Table 3.
Detection and quantification of viruses in wastewater and sludge samples.a
| Wastewater sample | Viruses | Concentration/pre-treatment method | Nucleic acid extraction | Virus detection/quantification | Reference |
|---|---|---|---|---|---|
| Wastewater treatment samples: (a) Effluent of natural oxidizing pond (b) Effluent of rotating biodisk |
AiV Genotype B | Beef extract and AlCl3 method followed by precipitation with PEG | Automatic extractor NucliSENS® EasyMag™ | RT-PCR | Ibrahim et al., 2017 |
| Raw sewage | Oncogenic viruses: HPV HPyV HHV MMTV |
Elution with glycine | Automatic extractor NucliSENS® EasyMag™ | Combined multiplex PCR and bead-based Luminex technology | Di Bonito et al., 2017 |
| Influent wastewater | HAdV EV RoV NoV GI NoV GII |
VIRADEL | QIAamp DNA Blood Mini Kit | Combined real time PCR (qPCR) and multiplex Luminex xMAP assay | Hamza et al., 2014 |
| Combined wastewater; Graywater | HAdV NoV GI NoV GII |
Ultrafiltration followed by elution with sodium polyphosphate solution | QIAamp DNA Blood Maxi Extraction Kit | (a) For HAdV: qPCR and TaqMan assay; Digital droplet PCR (ddPCR) (b) For Nov GI and NoV GII: RT-qPCR; RT-ddPCR |
Jahne et al., 2020 |
| Influent wastewater | PV1 | Primary: Filtration with electropositive filter and elution with beef extract and glycine Secondary: (a) beef extract-Celite (b) ViroCap flat disc filter (c) concentrating Pipette (d) PEG/NaCl precipitation (e) skimmed-milk flocculation |
– | – | Falman et al., 2019 |
| (a) Influent municipal wastewater; (b) Effluent municipal wastewater |
HEV HAdV 40 Nov GII Porcine Adenovirus |
(a) VIRADEL (b) Precipitation with PEG |
QIAamp Viral RNA Mini Kit | RT-PCR | Masclaux et al., 2013 |
| Effluent | EV RoV |
Adsorption-elution method (Mg-method and Al-method) | ZR Viral RNA kit | RT-qPCR | Osuolale and Okoh, 2017 |
| Hospital wastewater treatment plant samples: (a) Influent (b) Effluent of sedimentation tank (c) Final effluent (after chlorination) |
RoV-A HAdV NoV GI NoV GII HAV |
Adsorption-elution using electronegative membrane | QIAamp Viral RNA Kit | (a) Conventional PCR, RT-PCR (b) qPCR |
Prado et al., 2011 |
| Municipal wastewater treatment samples: (a) Pre-UV treatment (b) Post-UV treatment |
NoV RoV Sapovirus Astrovirus JC Virus HAdV EV Reovirus |
Adsorption-elution using NanoCeram disc filters | MagaZorb total RNA Prep kit | qPCR | Qiu et al., 2018 |
| Municipal wastewater treatment samples: (a) Influent (b) Effluent of Secondary Treatment (c) Reclaimed water |
HAdV JCPyV RoV-A |
(a) For Influent and Secondary Effluent: Concentration with Celite (b) Reclaimed water: Ultrafiltration |
QIAmp Viral RNA Mini Kit | RT-qPCR | Prado et al., 2019 |
| Wastewater treatment samples: (a) Influent (b) Effluent |
HAdV HPyV HTtV |
Ultrafiltration | Qiagen DNeasy Blood and Tissue kit | qPCR | Sidhu et al., 2018 |
| Wastewater treatment samples: (a) Influent (b) Effluent of water treatment ponds (c) Effluent of UASB reactor (d) Sludge from water treatment pond (e) Sludge from UASB reactor |
NoV RoV PMMV EV |
Sludge samples: ASTM 4994-19 Wastewater samples: (a) For EV: Dilution with beef extract solution, centrifugation and filtration through protein-treated filter (b) For q-PCR: Adsorption- elution |
AllPrep DNA/RNA Mini Kit | (a) For NoV, RoV, PMMV: RT-qPCR (b) For EV: Cell Culture |
Symonds et al., 2014 |
| Municipal wastewater treatment samples: (a) Influent (b) Sludge |
Virus type not determined (quantification only) | Elution with solutions (a) Beef extract (b) Sodium pyrophosphate (c) Glycine (d) Potassium citrate (e) Lysozyme |
– | (a) Epifluorescence Microscopy (EFM) using SYBR Green I as stain (b) Transmission Electronic Microscopy (TEM) (a) Pulsed-field Gel Electrophoresis |
Wu and Liu, 2009 |
| Influent wastewater | HAdV JCPyV |
Concentration with Phosphate-buffered saline (PBS) solution | For qPCR: QIAamp Viral RNA mini Kit | (a) Immunofluorescence assay (b) Plaque assay (c) Tissue culture infectious dose-50 (d) qPCR |
Calgua et al., 2011 |
| Influent wastewater Primary settled wastewater Secondary settled wastewater Activated sludge Effluent |
Virus type not determined (quantification only) | Ultrasonication | – | Flow cytometry (FCM) | Ma et al., 2013 |
| Activated sludge | Virus type not determined (quantification only) | Addition of dispersants Tween 80, sodium pyrophosphate, and sodium cholate; Ultrasonication |
– | Flow cytometry | Brown et al., 2015 |
| Activated sludge mixed liquor | Virus type not determined (quantification and determination of viral size distribution only) | Sonication | PFGE Method | PFGE | Otawa et al., 2007 |
| Sludge samples (influent and effluent of anaerobic digesters) | Enteroviruses Coronavirus HKU1, Klassevirus, Cosavirus Parechovirus |
Precipitation with PEG | Qiagen Viral RNA extraction kit | PCR; High-throughput Sequencing |
Bibby and Peccia, 2013 |
| Sludge sewage samples | Enteric viruses | EVE | Sepa-gene RV-R | RT-PCR | Sano et al., 2003 |
| Sewage sludge samples | HAdV RoV-A NoV GII HAV |
Ultracentrifugation; Beef Extract Elution |
QIAamp Viral RNA Kit | RT-qPCR qPCR |
Prado et al., 2014 |
| Primary sludge samples Activated sludge samples Thickened sludge samples |
EV | Beef Extract elution with sonication; Precipitation with PEG |
RNeasy plant mini kit | RT-PCR | Monpoeho et al., 2000 |
| Raw sludge samples Treated sludge samples |
Mammalian orthoreovirus | Adsorption elution; Organic Flocculation |
QIAamp Viral RNA Kit | ICC-RT-qPCR; Plaque Assay |
Gallagher and Margolin, 2007 |
Note: aData for SARS-CoV-2 are reported in Table 4.