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Molecular detection of SARS-CoV-2 from wastewater

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Requisition Forms

Reference Details


RT qPCR surveillance and quantitation of SARS-CoV-2 from wastewater

Test Category:
Molecular Detection
Illnesses and Diseases:
  • Respiratory tract infections
  • COVID-19

Raw sewage, primary influent, primary sludge, effluent, passive sampler media (eluate, filter, membrane)

Collection Method:

Irrespective of sample matrix, 500 mL of composite or grab samples should be sent to PHAC NML as per the shipping guidance below. Ideally, the sample should be maintained at 4°C during collection. The impact of common water/wastewater preservatives has not been assessed and submitters are discouraged from sending preserved samples.  Eluate or whole membranes from passive sampling devices may be submitted provided they are packaged according to the shipping guidance below.

Specimen Processing, Storage and Shipping:

Samples should be sealed in a leak proof primary vessel (no glass) within a sealed secondary container. Secondary container packaging should contain an appropriate amount of absorbent material capable of absorbing the total volume of liquid being shipped in case of leak or spill. All samples should NOT BE FROZEN at any time prior to, or during shipment and should be maintained at 4°C with frozen cold packs preferred, if unavailable wet ice may be used, but must be double bagged. All samples should be specifically marked with sample collection site and date collected and identified as environmental or wastewater samples. Please ship samples and cold packs contained within temperature preserving shipping pack (styrofoam or hard shell cooler), contained within appropriately sized and labelled cardboard overpack.

Transportation of Dangerous Goods:

Shipping of specimens shall be done by a TDG certified individual in accordance with TDG regulations. For additional information regarding classification of specimens for the purposes of shipping, consult either Part 2 Appendix 3 of the TDG Regulations or section 3.6.2 of the IATA Dangerous Goods Regulations as applicable.

Patient Criteria:


Accompanying Documentation:

Requisition for Detection of Respiratory Viruses from Wastewater


All samples must be addressed to the Canadian Science Centre for Human and Animal Health at 1015 Arlington St. in Winnipeg, MB for mandatory sample receiving protocols.

Sample Rejection Criteria:  For preservation of sample integrity and data quality, our samples must abide by stringent collection timelines. As such, the acceptable time between sample collection and laboratory receiving is a maximum of 3 weeks. Any samples not sent within the aforementioned shipping standards, i.e. leaking or improperly packaged, may be subject to rejection by NML or respective carriers. Sample will be immediately rejected for testing if greater than 20% of a sample (100mL) has leaked into the secondary container/bag, if samples do not arrive with individual labelling or identifying markers, or if samples arrive unmarked without listing on an accompanying sample requisition form.

Methods and Interpretation of Results:

Quantitative analysis within limits of quantification of CDC SARS-CoV-2 RT qPCR targets N2 and N200 universal SARS-CoV-2 target (Fuzzen 2023). All results are analysed for Pepper Mild Mottle Virus (PMMoV) endogenous control detection for the purpose of signal normalization and QA.

Turnaround Time:

3-5 calendar days.

Phone #: 204-789-6506
Fax: 204-318-2221

Nucleic Acid Amplification Tests (NAATs)

Fuzzen M., et al. An improved method for determining Frequency of multiple variants of SARS-CoV-2 in wastewater using qPCR assays. Sci Total Environ. 2023; 881: 163292 doi: 10.1016/j.scitotenv.2023.163292

The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Bustin SA, et al. 2009 Apr;55(4):611-22. doi: 10.1373/clinchem.2008.112797.