Sensitivity, Throughput, and Cost Analysis of Concentration Methods for Multitarget Pathogen Wastewater Monitoring

Wastewater-based epidemiology is an efficient method
for monitoring
the transmission of diverse pathogens in communities. While various
concentration methods are used, most were selected to detect SARS-CoV-2
and other respiratory viruses. Research is needed to guide the method
selection for monitoring diverse pathogens in wastewater. In this
study, a head-to-head comparison of six different concentration methods
was performed, including direct extraction (with and without bead
beating), electronegative (HA) filtration, solid concentration, and
magnetic bead-based concentration (using Nanotrap particles; with
and without bead beating). Methods were assessed for sensitivity,
inhibitor removal, recovery rates, and cost, targeting 14 microorganisms
including viruses, bacteria, and fungal pathogens. Results showed
that the concentration method selection significantly impacts the
sensitivity and economic costs of the wastewater monitoring workflow.
While no single method was optimal for all targets, combining HA filtration
and solid methods in parallel for the same sample is recommended to
sensitively detect viruses, bacteria, and fungal pathogens. The magnetic
bead-based method can be automated but costs more per sample and is
less sensitive for some targets. This study provides data-driven insights
to enhance the reliability and cost-effectiveness of wastewater surveillance
systems that can support public health responses for a broad range
of diseases.