Water SARS-CoV-2 RT-PCR Test

Raw wastewater that has not been chemically treated.

The Water SARS-CoV-2 RT-PCR Test is a reverse transcription quantitative polymerase chain reaction (RT-qPCR) test that detects and quantifies the viral RNA in an appropriately prepared wastewater sample.

When used with the 105 mL concentration protocol, the lowest concentration of SARS-CoV-2 in wastewater that can be detected by the Water SARS-CoV-2 RT-PCR Test is approximately 1 gene copy/1 mL.

When used with the end-to-end IDEXX procedure, the IDEXX test has been validated to provide highly consistent recovery of SARS-CoV-2 in wastewater.

The IDEXX-validated protocol concentrates the SARS-CoV-2 virus from 105 mL of wastewater. Laboratory staff may analyze larger or smaller water volumes by performing the basic procedure scaled for different input volumes with appropriate centrifugation equipment.

The Water SARS-CoV-2 RT-PCR Test analyzes 5 µL of extracted RNA prepared from a wastewater concentrate sample, although we recommend having additional volume on hand to account for pipetting loss and duplicate samples.

IDEXX provides an example concentration protocol based on centrifugation and polyethylene glycol (PEG) precipitation that has been validated to provide consistent results. This protocol can be modified to meet your specific laboratory needs. Other concentration methods may also be used, such as ultrafiltration, manifold filtration, and others, and customers have successfully used these concentration methods with IDEXX reagents. These should be validated to verify performance.

Maintaining a cold sample temperature helps to prevent sample degradation and ensure efficient precipitation of the SARS-CoV-2 virus.

The Water SARS-CoV-2 RT-PCR Test has been validated for use with the Water DNA/RNA Magnetic Bead Kit for extraction of nucleic acids, including SARS-CoV-2 RNA, from a wastewater concentrate. Other extraction kits can also be used with the test, and customers have successfully used other kits with the IDEXX RT-qPCR reagents. 

Wastewater is an extremely complex matrix, and wastewater surveillance requires extensive control materials. Please refer to the IDEXX Materials and Procedures Validated for Quantification of SARS-CoV-2 in Wastewater report for a full description of the end-to-end procedure and the required and recommended controls.

Please refer to the product inserts for recommended storage conditions at time of product receipt and after reconstitution/first use.

The U.S. Centers for Disease Control and Prevention (CDC) indicates that a biosafety level 2 (BSL-2) laboratory with additional precautions can be used for environmental sample testing, including handling and processing wastewater samples associated with SARS-CoV-2. Specific guidance is available from the U.S. CDC: Interim Laboratory Biosafety Guidelines for Handling and Processing Specimens Associated with Coronavirus Disease 2019 (COVID-19).⁴

Consult your local health authority for specific recommendations.

Follow local health authorities’ recommended procedures for handling and processing of wastewater samples associated with SARS-CoV-2. One source of information is published by the U.S. CDC: Interim Laboratory Biosafety Guidelines for Handling and Processing Specimens Associated with Coronavirus Disease 2019 (COVID-19).⁴

Follow all local regulatory and safety guidelines. 

An optional heat treatment process may be used to inactivate samples through pasteurization, if desired. See the product insert for an example. 

A pasteurization step may reduce SARS-CoV-2 RNA detection. Please contact an IDEXX account representative for additional information.

Dispose of all samples and materials in accordance with local regulatory guidelines.

The Water SARS-CoV-2 RT-PCR Test is not U.S. EPA approved for testing for SARS-CoV-2 in wastewater. The U.S. EPA does not currently regulate SARS-CoV-2 wastewater testing or approve testing methods for this purpose. Testing for SARS-CoV-2 in wastewater may or may not be regulated by your local and/or state authorities. Please check for regulations that may apply in your area.

Yes, the Water SARS-CoV-2 RT-PCR Test is compatible with, or includes, all of the controls and processes recommended by the CDC for submission of data to the NWSS. Please refer to the IDEXX Materials and Procedures Validated for Quantification of SARS-CoV-2 in Wastewater report for a full description of the end-to-end procedure, which includes all of the recommended and required controls.