Abstract:
The coronavirus disease-2019 (COVID-19) pandemic has raised the stakes for planetary health diagnostics. Because pandemics pose enormous burdens on biosurveillance and diagnostics, reduction of the logistical burdens of pandemics and ecological crises is essential. Moreover, the disruptive effects of catastrophic bioevents impact the supply chains in both highly populated urban centers and rural communities. One "upstream"focus of methodological innovation in biosurveillance is the footprint of Nucleic Acid Amplification Test (NAAT)-based assays. We report in this study a water-only DNA extraction, as an initial step in developing future protocols that may require few expendables, and with low environmental footprints, in terms of wet and solid laboratory waste. In the present work, boiling-hot distilled water was used as the main cell lysis agent for direct polymerase chain reactions (PCRs) on crude extracts. After evaluation (1) in blood and mouth swabs for human biomarker genotyping, and (2) in mouth swabs and plant tissue for generic bacterial or fungal detection, and using different combinations of extraction volume, mechanical assistance, and extract dilution, we found the method to be applicable in low-complexity samples, but not in high-complexity ones such as blood and plant tissue. In conclusion, this study examined the doability of a lean approach for template extraction in the case of NAAT-based diagnostics. Testing our approach with different biosamples, PCR settings, and instruments, including portable ones for COVID-19 or dispersed applications, warrant further research. Minimal resources analysis is a concept and practice, vital and timely for biosurveillance, integrative biology, and planetary health in the 21st century. © 2023 Mary Ann Liebert, Inc., publishers.
Authors:
Goudoudaki, S.
Kambouris, M.E.
Siamoglou, S.
Gioula, G.
Kantzanou, M.
Manoussopoulou, M.
Patrinos, G.P.
Manoussopoulos, Y.
Keywords:
biological marker; distilled water; DNA; hot water; DNA; water, adult; allele specific polymerase chain reaction; Article; bacterium detection; biosurveillance; blood; blood culture; chemical waste; controlled study; cytolysis; dilution; DNA extraction; environmental footprint; extract; female; fungal detection; genotyping; human; human tissue; male; middle aged; nonhuman; oral swab; planetary health; plant tissue; solid waste; volume; diagnosis; epidemiology; polymerase chain reaction; procedures, Biosurveillance; COVID-19; COVID-19 Testing; DNA; Humans; Polymerase Chain Reaction; Water
