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Simple SARS-CoV-2 concentration methods for wastewater surveillance in low resource settings - PubMed

  • ️Mon Jan 01 2024

Simple SARS-CoV-2 concentration methods for wastewater surveillance in low resource settings

Mohammad Dehghan Banadaki et al. Sci Total Environ. 2024.

Abstract

Wastewater-based epidemiology (WBE) measures pathogens in wastewater to monitor infectious disease prevalence in communities. Due to the high dilution of pathogens in sewage, a concentration method is often required to achieve reliable biomarker signals. However, most of the current concentration methods rely on expensive equipment and labor-intensive processes, which limits the application of WBE in low-resource settings. Here, we compared the performance of four inexpensive and simple concentration methods to detect SARS-CoV-2 in wastewater samples: Solid Fraction, Porcine Gastric Mucin-conjugated Magnetic Beads, Calcium Flocculation-Citrate Dissolution (CFCD), and Nanotrap® Magnetic Beads (NMBs). The NMBs and CFCD methods yielded the highest concentration performance for SARS-CoV-2 (∼16-fold concentration and ∼ 41 % recovery) and require <45 min processing time. CFCD has a relatively low consumable cost (<$2 per four sample replicates). All methods can be performed with basic laboratory equipment and minimal electricity usage which enables further application of WBE in remote areas and low resource settings.

Keywords: Infectious diseases; Low-resource setting; SARS-CoV-2; Viral concentration; Wastewater-based epidemiology.

Copyright © 2023 Elsevier B.V. All rights reserved.

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Conflict of interest statement

Declaration of competing interest Scott Berry has an ownership interest in Salus Discovery, LLC, which has licensed the ESP technology described in the text. Dr. Berry has also been granted patents related to the ESP process.

Figures

Figure 1.
Figure 1.

Schematic of the concentration methods. Split wastewater samples from WWTPs were processed using different concentration methods followed by ESP extraction and the extracted viral RNA was quantified by RT-qPCR.

Figure 2.
Figure 2.

Schematic of exclusion-based sample preparation (ESP). The beads are collected on a hydrophobic strip using the head magnet. With a slide of the head, the beads are positioned on the next well (wash or elution buffer). The beads are dropped in the well using the base magnet.

Figure 3
Figure 3

SARS-CoV-2 copies in the final PCR reaction from a) DES, b) SF, c) PGM-MB, d) CFCD, e) NMB vs direct extraction from wastewater (DEW). Each data point represents a unique wastewater sample, with x and y-axis values showing the DEW and the corresponding concentration method, respectively. Dashed line represents the equity line. The points’ positions relative to the dashed equity line indicate the degree of SARS-CoV-2 concentration relative to the unconcentrated samples. The closer the data points are to the upper left of each graph (the darker shade), the higher the concentration efficiency.

Figure 4
Figure 4

Performance comparison of each method a) SARS-CoV-2 concentration compared to the DEW method b) CrAssphage fold concentration c) SARS-CoV-2 recovery efficiency of each method d) CrAssphage recovery efficiency. The dashed line in a and b represents the average fold concentration. Box plots show the range, median, and average (showed by x) of recovery efficiency.

Figure 5
Figure 5

SARS-CoV-2 viral load trend in wastewater samples from a) WWTP B and b) WWTP C using different concentration methods.

Figure 6
Figure 6

Time, cost, and performance comparison of different concentration methods.

Figure 7
Figure 7

SARS-CoV-2 copies per reaction for a) NMBs and b) PGM-MBs concentration methods with and without initial centrifugation.

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