Novel High-Volume Direct Capture Column for Efficient Nucleic Acid Recovery from Wastewater

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Abstract

The 2020 SARS-CoV-2 pandemic renewed global interest in wastewater-based epidemiology (WBE) as a tool for monitoring public health. Molecular analyses of wastewater are often limited by the small volumes of wastewater that can be processed, due to column clogging, handling constraints, and processing time. Additionally, inhibitors in the complex wastewater matrix reduce the sensitivity of downstream assays such as RT-PCR and sequencing. To address these limitations, we developed a novel column by incorporating a hydrophobic pre-filtration layer and sequential glass fiber filters. This enhanced column design, PureBioX Xpurify Column, enables processing of 1.58× more wastewater (a 58% increase in throughput) while reducing RT-PCR inhibitors and maintaining compatibility with existing workflows. Despite a modest reduction in nucleic acid yield, the modified column consistently improved viral RNA detection sensitivity, including for SARS-CoV-2. This accessible, scalable upgrade strengthens the utility of direct capture methods in WBE-based public health surveillance.

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