TY - JOUR
T1 - Comparison of five polyethylene glycol precipitation procedures for the RT-qPCR based recovery of murine hepatitis virus, bacteriophage phi6, and pepper mild mottle virus as a surrogate for SARS-CoV-2 from wastewater
AU - Torii, Shotaro
AU - Oishi, Wakana
AU - Zhu, Yifan
AU - Thakali, Ocean
AU - Malla, Bikash
AU - Yu, Zaizhi
AU - Zhao, Bo
AU - Arakawa, Chisato
AU - Kitajima, Masaaki
AU - Hata, Akihiko
AU - Ihara, Masaru
AU - Kyuwa, Shigeru
AU - Sano, Daisuke
AU - Haramoto, Eiji
AU - Katayama, Hiroyuki
N1 - Funding Information:
This work was supported by JSPS KAKENHI Grant Numbers JP20J10268 and JP20H00259 , JST J-RAPID Grant Number JPMJJR2001 , the JST-Mirai Program Grant Numbers JPMJMI18DB , and JPMJMI18DA , GAP Fund Program of Kyoto University , and the “Startup Research Program for Post-Corona Society” of Academic Strategy Office, School of Engineering, the University of Tokyo . We really appreciate kind cooperation of Japan Society on Water Environment COVID-19 taskforce and thank Enago for the English language review. Graphical abstract was created with Biorender.com .
Publisher Copyright:
© 2021 The Authors
PY - 2022/2/10
Y1 - 2022/2/10
N2 - Polyethylene glycol (PEG) precipitation is one of the conventional methods for virus concentration. This technique has been used to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA in wastewater. The procedures and seeded surrogate viruses were different among implementers; thus, the reported whole process recovery efficiencies considerably varied among studies. The present study compared five PEG precipitation procedures, with different operational parameters, for the RT-qPCR-based whole process recovery efficiency of murine hepatitis virus (MHV), bacteriophage phi6, and pepper mild mottle virus (PMMoV), and molecular process recovery efficiency of murine norovirus using 34 raw wastewater samples collected in Japan. The five procedures yielded significantly different whole process recovery efficiency of MHV (0.070%–2.6%) and phi6 (0.071%–0.51%). The observed concentration of indigenous PMMoV ranged from 8.9 to 9.7 log (8.2 × 108 to 5.6 × 109) copies/L. Interestingly, PEG precipitation with 2-h incubation outperformed that with overnight incubation partially due to the difference in molecular process recovery efficiency. The recovery load of MHV exhibited a positive correlation (r = 0.70) with that of PMMoV, suggesting that PMMoV is the potential indicator of the recovery efficiency of SARS-CoV-2. In addition, we reviewed 13 published studies and found considerable variability between different studies in the whole process recovery efficiency of enveloped viruses by PEG precipitation. This was due to the differences in operational parameters and surrogate viruses as well as the differences in wastewater quality and bias in the measurement of the seeded load of surrogate viruses, resulting from the use of different analytes and RNA extraction methods. Overall, the operational parameters (e.g., incubation time and pretreatment) should be optimized for PEG precipitation. Co-quantification of PMMoV may allow for the normalization of SARS-CoV-2 RNA concentration by correcting for the differences in whole process recovery efficiency and fecal load among samples.
AB - Polyethylene glycol (PEG) precipitation is one of the conventional methods for virus concentration. This technique has been used to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA in wastewater. The procedures and seeded surrogate viruses were different among implementers; thus, the reported whole process recovery efficiencies considerably varied among studies. The present study compared five PEG precipitation procedures, with different operational parameters, for the RT-qPCR-based whole process recovery efficiency of murine hepatitis virus (MHV), bacteriophage phi6, and pepper mild mottle virus (PMMoV), and molecular process recovery efficiency of murine norovirus using 34 raw wastewater samples collected in Japan. The five procedures yielded significantly different whole process recovery efficiency of MHV (0.070%–2.6%) and phi6 (0.071%–0.51%). The observed concentration of indigenous PMMoV ranged from 8.9 to 9.7 log (8.2 × 108 to 5.6 × 109) copies/L. Interestingly, PEG precipitation with 2-h incubation outperformed that with overnight incubation partially due to the difference in molecular process recovery efficiency. The recovery load of MHV exhibited a positive correlation (r = 0.70) with that of PMMoV, suggesting that PMMoV is the potential indicator of the recovery efficiency of SARS-CoV-2. In addition, we reviewed 13 published studies and found considerable variability between different studies in the whole process recovery efficiency of enveloped viruses by PEG precipitation. This was due to the differences in operational parameters and surrogate viruses as well as the differences in wastewater quality and bias in the measurement of the seeded load of surrogate viruses, resulting from the use of different analytes and RNA extraction methods. Overall, the operational parameters (e.g., incubation time and pretreatment) should be optimized for PEG precipitation. Co-quantification of PMMoV may allow for the normalization of SARS-CoV-2 RNA concentration by correcting for the differences in whole process recovery efficiency and fecal load among samples.
KW - Polyethylene glycol precipitation
KW - SARS-CoV-2
KW - Surrogates
KW - Virus concentration
KW - Wastewater-based epidemiology
UR - http://www.scopus.com/inward/record.url?scp=85117135850&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85117135850&partnerID=8YFLogxK
U2 - 10.1016/j.scitotenv.2021.150722
DO - 10.1016/j.scitotenv.2021.150722
M3 - Article
C2 - 34610400
AN - SCOPUS:85117135850
SN - 0048-9697
VL - 807
JO - Science of the Total Environment
JF - Science of the Total Environment
M1 - 150722
ER -
