Table 2.
Description of analytical methods from selected literature and their contributions.
| Study | Location and sampling period | Wastewater type and sources | Sample pre-treatment | Concentration methods | RNA extraction Kit/protocol | Quantification method and gene targets | Analyzed initial sample volume (mL) | Type of contribution (Fig. 2) |
|---|---|---|---|---|---|---|---|---|
| Anderson-Coughlin et al. (2021) | USA August 2020–March 2021 |
Raw (sewage) | Filtration using a 0.22 μm polyethersulfone (PES) membrane | Centrifugal ultrafiltration | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N1 and N2 |
40 – 45 | 3 |
| Ahmed et al. (2020a) | Australia March 2020 – April 2020 |
Raw (influent) | (1) pH adjustment to 3.5/4 using HCl, (2) Centrifugation at 4750 g for 30 min | (1) Adsorption-elution with electronegative membranes, (2) Ultrafiltration | QIAgen RNeasy PowerWater Kit and QIAgen RNeasy PowerMicrobiome Kit |
RT-qPCR N |
100 – 200 | 5 |
| Ahmed et al. (2020c) | Australia NR |
Raw (influent) | (1) Acidification to pH 4 using 2 N HCl, (2) NR, (3) MgCl2 addition to a final concentration of 25 mM MgCl2, (4,5) Centrifugation at 4500 g for 10 min at 4 °C, (6) Centrifugation at 10000 g for 20 min at 4 °C, (7) Centrifugation at 10000 g for 1 h at 4 °C | (1,2,3) Adsorption-elution using electronegative membranes, (4,5) Ultrafiltration, (6) PEG precipitation, (7) Ultracentrifugation | QIAgen RNeasy PowerMicrobiome Kit | RT-qPCR NR |
50 | 1,5 |
| Ahmed et al. (2021a) | Bangladesh July 2020 – August 2020 |
Raw (sewage) | Centrifugation at 4500 g for 30 min, filtration using 0.22 μm filters | PEG precipitation | Favor Prep Viral Nucleic Acid Extraction Kit | RT-qPCR N and ORF1ab |
50 | 1,5 |
| Ahmed et al. (2021b) | NR | Raw (influent) | (1) Centrifugation at 4000 g for 30 min at 4 °C, (2) NR | (1) Concentrating pipette (InnovaPrep), (2) Adsorption-elution with electronegative membranes | QIAgen QIAamp® Viral RNA mini Kit and RNeasy PowerWater Kit | RT-qPCR N1 |
NR | 1 |
| Ahmed et al. (2021c) | Australia February 2020 – May 2020 |
Raw (influent) | NR | Adsorption-elution using electronegative membranes | QIAgen RNeasy PowerMicrobiome Kit | RT-qPCR N1, N2 and N3 |
100 – 200 | 2 |
| Ahmed et al. (2022b) | Australia June 2021 |
Raw (influent) | Centrifugation at 3000 g for 5 min | Concentration Pipette (InnovaPrep) | QIAgen QIAamp® Viral RNA mini Kit and QIAgen RNeasy PowerMicrobiome Kit (for the solid phase) | RT-qPCR and RT-dPCR N1 and N2 |
50 | 4,6 |
| Ai et al. (2021) | USA July 2020–January 2021 |
Raw (influent) | Centrifugation at 2500 g for 10 min at 4 °C, filtration using a 0.45 μM sterile filter unit | Sequential concentration using adsorption-elution with positively charged membranes, organic flocculation, and centrifugal ultrafiltration | QIAgen RNeasy PowerMicrobiome Kit | RT-ddPCR N1, N2 and E |
100 – 200 | 4,5,6 |
| Amereh et al. (2022) | Iran September 2020 – April 2021 |
Raw (influent) | Centrifugation at 4000 g for 10 min | PEG precipitation | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N and ORF1ab |
50 | 4 |
| Amoah et al. (2021) | South Africa NR |
Raw (influent) | Heat inactivation 60 °C for 90 min | Centrifugal ultrafiltration | QIAgen QIAamp® Viral RNA mini Kit | RT-ddPCR and RT-LAMP E, N, ORF1ab, RdRP and S |
250 | 3 |
| Anneser et al. (2022) | USA March 2020 – March 2021 |
Raw (influent and sludge) | NR | (1) PEG precipitation, (2) Spectrophotometry | TRIzol-chloroform protocol and RNeasy PowerSoil Total RNA Kit | RT-qPCR N1, N2 and N3 |
NR | 4.5 |
| Arora et al. (2020) | India May 2020 – June 2020 |
Raw (influent) | Heat inactivation 60 °C for 90 min, filtration using a 0.45 μm membrane | PEG precipitation | Allplex 2019-nCoV Assay Kit | RT-PCR N, S, E, ORF1ab and RdRp |
50 | 2 |
| Bagutti et al. (2022) | Switzerland July 2021–December 2021 |
Raw (influent) | NR | NR | Maxwell® RSC Environ Wastewater TNA Kit | RT-qPCR N1, N2 and E |
40 | 4 |
| Baldovin et al. (2021) | Italy April 2020 – May 2020 |
Raw (influent) and treated (effluent: activated sludge, peractic acid and UV lamps) | Filtration using a 0.22 μm PES membrane | Ultrafiltration | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N and ORF1ab |
100 | 1,6 |
| Barbosa et al. (2022) | Brazil May 2020–October 2020 |
Raw (influent, sewage) | NR | Ultracentrifugation | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N1 and N2 |
40 | 2,4,5 |
| Bar-Or et al. (2021a) | Israel August 2020–February 2021 |
Raw (influent) | Centrifugation at 4696 g for 5 min | Adsorption-elution with electronegative membranes | NucliSENS EasyMAG | RT-qPCR E |
25 | 2 |
| Bar-Or et al. (2021b) | Israel March 2020–April 2020 |
Raw (sewage) | NR | (1) PEG precipitation, (2) Skimmed milk flocculation, (3) Ultrafiltration | QIAgen RNeasy mini Kit and NucliSENS EasyMAG | RT-qPCR N and E |
250 – 1000 | 1,2,6 |
| Barril et al. (2021) | Argentina March 2020–October 2020 |
Raw (influent) | NR | A total of 11 different methods were evaluated. | Maxwell RSC 48 Extraction System | RT-qPCR N1 and N2 |
Varied from method to method | 2,4 |
| Barrios et al. (2021) | Argentina June 2020 – April 2021 |
Raw (influent) | Heat inactivation 60 °C for 90 min | PEG precipitation | TRIzol-chloroform protocol | RT-qPCR N1 |
200 | 4 |
| Barua et al. (2022) | USA June 2020 – November 2020 |
Raw (influent) | Heat inactivation 75 °C for 40 min | Electronegative filtration (HA) | QIAgen QIAamp® Viral RNA mini Kit and NucliSENS EasyMAG | RT-qPCR and RT-ddPCR N1 and N2 |
20 | 1 |
| Bertrand et al. (2021) | France April 2020–May 2020 |
Raw (influent after decantation) | NR | (1) Ultrafiltration, (2) PEG precipitation | Phenol-chloroform-isoamyl alcohol protocol | RT-PCR and RT-ddPCR E and RdRp |
50 | 1,2 |
| Bivins et al. (2022) | NR | Raw (influent, sewage) | NR | Centrifugal ultrafiltration | QIAgen QIAamp® Viral RNA mini Kit and AllPrep PowerViral DNA/RNA Kit | RT-ddPCR and RT-LAMP N2 and E |
NR | 1,3,4 |
| Boogaerts et al. (2021) | Belgium August 2020 – January 2021 |
Raw (influent) | (1) Centrifugation at 4600 g for 30 min at 4 °C, (2) Centrifugation at 4654 g for 30 min at 4 °C | (1) Ultracentrifugation, (2) PEG precipitation | QIAgen QIAamp® Viral RNA mini Kit, RNeasy plus miniKit and QIAgen RNeasy PowerMicrobiome Kit |
RT-qPCr and RT-dPCR N1, N2, N3 and E |
20 – 90 | 1,4,5 |
| Boogaerts et al. (2022) | Belgium September 2020 – November 2021 |
Raw (influent) | Centrifugation at 4000 g for 30 min | Ultracentrifugation | Maxwell® RSC PureFood GMO and Authentication Kit | RT-qPCr and RT-dPCR N, S and E |
20 | 3,6 |
| Calderon-Franco et al. (2022) | Netherlands July 2020 – December 2020 |
Raw (influent) | Heat inactivation 65 °C for 30 min | (1) Adsorption-elution with electronegative membranes, (2) Polyethersulfone membranes, (3) Anion-exchange diethylaminethyl cellulose columns | Fast RNA Blue Kit, FAST RNA Kit and MagMax CORE Nucleic Acid Purification Kit | RT-qPCR S, N and ORF1ab |
50 – 550 | 1,3 |
| Canh et al. (2021) | Japan January 2021 – February 2021 |
Raw (influent) | Centrifugation at 3500 g for 15 min | (1) Ultrafiltration, (2) PEG precipitation | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N1 |
100 | 1,2 |
| Carrillo-Reyes et al. (2021) | Mexico April 2020–July 2020 |
Raw (influent, sewage) and treated (secondary sludge, effluent) | Filtration using a 0.2 μm PES membrane | (1) Ultrafiltration, (2) Adsorption-elution with electronegative membranes | QIAgen RNeasy PowerMicrobiome Kit | RT-qPCR RdRp, S and E |
(1) 120, (2) 30 – 100 |
1,2 |
| Castiglioni et al. (2022) | March 2020–June 2020 | Raw (influent) | Under UV light for 30 min, Centrifugation at 4500 g for 30 min at 4 °C | PEG precipitation | QIAgen QIAamp® MinElute Virus Spin Kit | RT-PCR N1 and N3 |
45 | 1,2,5 |
| Chakraborty et al. (2021) | India September 2020 |
Raw (influent) and treated (primary sludge, effluent) | NR | Composite, Supernatant, Sediment and Syringe Filtration | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N1 and N2 |
250 | 1 |
| Chavarria-Miro et al. (2021) | Spain April 2020 – July 2020 |
Raw (influent) | NR | PEG precipitation | NucliSENS miniMAG | RT-qPCR N1, N2, RdRp, IP2 and IP4 |
800 | 5 |
| Claro et al. (2021) | Brazil June 2020 – April 2021 |
Raw (influent) | Centrifugation at 8000 g for 120 min at 4 °C | PEG precipitation | PureLink™ Viral RNA/DNA mini Kit | RT-qPCR N1 and N2 |
40 | 4 |
| D'Aoust et al. (2021) | Canada April 2020–June 2020 |
Raw (post-grid influence) and treated (primary clarified sludge) | Decantation and serially filtered through a 1.5 μm glass fiber filter followed by a 0.45 μm GF6 mixed cellulose-ester filter |
PEG precipitation | QIAgen RNeasy PowerMicrobiome Kit | RT-qPCR and RT-ddPCR N1 and N2 |
32 | 3,5 |
| de Freitas et al. (2022) | Brazil January 2021 – January 2022 |
Raw (influent) | NR | PEG precipitation | PureLink Viral RNA/DNA mini Kit | RT-qPCR N1 and N2 |
40 | 4.5 |
| de Sousa et al. (2022) | Brazil January 2021 – August 2021 |
Raw (influent) and treated (effluent) | pH adjustment to 3.5 using 1 M HCl, shaken at °4C for 30 min, Centrifugation at 2474 g for 30 min at 4 °C | PEG precipitation | MagMAX Viral/Pathogen Nucleic Acid Isolation Kit | RT-qPCR N1 and N2 |
50 | 5 |
| Dimitrakopoulos et al. (2022) | Greece November 2021–December 2021 |
Raw (influent) | NR | (1) PEG precipitation, (2) PEG precipitation with glycine, (3) Direct capture, (4) Adsorption-elution with electronegative membranes, (5) Ultrafiltration | Water DNA/RNA magnetic bead Kit, QIAgen RNeasy PowerMicrobiome Kit, AllPrepPowerViral DNA/RNA Kit and Manual EnviroWastewater TNA Kit | RT-qPCR and RT-ddPCR N1, N2 and N3 |
50 | 2,6 |
| Dumke et al. (2021) | Germany, NR |
Raw (influent) | Centrifugation at 3300 g for 30 min at 4 °C | (1) PEG precipitation, (2) Centrifugation with Vivaspin columns | QIAgen RNeasy kits (not specified what series) | RT-qPCR and RT-ddPCR S and E |
40 | 2 |
| Farkas et al. (2021) | NR | Raw (NR) | Centrifugation at 3000 g for 30 min at 4 °C or 1000 g for 10 min at 4 °C, pH adjustment of supernatant to 7–7.5 using 1 M NaOH | PEG precipitation | NucliSENS lysis buffer and NucliSENS miniMag extraction system | RT-qPCR N1 and N2 |
50 | 1,3 |
| Feng et al. (2021) | USA August 2020–January 2021 |
Raw (influent) | Filtration using 0.8 μm cellulose-ester filters | Bashing Bead Lysis | QIAgen RNeasy PowerMicrobiome Kit | RT-ddPCR N1 and N2 |
25 | 1 |
| Fernandez-Cassi et al. (2021) | Switzerland February 2020–April 2020 |
Raw (influent) | Filtered using 2 μm glass fiber filters | Centrifugal ultrafiltration | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N1 and N2 |
50 | 5 |
| Fitzgerald et al. (2021) | Scotland April 2020 – January 2021 |
Raw (influent) | Centrifugation at 4000 g for 30 min at 4 °C, filtration using a syringe filter | (1) Ultracentrifugation, (2) PEG precipitation, (3) skimmed milk flocculation | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N1 and E |
20 – 40 | 1,2 |
| Flood et al. (2021) | USA March 2020 – September 2020 |
Raw (influent, sewage) | (1) Centrifugation at 2500 g for 5 min at 4 °C, (2) Centrifugation at 4654 g for 30 min at 4 °C, (3) Centrifugation at 4700 g for 45 min at 4 °C | (1,2) Ultrafiltration, (3) PEG precipitation | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR, RT-ddPCR N1, N2 and E |
(1,2) 100, (3) NR | 1 |
| Fongaro et al. (2021) | Brazil October 2019–March 2020 |
Raw (sewage) | NR | PEG precipitation | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N1, S and RdRp |
25 | 4 |
| Fonseca et al. (2022) | Brazil March 2021 |
Raw (influent, river) | Heat inactivation 60 °C for 90 min, filtration using 1.2 μm pore size microfiber filters, Centrifugation at 4500 g for 30 min at 4 °C | (1) Ultrafiltration, (2) Adsorption-elution with electronegative membranes, (3) Aluminum hydroxide precipitation, (4) PEG precipitation | MagMax Viral/Pathogen II Kit and KingFisher Duo Purification System | RT-qPCR N1 and N2 |
40 | 4,6 |
| Galani et al. (2022) | Greece August 2020 – March 2021 |
Raw (influent) | Centrifugation at 4700 g for 30 min at 4 °C | (1) PEG precipitation, (2) centrifugal ultrafiltration | Water DNA/RNA Magnetic Bead Kit, QIAgen RNeasy Power Microbiome Kit and QIAgen RNeasy Serum/Plasma Advanced Kit | RT-qPCR N1 and N2 |
50 | 4,5 |
| Gerrity et al. (2021) | USA March 2020–May 2020 |
Raw (influent) | (1) NR, (2,3) Centrifugation at 3500 g for 15–30 min at 10 °C | (1) Hollow-fiber ultrafiltration, (2) Centrifugal Ultrafiltration, (3) PEG precipitation. | Purelink Viral RNA/DNA mini Kit | RT-qPCR N1, N2, E and ORF1a |
50 | 1,3 |
| Giraud-Billoud et al. (2021) | Argentina July 2020–November 2020 |
Raw (influent) | Heat inactivation 60 °C for 90 min | (1) PEG precipitation, (2) Polyaluminum chloride (PAC) flocculation | NucleoZOL | RT-qPCR N1 and N2 |
300 | 5,6 |
| Gonçalves et al. (2021) | Slovenia June 2020 |
Raw (sewage) | Filtration using a 0.70 μm glass fiber filter membrane | Ultracentrifugation | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR E and RdRp |
100 | 1 |
| Gonzalez et al. (2020) | USA March 2020 – August 2020 |
Raw (influent) | (1) Centrifugation at 10000 g for 10 min, (2) NR | (1) Concentration Pipette (InnovaPrep), (2) Adsorption-elution using electronegative membranes | NucliSENS Easy Mag TNA Extraction Kit | RT-ddPCR N1, N2 and N3 |
125 | 2 |
| Gonzalez-Reyes et al. (2021) | Mexico June 2020–July 2020 |
Raw (influent, sewage) | Heat inactivation 60 °C for 90 min, filtration using a 0.2 μm membrane | PEG precipitation | TRIzol protocol | RT-qPCR N1, N2 and N3 |
150 | 2 |
| Haramoto et al. (2020) | Japan March 2020–May 2020 |
Raw (influent) and treated (activated sludge before chlorination) | NR | (1) Adsorption-elution with electronegative membranes, (2) Direct adsorption | QIAgen QIAamp® Viral RNA mini Kit and QIAgen RNeasy PowerMicrobiome Kit | RT-qPCR N1, N2, S and ORF1ab |
200 – 5000 | 2 |
| Hasan et al. (2021) | UAE May 2020–June 2020 |
Raw (influent) and treated (effluent) | (1,2) Heat inactivation 60 °C for 90 min, filtration using a 0.22 μm PES membrane | (1) Ultrafiltration, (2) PEG precipitation | ABIOpure Viral DNA/RNA Extraction Kit and TRIzol-chloroform protocol | RT-qPCR RdRp |
(1,2) 50 | 1,4 |
| Hasing et al. (2021) | Canada October 2020–December 2020 |
Raw (influent) | pH adjustment to 9.6–10 using 5 N NaOH, Centrifugation at 4500 g for 10 min | Ultrafiltration | MagMAX96 Viral RNA Isolation Kit and King Fisher Flex Purification System | RT-qPCR N2 and E |
100 | 3 |
| Hata et al. (2021) | Japan March 2020 – April 2020 |
Raw (influent) | NR | PEG precipitation | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N2 and N3 |
80 | 2 |
| Hemalatha et al. (2021) | India July 2020 – August 2020 |
Raw (influent) | Gravity filtration with 1 mm thick blotting sheets to remove debris and larger particles followed by filtration using 0.2 μm filtration units | Centrifugal ultrafiltration | QIAamp® Viral RNA isolation Kit | RT-qPCR N, E and ORF1ab |
100 | 2,3 |
| Hoar et al. (2022) | USA April 2020 – February 2021 |
Raw (influent) | Heat inactivation 60 °C for 90 min, Centrifugation at 5000 g for 10 min at 4 °C, filtration using 0.22 μm acetate-cellulose membrane | PEG precipitation | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N1 |
40 | 4.5 |
| Hokajarvi et al. (2021) | Finland April 2020 – May 2020 |
Raw (influent) | Centrifugation at 4654 g for 30 min | Ultrafiltration | Chemagic Viral300 DNA/RNA extraction Kit | RT-qPCR N2 and E |
60 | 2,3 |
| Huang et al. (2021) | Canada October 2020 – March 2021 |
Raw (influent) | NR | Ultrafiltration | QIAgen RNeasy PowerMicrobiome Kit | RT-qPCR N1, N2, N3 and E |
200 | 2 |
| Iglesias et al. (2021) | Argentina June 2020–September 2020 |
Raw (influent, surface water) | Heat inactivation 60 °C for 90 min | PEG precipitation | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N1 and N2 |
250 | 2 |
| Jafferali et al. (2021) | Sweden and Italy May 2020–June 2020 |
Raw (influent) | (1) Centrifugation at 4600 g for 30 min at 4 °C, (2) Centrifugation at 1500 g for 15 min at 4 °C, (3) NR, (4) Centrifugation at 4600 g for 30 min at 4 °C | (1) Ultrafiltration, (2) Double Ultrafiltration, (3) Adsorption-elution with electronegative membranes, (4) Centrifugation combined with adsorption-extraction | TRIzol reagent and RNeasy PowerMicrobiome Kit | RT-qPCR N |
40 – 50 | 1 |
| Jmii et al. (2021) | Tunisia September 2020–October 2020 |
Raw (influent) | Coarse filtration and microfiltration, pH adjustment to 6 with aluminum hydroxide | Adsorption-elution with electronegative membranes | QIAgen RNeasy PowerMicrobiome Kit | RT-PCR N, E and RdRp |
100 | 1 |
| Johnson et al. (2021) | South Africa June 2020 |
Raw (influent) | Centrifugation at 3500 g for 20 min | NR | RNeasy PowerSoil Kit | RT-qPCR N1 and N2 |
50 – 100 | 2 |
| Juel et al. (2021) | USA October 2022 – March 2021 |
Raw (sewage) | NR | (1) Adsorption-elution with electronegative membranes, (2) Concentrating Pipette (InnovaPrep) | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N1 |
40 – 100 | 1,3 |
| Kevill et al. (2022) | Wales October 2020 – February 2021 |
Raw (influent) | Centrifugation at 15000 g for 10 min at 4 °C | (1) PEG precipitation, (2) Ammonium sulfate precipitation, (3) Concentration pipette (Innova Prep) | NucliSENS Lysis Buffer, NucliSENS Extraction Reagent Kit and King-Fisher 96 Flex System | RT-pPCR N1 |
200 | 1,3 |
| Kitamura et al. (2021) | Japan June 2020–August 2020 |
Raw (influent, sewage) | Centrifugation at 3000 rpm for 30 min | (1) Adsorption-elution with electronegative membranes, (2) PEG precipitation, (3) Ultrafiltration, (4) Solid precipitation | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N1 and N2 |
400 | 1,3,4,5 |
| Koureas et al. (2021) | Greece October 2020–April 2021 |
Raw (influent) | NR | PEG precipitation | MagMAX™ Viral/Pathogen Nucleic Acid Isolation Kit | RT-PCR N, S and ORF1ab |
105 | 5 |
| Krivonakova et al. (2021) | Slovakia September 2020 – March 2021 |
Raw (influent) | Centrifugation at 4700 g for 30 min | Ultracentrifugation | Direct-zol RNA miniprep Kit | RT-qPCR E, RdRp and ORF1ab |
50 | 2.4 |
| Kuhn et al. (2022) | USA November 2020 – March 2021 |
Raw (sewage) | Filtration using a 70 μm mesh cell strainer | PEG precipitation | Bio-On-Magnetic-Beads platform | RT-qPCR N1 |
32 | 5 |
| Kumar et al. (2021) | India August 2020–September 2020 |
Raw (influent) | Centrifugation at 4000 g for 40 min, filtration using a 0.22 μm syringe filter | PEG precipitation | NucleoSpin® RNA Virus isolation Kit | RT-PCR N, S and ORF1ab |
30 | 2 |
| La Rosa et al. (2020) | Italy February 2020–April 2020 |
Raw (influent) | Heat inactivation 56 °C for 30 min | PEG-dextran two-phase separation | NucliSENS miniMAG | RT-qPCR ORF1ab, S and RdRp |
250 | 2,5 |
| Langan et al. (2022) | USA January 2021–March 2021 |
Raw (sewage) | Centrifugation at 4000 g for 20 min at 4 °C | Ultrafiltration | QIAgen PowerViral DNA/RNA Kit, Zymp EnvironWater RNA Extraction Kit and Monarch Total RNA miniprep Kit | RT-qPCR N1 and N2 |
200 | 1,5 |
| Lara-Jacobo et al. (2022) | Canada October 2020–April 2021 |
Raw (influent) | Adding 50 mL of acetone at 4 °C and stored overnight at −20 °C to precipitate proteins, Centrifugation at 3405 g for 15 min | Protein Precipitation and Digestion | QIAgen PowerMicrobiome Kit | RT-qPCR N1 |
40 | 1,3 |
| LaTurner et al. (2021) | USA October 2020 |
Raw (influent) | (1) NR, (2) Centrifugation at 4100 g for 10 min at 4 °C, (3) Centrifugation at 3000 g for 1 min at 4 °C, (4) Centrifugation at 7140 g for 15 min at 4 °C, (5) Centrifugation at 4100 g for 10 min at 4 °C | (1) Direct extraction, (2) HA filtration with bead beating, (3) HA filtration with elution, (4) PEG precipitation, (5) Ultrafiltration | Chemagic Prime Viral DNA/RNA 300 Kit H96 | RT-qPCR and RT-ddPCR N1 and N2 |
(1) 1, (2,3,5) 50 (4) 200 | 5 |
| Layton et al. (2022) | USA June 2020 – July 2020 |
Raw (sewage) | Centrifugation at 12000 g for 1 min | NR | MagMAX Viral Pathogen Kit | RT-ddPCR N1 and N2 |
30 – 40 | 4 |
| Li et al. (2022) | USA June 2020 – September 2021 |
Raw (influent) | Heat inactivation 60 °C for 60 min, Centrifugation at 3000 g for 15 min, and sequential filtration using 1.5, 0.8, and 0.45 μm sterile membrane filters | PEG precipitation | AllPrep PowerViral DNA/RNA Kit | RT-qPCR N1 and N2 |
NR | 2,4 |
| Maida et al. (2022) | Italy September 2021–July 2021 |
Raw (sewage) | NR | PEG-dextran two-phase separation | NucliSENS miniMAG | RT-qPCR NR |
NR | 5 |
| Mailepessov et al. (2022) | Singapore April 2020 |
Raw (sewage) | (1) Centrifugation at 4000 g for 30 min, (2) Centrifugation at 2000 g for 5 min | (1) PEG precipitation, (2) Ultrafiltration | Modified TRIzol-QIAgen protocol and QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR NR |
45 | 1,3 |
| Markt et al. (2022) | Liechtenstein Sept 2020 - March 2021 |
Raw (influent) | Centrifugation at 4500 g for 30 min | PEG precipitation | Monarch total RNA miniprep Kit | RT-qPCR N1 |
70 | 1 |
| Masachessi et al. (2022) | Argentina May 2020–August 2021 |
Raw (influent) | Centrifugation at 4750 g for 20 min at 4 °C | PEG precipitation | MagNa Pure 96 DNA and Viral NA Large Volume Kit | RT-qPCR N and E |
500 | 2 |
| McMahan et al. (2021) | USA May 2020 – August 2020 |
Raw (sewage) | Heat inactivation 60 °C for 30 min, Centrifugation at 6500 g for 10 min at 6 °C | PEG precipitation | TRIzol-chloroform protocol | RT-qPCR N |
225 | 5 |
| McMinn et al. (2021) | USA July 2020–October 2020 |
Raw (influent, primary treated) | Heat inactivation 121 °C for 60 min | (1) Ultrafiltration, (2) Concentration Pipette | QIAgen All Prep PowerViral Kit | RT-qPCR N |
2000 | 1 |
| Mlejnkova et al. (2020) | Czech Republic April 2020–June 2020 |
Raw (influent) | NR | Skimmed milk flocculation | NucliSENS miniMAG | RT-qPCR NR |
500 | 1 |
| Mondal et al. (2021) | USA October 2020 - Jan 2021 |
Raw (influent) | NR | Direct Capture | NR | RT-qPCR N1, N2 and E |
40 | 1 |
| Monteiro et al. (2022) | Portugal April 2020–December 2020 |
Raw (influent) | Hollow-fiber filtration | PEG precipitation | QIAgen QIAamp® Fast DNA Stool mini Kit | RT-qPCR N, E and RdRp |
1000 | 1 |
| Nagarkar et al. (2022) | USA May 2020 – November 2020 |
Raw (influent) | NR | Ultrafiltration | RNeasy PowerWater Kit | dd-PCR N1 and N2 |
225 | 4 |
| Nasseri et al. (2021) | Iran April 2020–May 2020 |
Raw (influent) and treated (effluent) | Decantation for 5 min, Centrifugation at 1500 g for 20 min at 4 °C, pH adjustment to 7–7.5 using HCl and NaOH | PEG-dextran two-phase separation | FastPure Viral RNA mini Kit | RT-PCR N and ORF1ab |
250 | 2 |
| Navarro et al. (2021) | Italy December 2020 – February 2021 |
Raw (influent) | Centrifugation at 4500 g for 30 min at 4 °C | Centrifugal ultrafiltration | Quick-RNA Fecal/Soil Microbe Microprep | RT-qPCR N1, N3 and S |
100 | 1 |
| Ni et al. (2021) | Australia March 2020–April 2020 |
Raw (influent) | Centrifugation at 9000 g for 20 min at 4 °C | Ultrafiltration | QIAgen RNeasy PowerMicrobiome Kit | RT-qPCR N1 and N2 |
50 | 3,6 |
| Nourbakhsh et al. (2022) | Canada September 2020–June 2021 |
Raw (influent) | (1,2,3) Centrifugation at 4000 g for 20 min at 4 °C | (1) Centrifugal ultrafiltration, (2) Zirconia-silica beads in a Bead Mill 24 Homogenizer, (3) Centrifugation | MagNA Pure 96 DNA, Viral NA Large Volume Kit and QIAgen RNeasy PowerMicrobiome Kit | RT-qPCR N1 and N2 |
15 – 30 | 5 |
| Novoa et al. (2022) | Spain May 2020–May 2021 |
Raw (influent, sewage) and treated (effluent) | Filtration using a 20–25 μm cellulose filter, pH adjustment to 6 | Adsorption-precipitation with AlCl3 | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N1, N2 and E |
150 | 5,6 |
| O’Brien et al. (2021) | USA June 2020 |
Raw (sewage) | NR | Ultrafiltration | QIAgen All Prep PowerViral DNA/RNA KIT, Monarch RNA miniprep Kit and Zymo Quick RNA-Viral | RT-qPCR N2 |
250 | 5 |
| Parra-Guardado et al. (2022) | Canada NR |
Raw (influent) | Centrifugation at 5000 rpm for 5 min | NR | Direct Magnetic Bead Extraction | RT-qPCR NR |
50 | 1 |
| Peinado et al. (2022) | Spain February 2021 – June 2021 |
Raw (influent) | (1) Centrifugation at 4600 g for 30 min, pH adjustment to 6, (2) Centrifugation at 8000 g for 30 min at 4 °C, (3) Centrifugation at 4600 g for 30 min | (1) Adsorption-precipitation with aluminum hydroxyde, (2) PEG precipitation, (3) Ultrafiltration | NZY Viral RNA Isolation Kit | RT-qPCR N1 and N2 |
100 – 200 | 1 |
| Pellegrinelli et al. (2022) | Italy March 2019–December 2020 |
Raw (influent) | (1) Centrifugation at 4500 g for 30 min, (2) Centrifugation at 4500 g for 30 min at 4 °C, (3) Centrifugation at 1200 g for 30 min at 4 °C | (1) PEG-Dextran two-phase separation, (2) PEG precipitation chloroform purification, (3) PEG precipitation with chloroform purification | QIAgen QIAamp® MinElute Virus Spin Kit and NucliSENS EasyMAG | RT-PCR N1, N3 and ORF1ab |
(1,3) 250, (2) 80 | 2 |
| Perez-Cataluna et al. (2021) | NR | NR | (1) NR, (2) Centrifugation at 2500 g for 10 min at 4 °C | (1) Aluminum hydroxyde adsorption-precipitation, (2) PEG precipitation | NucleoSpin RNA Virus Kit | RT-qPCR N1, N2, E, IP2 AND IP4 |
200 | 1 |
| Petala et al. (2022) | Greece October 2020 – January 2021 |
Raw (influent) | pH adjustment to 4 using 2 M HCl, Centrifugation at 4000 g for 30 min | Adsorption-elution using electronegative membranes | Phenol-chloroform-based RNA extraction protocol | RT-PCR N2 and E |
200 | 5 |
| Philo et al. (2021) | USA March 2020–July 2020 |
Raw (influent after sedimentation) | NR | (1) Bag-mediated Filtration System (BMFS), (2) Skimmed milk flocculation, (3) PEG precipitation, (4) Ultrafiltration | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N1, N2 and N3 |
(1) 100, (2) 500, (3) 1000 | 1 |
| Philo et al. (2022) | October 2020–March 2021 | Raw (influent after sedimentation) | NR | Skimmed milk flocculation | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N1 and N2 |
50 | 1 |
| Pillay et al. (2021) | South Africa July 2020 – October 2020 |
Raw (influent) | Heat activation 60 °C for 90 min, Centrifugation at 3500 g for 10 min | Ultrafiltration | QIAgen QIAamp® Viral RNA mini Kit | dd-PCR N2 |
250 | 5 |
| Pino et al. (2021) | Colombia NR |
Raw (influent) | NR | (1) Flocculation with AlCl3, (2) PEG precipitation, (3) Flocculation with skimmed milk, (4) Ultrafiltration | EZNA Total RNA Kit | RT-qPCR N, E and RdRp |
200 | 2 |
| Prakash (2021) | India June 2020–July 2021 |
Raw (sewage) | (1) Centrifugation at 4700 g for 30 min, (2) Precentrigufation at 5000 rpm for 30 min, (3) NR | (1) Ultrafiltration, (2) PEG precipitation (3) PEG-dextran two-phase separation | QIAgen RNeasy PowerMicrobiome Kit | RT-qPCR N1, N2 and E |
200 – 550 | 1 |
| Qiu et al. (2022) | Canada May 2020 |
Raw (influent) | pH adjustment to 9.6–10 using 5 N NaOH, Centrifugation at 4500 g for 10 min, Removal of supernatant and pH readjustment to 7 | Centrifugal ultrafiltration | QIAgen RNeasy PowerMicrobiome Kit, MagMAX-96 Viral RNA Isolation Kit, MagMAX Viral/Pathogen Viral RNA mini Kit and ReliaPrep RNA miniprep System | RT-qPCR N1, N2, E and RdRp |
100 | 3 |
| Ramos-Mandujano et al. (2021) | Saudi Arabia June 2020 |
Raw (sewage) | NR | (1) Adsorption-elution with electronegative membranes, (2) Silica-coated magnetic nanoparticles | QIAamp® RNA mini Kit | RT-PCR N1 and N2 |
300 – 500 | 1,3 |
| Randazzo et al. (2020) | Spain March 2020 – April 2020 |
Raw (influent) and treated (secondary and tertiary effluents) | NR | Aluminum hydroxide adsorption-precipitation | NucleoSpin RNA virus Kit | RT-qPCR N1, N2 and N3 |
200 | 1 |
| Reynolds et al. (2022) | Ireland September 2020 – March 2021 |
Raw (influent) | Centrifugation at 3200 g for 5 min | Ultracentrifugation | QIAgen RNeasy PowerMicrobiome Kit | RT-qPCR and dd-PCR N1 |
200 – 225 | 2.6 |
| Robotto et al. (2022) | Italy July 2020–March 2021 |
Raw (influent) | NR | NR | Wastewater Large Volume Total Nucleic Acid Capture Kit AX9550 | RT-qPCR N1, N2 and E |
40 | 1,3 |
| Rocha et al. (2022) | USA July 2020–May 2021 |
Raw (influent) | 2.5 M MgCl2 was added at a ratio of 1:100 to a final concentration of 25 mM | Adsorption-elution with electronegative membranes | QIAgen PowerViral Kit | RT-qPCR N1 and N2 |
40 – 495 | 3,4,5 |
| Rodriguez Rasero et al. (2022) | Spain July 2020 – February 2021 |
Raw (sewage) | pH adjustment to 6 using 2 N HCl | AlCl3 precipitation | NucleoSpin RNA Virus Kit and QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N, E and IP4 |
200 | 5 |
| Roka et al. (2021) | Hungary June 2020 – October 2020 |
Raw (influent) | (1) NR, (2) Centrifugation at 4500 g for 30 min at 4 °C | (1) Skimmed milk flocculation, (2) Ultrafiltration | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N |
(1,2) 50 | 1,4 |
| Rondeau et al. (2021) | USA NR |
Raw (sewage) | Heat inactivation 60 °C for 1 h, filtering using 0.22 μm filter | Centrifugal ultrafiltration | Quick RNA miniprep Kit | RT-qPCR N1 |
40 | 3 |
| Rosiles-Gonzalez et al. (2021) | Mexico August 2020–January 2021 |
Treated (primary, biofilter and biological treatment) | (1) Filtration using a 0.45 μm cellulose-ester membrane, (2,3) Sequential filtration using 0.8, 0.65, 0.45 and 0.22 μm cellulose-ester membranes | (1) Adsorption-elution with electronegative membranes, (2) PEG precipitation, (3) Centrifugal filtration. | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N1 and N2 |
(1) 1.0 – 5.4, (2) 200 – 1000, (3) 0.6 – 1.3 | 1,3 |
| Sapula et al. (2021) | Australia NR |
Raw (influent) | (1) Centrifugation at 5000 g for 30 min at 4 °C, (2) adding MgCl2 to a final concentration of 25 mM | (1) PEG precipitation, (2) Adsorption-elution with electronegative membranes | TRIzol-phenol extraction, NucleoSpin RNA Virus Extraction Kit and RNeasy PowerWater Kit | RT-qPCR N1 and N2 |
100 | 1,3 |
| Saththasivam et al. (2021) | Qatar June 2020 – August 2020 |
Raw (influent) | Heat inactivation 56 °C for 30 min, Centrifugation 4500 g for 30 min at 4 °C | PEG precipitation | Quick RNA Viral Kits (Zymo) | RT-qPCR N1, N2 and RdRp |
200 | 2,5 |
| Scott et al. (2021) | USA August 2020 – December 2020 |
Raw (sewage) | NR | PEG precipitation | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR and dd-PCR N1 and N2 |
200 | 1 |
| Sharma et al. (2021) | India May 2020 – May 2020 |
Raw (sewage) | Chloroform was added and mixed thoroughly using a magnetic stirrer for 30 min at 4 °C, Centrifugation at 3000 g for 20 min at 4 °C | PEG-dextran phase separation | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR E and RdRp |
500 | 2 |
| Sherchan et al. (2020) | USA January 2020–April 2020 |
Raw (influent) and treated (secondary treatment, effluent) | (1) Centrifugation at 3000 g for 30 min, (2) NR | (1) Ultrafiltration, (2) Adsorption-elution with electronegative membranes | ZR Viral RNA Kit | RT-qPCR N1 and N2 |
100 – 1000 | 1 |
| Song et al. (2021) | USA April 2020 – June 2020 |
Raw (influent) | (1) Heat inactivation 60 °C for 90 min, Centrifugation at 4000 g for 30 min, filtration using 0.45 μm sterile membrane filter, (2) NR | (1) PEG precipitation, (2) Direct extraction method | QIAgen QIAamp® Viral RNA mini Kit and Zymo Quick-RNA Fecal/Soil Microbe Microprep Kit | RT-qPCR and ddPCR N1 and N2 |
50 | 1,2,6 |
| Tandukar et al. (2022) | Nepal July 2020–February 2021 |
Raw (influent, sewage) | NR | Electronegative membrane-vortex (EMV) | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N1, N2 and E |
100 | 1,3 |
| Tanhaei et al. (2021) | Iran June 2020–July 2020 |
Raw (influent) and treated (effluent) | NR | Adsorption-elution with electronegative membranes | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N and ORF1ab |
200 | 2 |
| Tanimoto et al. (2022) | Japan February 2021 – October 2021 |
Raw (influent) | Centrifugation at 10000 g for 30 min | PEG precipitation | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N |
40 | 4 |
| Thongpradit et al. (2022b) | Thailand January 2021–February 2021 |
Raw (sewage) | Centrifugation at 3000 g for 10 min at room temperature | Adsorption-elution using electronegative membranes | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N, S and ORF1ab |
100 – 400 | 2 |
| Tiwari et al. (2022) | Finland August 2020–May 2021 |
Raw (influent) | Centrifugation at 3000 g for 25 min | Ultrafiltration | Chemagic Viral300 DNA/RNA Extraction Kit | RT-qPCR E and N2 |
NR | 1 |
| Toledo et al. (2022) | USA Sept 2020 - Feb 2021 |
Raw (influent) | Centrifugation at 4600 g for 30 min at 4 °C | PEG precipitation | Promega Wastewater Large-Volume TNA Capture Kit | RT-qPCR and RT-ddPCR N1 and N2 |
45 | 1 |
| Tomasino et al. (2021) | Portugal May 2020–March 2021 |
Raw (influent) | pH adjustment to 3.5/4 using HCl, Heat inactivation 60 °C for 90 min | (1) NR, (2) Sequential centrifugations followed by PEG precipitation | QIAgen RNeasy Powersoil Total RNA, QIAgen RNeasy PowerMicrobiome Kit and IDEXX DNA/RNA Magnetic Bead Kit | RT-qPCR N1 and N2 |
(1) 10 – 80, (2) 35 | 3 |
| Torii et al. (2021) | Japan NR |
Raw (influent) | (1) Centrifugation at 3500 g for 15 min, (2) filtration using a through 0.45 μm cellulose-ester membrane, (3) Centrifugation at 3500 g for 5 min | (1) Ultracentrifugation, (2) EMV, (3) PEG precipitation | QIAgen QIAamp® Viral RNA mini Kit and Acid guanidium thiocyanate-phenol-chloroform extraction using TRIzol protocol | RT-qPCR N1, N2 and N3 |
40 – 50 | 2 |
| Torii et al. (2022) | Japan July 2020–October 2020 |
Raw (influent) | (1,2) Centrifugation at 3500 g for 5 min, (3) Centrifugation at 4700 g for 30 min at 4 °C, (4) filtration using a 0.2 μm hydrophilic polytetrafluoroethylene membrane (Millipore), (5) NR | PEG precipitation | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N1 and N2 |
41 | 1,2 |
| Trottier et al. (2020) | France May 2020–July 2020 |
Treated (effluent) | Centrifugation at 4500 g for 30 min at 4 °C, | Centrifugal filtration | NucleoSpin RNA Virus Kit | RT-qPCR N1, N2 and RLP27 |
50 | 3 |
| Trujillo et al. (2021) | NR | NR | Heat inactivation 60 °C for 60 min, filtration using a 0.22 μm filter | PEG precipitation | TRIzol-chloroform protocol | RT-qPCR N1 |
40 | 3 |
| Vallejo et al. (2022) | Spain March 2020 – May 2020 |
Raw (influent) | Centrifugation at 4000 g for 30 min, filtration using 0.22 μm membranes | Ultrafiltration | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N |
100 | 4.5 |
| Wehrendt et al. (2021) | Argentina April 2021–May 2021 |
NR | (1) Centrifugation at 12000 g for 1 h at 4 °C, (2) pH adjustment to 6–7 | (1) PEG precipitation, (2) Centrifugation with PAC | High Pure Viral Nucleic Acid Kit and Viral Nucleic Extraction Kit II | RT-qPCR N and ORF1 |
(1) 200, (2) 40 | 1,3,4 |
| Westhaus et al. (2021) | Germany April 2020 |
Raw (influent after sand trap) and treated (activated sludge) | Centrifugation at 4700 g for 30 min | Ultracentrifugation | NucleoSpin RNA virus Kit | RT-qPCR N, E and RdRp |
45 | 2 |
| Whitney et al. (2021) | USA NR |
Raw (influent) | NR | NR | 4S-column and 4S-Milk-of-Silica | RT-qPCR N1 |
40 | 1 |
| Wu et al. (2022) | USA January 2020 – Ma 2020 |
Raw (influent) | Heat inactivation 60 °C for 90 min, filtration using a 0.2 μm sterile membrane filter | PEG precipitation | TRIzol-chloroform protocol | RT-qPCR N1 and N2 |
40 | 4,5 |
| Xiao et al. (2022) | USA March 2020 – June 2020 |
Raw (influent) | Heat inactivation 60 °C for 1 h, filtration using a 0.2 μm vacuum-driven filter |
Centrifugal ultrafiltration | NR | RT-qPCR N1 and N2 |
15 | 5 |
| Xu et al. (2021) | Hong Kong June 2020 – September 2020 |
Raw (influent and sewage) | Heat inactivation 60 °C for 30 min, Centrifugation at 4750 g for 30 min | Ultrafiltration | TRIzol Plus RNA Purification Kit | RT-qPCR N |
50 – 90 | 1 |
| Yanac et al. (2022) | Raw (influent) and treated (primary sludge, secondary effluent, final effluent) | (1) Cheesecloth and low-protein binding 0.45 and 0.2 μm 47-mm Supor-200 membrane disc filters, (2) NR |
(1) Ultrafiltration, (2) Skimmed milk flocculation | QIAgen RNeasy PowerMicrobiome Kit and MagMAX Microbiome Kit | RT-qPCR N1 and N2 |
120 | 1,4,5 | |
| Yaniv et al. (2021) | Israel November 2020–March 2021 |
Raw (influent) | Shaken and mixed for 2 min manually and left standing 15 min to large particle settlement | Ultrafiltration | NucleoSpin RNA Extraction Kit | RT-qPCR N1, N2, N3 and N4 |
2000 – 5000 | 1,5 |
| Zhang et al. (2022) | Australia August 2020 – September 2020 |
Raw (influent) | NR | Adsorption-elution with electronegative membranes | QIAgen RNeasy PowerWater Kit | RT-qPCR N and E |
100 | 2 |
| Zhao et al. (2022) | USA September 2020 – August 2021 |
Raw (influent) | NR | PEG precipitation | QIAgen QIAamp® Viral RNA mini Kit | RT-ddPCR N1 and N2 |
NR | 4 |
| Zheng et al. (2022) | Hong Kong September 2020 – November 2020 |
Raw (influent and sewage) | Heat inactivation 60 °C for 30 min | (1) Ultracentrifugation, (2) PEG precipitation, (3) AlCl3 flocculation, (4) MgCl2 flocculation, (5) Ultracentrifugation 10 kDa, (6) Ultracentrifugation 30 kDa, (7) Membrane adsorption with AlCl3, (8) Adsorption-elution using electronegative membranes, (9) Combination of centrifugation and ultracentrifugation, (10) AlCl3 precipitation, (11) Membrane adsorption | QIAgen QIAamp® Viral RNA mini Kit and TRIzol Plus RNA Purification Kit | RT-qPCR N1 |
30 – 1000 | 1 |
| Zhu et al. (2022) | Japan August 2020 – February 2021 |
Raw (influent) | NR | Ultracentrifugation | QIAgen QIAamp® Viral RNA mini Kit | RT-qPCR N1 |
40 | 4,5 |
Note: NR stands for Not Reported.