments will focus on liberating nucleic acids from the microspheres for detection after capture, in addition to further optimization of the nucleic acid capture process, including reducing the time needed for capture.
Description
Keywords
epidemiology, wastewater, COVID-19, rapid Nucleic Acid, silica microspheres
Citation
Long, S. (2022). Rapid nucleic acid concentration from wastewater with minimal equipment: New strategy for COVID-19 surveillance. Poster presented at the International Research Conference for Graduate Students, San Marcos, Texas.