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Wastewater-based surveillance is emerging as an important tool for the COVID-19 pandemic trending. Current methods of wastewater collection, such as grab and auto-composite sampling, have drawbacks that impede effective surveillance, especially from small catchments with limited accessibility. Passive samplers, which are more cost-effective and require fewer resources to process, are promising candidates for monitoring wastewater for SARS-CoV-2. Here, we compared traditional auto sampling with passive sampling for SARS-CoV-2 detection in wastewater. A torpedo-style 3D-printed passive sampler device containing both cotton swabs and electronegative filter membranes was used. Between April and June 2021, fifteen passive samplers were placed at a local hospital’s wastewater outflow alongside an autosampler. Reverse transcription and quantitative polymerase chain reaction (RT-qPCR) was used to detect SARS-CoV-2 in the samples after processing and RNA extraction. The swab and membrane of the passive sampler showed similar detection rates and cycle threshold (Ct) values for SARS-CoV-2 RNA for the N1 and N2 gene targets. The passive method performed as well as the grab/auto sampling, with no significant differences between N1 and N2 Ct values. There were discrepant results on two days with negative grab/auto samples and positive passive samples, which might be related to the longer duration of passive sampling in the study. Overall, the passive sampler was rapid, reliable, and cost-effective, and could be used as an alternative sampling method for the detection of SARS-CoV-2 in wastewater.

Mounting evidence suggests that solids are a reliable matrix for SARS-CoV-2 detection in wastewater, yet studies comparing solids-based methods and common concentration methods using the liquid fraction remain limited. In this study, we developed and optimized a method for SARS-CoV-2 detection in wastewater using moderate-speed centrifuged solids and evaluated it against an ultrafiltration reference method. SARS-CoV-2 was quantified in samples from 12 wastewater treatment plants from Alberta, Canada, using RT-qPCR targeting the N2 and E genes. PCR inhibition was examined by spiking salmon DNA. The effects of using different amounts of solids, adjusting the sample pH to 9.6–10, and modifying the elution volume at the final step of RNA extraction were evaluated. SARS-CoV-2 detection rate in solids from 20 mL of wastewater showed no statistically significant difference compared to the ultrafiltration method (97/139 versus 90/139, p = 0.26, McNemar’s mid-p test). The optimized wastewater solids-based method had a significantly lower rate of samples with PCR inhibition versus ultrafiltration (3% versus 9.5%, p = 0.014, Chi-square test). Our optimized moderate-speed centrifuged solids-based method had similar sensitivity when compared to the ultrafiltration reference method but had the added advantages of lower costs, fewer processing steps, and a shorter turnaround time.

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