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Comparison of Electronegative Filtration to Magnetic Bead-Based Concentration and V2G-qPCR to RT-qPCR for Quantifying Viral SARS-CoV‐2 RNA from Wastewater

ABSTRACT: Methods of wastewater concentration (electronegative filtration (ENF) versus magnetic bead-based concentration (MBC)) were compared for the analysis of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), beta-2 micro-globulin, and human coronavirus OC43. Using ENF as the concentration method, two quantitative polymerase chain reaction (qPCR) analytical methods were also compared: volcano second generation (V2G)-qPCR and reverse transcriptase (RT)-qPCR

measuring three different targets of the virus responsible for the COVID-19 illness (N1, modified N3, and ORF1ab). Correlations between concentration methods were strong and statistically significant for SARS-CoV-2 (r = 0.77, p < 0.001) and B2M (r = 0.77, p < 0.001). Comparison of qPCR analytical methods indicates that, on average, each method provided equivalent results with average ratios of 0.96, 0.96, and 1.02 for N3 to N1, N3 to ORF1ab, and N1 to ORF1ab and were supported by significant (p < 0.001) correlation coefficients (r = 0.67 for V2G (N3) to RT (N1), r = 0.74 for V2G (N3) to RT (ORF1ab), r = 0.81 for RT (N1) to RT (ORF1ab)). Overall results suggest that the two concentration methods and qPCR methods provide equivalent results, although variability is observed for individual measurements. Given the equivalency of results, additional advantages and disadvantages, as described in the discussion, are to be considered when choosing an appropriate method.





Comparison of Electronegative Filtration to Magnetic Bead-Based Concentration and V2G-qPCR to RT-qPCR for Quantifying Viral SARS-CoV-2 RNA from Wastewater


Kristina M. Babler, Ayaaz Amirali, Mark E. Sharkey, Sion L. Williams, Melinda M. Boone, Gabriella A. Cosculluela, Benjamin B. Currall, George S. Grills, Jennifer Laine, Christopher E. Mason, Brian D. Reding, Stephan C. Schürer, Mario Stevenson, Dušica Vidović, and Helena M. Solo-Gabriele


ACS ES&T Water 2022 2 (11), 2004-2013

DOI: 10.1021/acsestwater.2c00047


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