Development of a rapid pre-concentration protocol and a magnetic beads-based RNA extraction method for SARS-CoV-2 detection in raw municipal wastewater

Abstract

We demonstrate the application of a rapid pre-concentration protocol and a magnetic bead-based RNA extraction method for the detection of SARS-CoV-2 RNA from raw wastewater without the need for extensive sample treatment.

Operational Constraints of Detecting SARS-CoV-2 on Passive Samplers using Electronegative Filters: A Kinetic and Equilibrium Analysis

“…These locations were selected on the basis of access to sites during April 2021 to August 2021, when there was an increase in clinical cases of COVID-19 in Nova Scotia, as described by Parra-Guardado et al in 2021. 19 Further details on specific caseloads during the study period are located in Figure S1 . However, the authors do not have specific clinical information for the sewershed areas throughout the study period.…”

“…A magnetic-beads-based RNA extraction method was used for all sample analysis. 19 A 1 mL volume from each filter eluate was extracted for RNA according to the manufacturer’s instructions. The final volume of eluted RNA from the extraction process was 50 μL, which was subsequently used for RT-qPCR analysis.…”

“…Additional information about the sampling conditions is presented in Table S1. These locations were selected on the basis of access to sites during April 2021 to August 2021, when there was an increase in clinical cases of COVID-19 in Nova Scotia, as described by Parra-Guardado et al in 2021 . Further details on specific caseloads during the study period are located in Figure S1.…”

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

“…Increased use of WBE monitoring programs as reliable strategies for noninvasive testing of communities for SARS-CoV-2 has led to the emergence of protocols and practices that are under current validation. − The University of Miami (UM), located in Miami-Dade County, Florida, USA, implemented a WBE program during the Fall 2020 semester dedicated to monitoring SARS-CoV-2 within the wastewater of UM’s three campuses . As an ongoing collaborative effort between three shared resources/laboratories of UM, the Biospecimen Shared Resource (BSSR), the Center for AIDS Research (CFAR), and the Oncogenomics Shared Resource (OGSR), a 5-week study during the summer of 2021 was conducted to compare two sample concentration methods of wastewater via electronegative filtration (ENF) and a manual magnetic bead-based concentration method (MBC), both methods adopting similar processes of currently validated workflows. ,,− The comparison was made in the CFAR laboratory using three molecular targets (N3, B2M, and OC43). The molecular assays utilized to compare the concentration methods included a novel volcano second generation-quantitative polymerase chain reaction (V2G-qPCR), developed in-house, and a standard reverse transcriptase (RT)-qPCR nucleic acid detection approach.…”

Preparation and controlled-release properties of a dual-response acidizing corrosion inhibitor

“…The attempt to open the nano valves pores was carried out under alkaline conditions, which deprotonated the ferrocene carboxylate acid group, exulting in the repulsion between carboxylate anions and b-electrostatic and then the release of dyes. [17][18][19][20] As an important additive for metal corrosion protection, corrosion inhibitor has the characteristics of high volatility and high price. In the process of oil well acidication, it is also easy to lose and lose efficacy, resulting in huge waste.…”