ABSTRACT The COVID-19 pandemic has underscored the emergent need for efficient and scalable methods to predict viral outbreaks at the community level. Wastewater-based epidemiology (WBE) has proven effective in tracking the prevalence of COVID-19 infection, but conventional methods rely heavily on sophisticated laboratory infrastructure and trained technical personnel, limiting their accessibility in resource-constrained settings. This study presents a simplified wastewater processing approach that integrates ultrafiltration with a dipstick-based RNA extraction method to enable rapid, low-cost preparation of samples for downstream detection by reverse transcription quantitative polymerase chain reaction (RT-qPCR). Ultrafiltration with a sequential filter system effectively concentrates viral particles, while dipstick extraction provides a fast, equipment-minimal RNA purification step. Using SARS-CoV-2-spiked wastewater samples, the simplified workflow produced Ct values comparable to those obtained with conventional laboratory methods. By substantially reducing processing time, reagent cost, and dependence on specialized equipment, this approach addresses major barriers to broad WBE implementation. This makes it particularly well-suited for deployment in decentralized or resource-limited settings. Future work will focus on integrating this simplified approach with a portable PCR platform for point-of-care virus detection, facilitating timely public health interventions, and supporting global pandemic preparedness.
Sun et al. (Thu,) studied this question.