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. 2022 Nov 20:848:157613.
doi: 10.1016/j.scitotenv.2022.157613. Epub 2022 Jul 25.

Development of a magnetic nanoparticle-based method for concentrating SARS-CoV-2 in wastewater

Made Sandhyana Angga  1 , Bikash Malla  2 , Sunayana Raya  1 , Ayame Kitano  3 , Xiaomao Xie  4 , Hiroshi Saitoh  5 , Noriyuki Ohnishi  6 , Eiji Haramoto  7
Affiliations

Development of a magnetic nanoparticle-based method for concentrating SARS-CoV-2 in wastewater

Made Sandhyana Angga et al. Sci Total Environ. .

Abstract

Several virus concentration methods have been developed to increase the detection sensitivity of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in wastewater, as part of applying wastewater-based epidemiology. Polyethylene glycol (PEG) precipitation method, a method widely used for concentrating viruses in wastewater, has some limitations, such as long processing time. In this study, Pegcision, a PEG-based method using magnetic nanoparticles (MNPs), was applied to detect SARS-CoV-2 in wastewater, with several modifications to increase its sensitivity and throughput. An enveloped virus surrogate, Pseudomonas phage φ6, and a non-enveloped virus surrogate, coliphage MS2, were seeded into wastewater samples and quantified using reverse transcription-quantitative polymerase chain reaction to assess the recovery performance of the Pegcision. Neither increasing MNP concentration nor reducing the reaction time to 10 min affected the recovery, while adding polyacrylic acid as a polyanion improved the detection sensitivity. The performance of the Pegcision was further compared to that of the PEG precipitation method based on the detection of SARS-CoV-2 and surrogate viruses, including indigenous pepper mild mottle virus (PMMoV), in wastewater samples (n = 27). The Pegcision showed recovery of 14.1 ± 6.3 % and 1.4 ± 1.0 % for φ6 and MS2, respectively, while the PEG precipitation method showed recovery of 20.4 ± 20.2 % and 18.4 ± 21.9 % (n = 27 each). Additionally, comparable PMMoV concentrations were observed between the Pegcision (7.9 ± 0.3 log copies/L) and PEG precipitation methods (8.0 ± 0.2 log copies/L) (P > 0.05) (n = 27). SARS-CoV-2 RNA was successfully detected in 11 (41 %) each of 27 wastewater samples using the Pegcision and PEG precipitation methods. The Pegcision showed comparable performance with the PEG precipitation method for SARS-CoV-2 RNA concentration, suggesting its applicability as a virus concentration method.

Keywords: Magnetic nanoparticle; Pegcision; Polyethylene glycol precipitation; SARS-CoV-2; Virus concentration; Wastewater-based epidemiology.

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Conflict of interest statement

Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Ayame Kitano, Xiaomao Xie, Hiroshi Saitoh, and Noriyuki Ohnishi are employees of the JNC Corporation.

Figures

Unlabelled Image
Graphical abstract
Fig. 1
Fig. 1
Concentration of viruses in liquid portion per timeframe and total recovery in solid portion.
Fig. 2
Fig. 2
Detection of SARS-CoV-2 RNA in wastewater samples (n = 18) and its comparison with clinical data (N.D., not detected).
Fig. 3
Fig. 3
Recovery of φ6 and MS2, and concentration of PMMoV (log copies/L) (n = 27 each) in wastewater samples using the Pegcision and PEG precipitation methods.
See this image and copyright information in PMC

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Supplementary concepts