Magnetic bead technology for viral RNA extraction from serum in blood bank screening

Albertoni, Guilherme; Arnoni, Carine Prisco; Araújo, Pedro Henrique de; Andrade, Sheila Siqueira; Carvalho, Felipe Gonçalves de; Girão, Manoel Joâo Batista Castello; Schor, Nestor; Barreto, José Augusto

doi:10.1590/s1413-86702011000600008

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References 22 publications

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“…In the last decade, several studies have demonstrated the use of magnetic beads to extract nucleic acids from a variety of matrices, including serum (Sun et al, 2014), urine (Shan et al, 2012), sputum (He et al, 2017), whole blood (Albertoni et al, 2011), tissue (Mathot et al, 2011), and wastewater (La Rosa et al, 2010; Yuan et al, 2019). While conventional approaches involve both pre-concentration and extraction using magnetic beads, an emerging methodology involves direct extraction using magnetic beads in the absence of a pre-concentration step.…”

Section: Introductionmentioning

confidence: 99%

Development and optimization of a new method for direct extraction of SARS-CoV-2 RNA from municipal wastewater using magnetic beads

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Sweeney

Hayes

et al. 2020

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The use of magnetic beads in the extraction of nucleic acids from wastewater is presented as an approach to simplify extraction techniques for the detection of SARS-CoV-2 viral fragments in wastewater. In particular, this paper describes the development and optimization of a direct method for extracting SARS-CoV-2 RNA from municipal wastewater using magnetic beads. The recovery efficiency of the method using Accuplex SARS-CoV-2 Positive Reference Material (ASCV-2) was examined. Method factors assessed were sample volume, concentration of magnetic bead mix, elution temperature, and water matrix (deionized (DI) water and wastewater). The combination of optimized method parameters that resulted in the highest RNA recovery in both DI water (26.0 ± 0.8%) and wastewater (11.8 ± 1.4%) was a sample volume of 1.0 mL, a magnetic beads concentration of 100 µL mL-1 sample, and an elution temperature of 60 °C. The performance of this optimized method was further assessed in recovery experiments using wastewater samples spiked at 1.8×106 and 1.8×104 gene copies L-1 (GU L-1) with Gamma Inactivated SARS-COV-2 (GI-SCV-2) and 1.0×106 and 1.0×104 infectious units L-1 of Human Coronavirus 229E (HCV 229E) as viral surrogates. Recoveries of 86.1 and 4.6% were achieved for wastewater samples spiked with GI-SCV-2 at low and high concentrations, respectively. In assessing the effects of wastewater pre-filtration and addition of DL-Dithiothreitol (DTT, used to inactivate RNases that may degrade RNA) on recovery efficiency of ASCV-2, the magnetic bead-based extraction protocol performed optimally with unfiltered wastewater without DTT (recovery = 17.4 ± 0.4%). The method limit of detection (MLOD) for ASCV-2 recovered from pre-filtered wastewater was determined to be 4.6×104 GU L-1 (95% degree of confidence). Using this optimized magnetic bead-based extraction protocol, the presence of SARS-CoV-2 RNA was verified in wastewater collected from sewershed locations in Atlantic Canada. This emerging RNA extraction method is direct, rapid, and does not require the use of specialized equipment, thus offering advantageous application for laboratories with limited resources. As such, this method is an indispensable tool in the monitoring of wastewater for SARS-CoV-2 to potentially understand COVID-19 infection occurrence within communities and inform public health leaders.Graphical Abstract

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Section: Introductionmentioning

confidence: 99%

Development and optimization of a new method for direct extraction of SARS-CoV-2 RNA from municipal wastewater using magnetic beads

Guardado

Sweeney

Hayes

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

Magnetic bead technology for viral RNA extraction from serum in blood bank screening

Abstract: The manual magnetic bead-based extraction in combination with real-time PCR detection can be used to routinely screen blood donation for viremic donors to further increase the safety of blood products.

Cited by 1 publication

References 22 publications

Development and optimization of a new method for direct extraction of SARS-CoV-2 RNA from municipal wastewater using magnetic beads

Development and optimization of a new method for direct extraction of SARS-CoV-2 RNA from municipal wastewater using magnetic beads

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