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Wastewater Surveillance via Nucleic Acid |
Wastewater surveillance is a useful tool for monitoring the regional spread of infectious diseases before an outbreak occurs.
How does wastewater surveillance work?
Wastewater disease surveillance data can be used to monitor the regional or city-level spread of pathogens. It can also be used to analyze targeted sites to identify asymptomatic carriers, capture emerging mutations, and respond to possible infections. With the emergence of SARS-CoV-2 infections, multiple organizations worldwide worked on SARS-CoV-2 surveillance methods using community sampling and tracking of viral nucleic acids in wastewater.
Methods of detection and surveillance for pathogens including viruses, gram-positive, and gram-negative bacteria are outlined on this page. The same nucleic isolation technology can also be used with samples such as wastewater, sludge, fresh and salt water.
MagMAX kits for wastewater surveillance
Automated, high-throughput nucleic acid extraction from wastewater samples for use in applications such as real-time PCR and next-generation sequencing.
Automated, high-throughput virus enrichment and nucleic acid extraction from wastewater samples for use in applications such as real-time PCR and next-generation sequencing.
Explore: MagMAX Wastewater Ultra Nucleic Acid Isolation Kit, with Virus Enrichment
Wastewater surveillance testing workflow
Wastewater concentration
Viral concentration allows for improved detection and downstream analysis. Viral enrichment with Dynabeads magnetic beads is designed for efficient and advanced concentration.
Automated sample processing
KingFisher instruments paired with MagMAX wastewater kits allow for a completely automated nucleic acid isolation format without inconvenient centrifugation, filtration, or bead beating steps.
Nucleic acid purification
From the eluate containing concentrated viral particles, viral nucleic acid can be purified using the Applied Biosystems MagMAX Wastewater Ultra Nucleic Acid Isolation Kit.
Wastewater surveillance sample concentration protocols
| Wastewater starting volume | Sample details | Kit reagents sufficient for | KingFisher scripts available | Protocol time | Purchase this kit |
|---|---|---|---|---|---|
| 10 mL | Concentration of 10 mL wastewater using Dynabeads followed by nucleic acid isolation | 100 preps | Flex, Duo Prime, APEX | 90 minutes | MagMAX Wastewater Ultra Nucleic Acid Isolation Kit, with Virus Enrichment |
| 200 μL | 200 μL wastewater pre-concentrated using a preferred method, e.g., ultracentrifugation | 100 preps | Flex, Duo Prime, APEX | 45 minutes | MagMAX Wastewater Ultra Nucleic Acid Isolation Kit |
| 1 mL* | A. 1 mL non-concentrated wastewater B. Wastewater pre-concentrated to 1 mL volume using a preferred method, e.g., precipitation | 20 preps | Flex, Duo Prime, APEX | 45 minutes | |
| 50–500 mL | Concentration of 50–500 mL wastewater using filtration followed by nucleic acid isolation | 20 preps | Flex, Duo Prime, APEX | 90 minutes | |
| Sludge | 200 mg sludge | 20 preps | Flex, Duo Prime, APEX | 45 minutes |
*works for 1–2 mL, but 1 mL is the most sensitive.
Automate MagMAX Kits using KingFisher instruments
Benefits of KingFisher instruments include
- Various throughputs—Process 6–96 samples per run depending on the instrument model
- Interchangeable formats—Process a wide range of input volumes
- Protocol customization—Easily edit, modify, or create new protocols
- Optimized reagents—Compatible with multiple magnetic-bead reagents
Explore: KingFisher instruments
Download: Validated software automation scripts for KingFisher purification instruments
Wastewater surveillance resources
Accurate quantification of SARS-CoV-2 in wastewater samples
A complete workflow for wastewater surveillance from sample collection to SARS-CoV-2 quantification is shown in this study. Benefits of this workflow include fast time-to-results, great flexibility, and multiplexing capacity. Microfluidic array plate (MAP) technology enables effortless and consistent sample digitization, while multiplexing allows for detection of up to 4 targets in a single reaction.
Study: Accurate quantification of SARS-CoV-2 in wastewater samples
Protocols for wastewater sample concentration
Optimized protocol for the enrichment of SARS-CoV-2 in wastewater samples. Large sample volumes of wastewater are dilute. Thus, they must be enriched for intact viral presence for pathogen detection via qPCR. Applied Biosystems MagMAX Wastewater Ultra Nucleic Acid Isolation Kits offer a whole host of advantages over traditional wastewater surveillance methods. Isolate nucleic acids from concentrated wastewater in as little as 45 minutes. Our viral enrichment protocol using magnetic beads enables concentration and nucleic acid isolation in less than 2 hours.
Multiple workflow options for detection of SARS-CoV-2 in wastewater
Five robust workflows were developed for rapid concentration and qPCR analysis of SARS-CoV-2 from different volumes of wastewater: low (1 mL), medium (10 mL), and large (50 mL). The protocols described here are extensions of those developed in previous studies and have been optimized for processing multiple sample types. The protocols include a fully automated workflow to enable high-throughput processing; this workflow has been successfully used in the field.
Study: Multiple workflow options for detection of SARS-CoV-2 in wastewater
Nucleic acid purification resources
Protocols for wastewater nucleic acid purification
This automated protocol has been optimized for the concentration of SARS-CoV-2 and other viral targets in wastewater samples since these samples are highly diluted and pre-enrichment is needed to help improve the detection of pathogens. The method we describe here can be used for downstream nucleic acid extraction after enrichment for viral/pathogen targets with Applied Biosystems MagMAX Wastewater kits and requires just 90 minutes of processing time. Using Applied Biosystems Dynabeads Wastewater Virus Enrichment beads for SARS-CoV-2 and other viral targets from wastewater samples followed by bead-based nucleic acid extraction is designed to be simple, fast, and reliable.
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For Research Use Only. Not for use in diagnostic procedures.
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