Silica based kit solutions for wastewater surveillance
Please find below our silica based kit suggestions for wastewater surveillance.
NucleoSpin RNA Stool
- A single spin kit for small scale purposes
- Advanced inhibitor removal technology
- Compatible with all common downstream applications, including NGS
Recommendations for sample concentration from wastewater
Concentration of viral particles are essential for efficient nucleic acid isolation from water samples. However, due to their size virus particles cannot be efficiently concentrated on a membrane without a specific treatment. Therefore, we summarize the three most common sample pretreatment recommendations for viral particle concentration from water samples prior nucleic acid isolation.
Filtration and concentration of viral particles using electronegative membranes
Methods for concentrating viral particles on filter membranes have been described in Ahmed et al. 2020 and other publications. For filter membranes we recommend a bead-based mechanical lysis for optimal results. Click here to read more about wastewater concentration with electronegative membranes.
Ultrafiltration
This approach for viral concentration has been described by Ahmed et al. 2020 and Medema et al. 2020 utilizing ultrafiltration and 100–200 mL wastewater samples. Larger particles such as debris or bacteria are removed by centrifugation (e.g. 4600 - 4700 x g, 30 min). It is also possible to use standard filtration methods to remove debris. The supernatant (centrifugation) or cleared sample (standard filtration) is subjected to ultrafiltration (e.g. Centricon Plus-70 centrifugal ultrafilter units 10 kDa, Merck; 1500 - 3500 x g for 15 min). Subsequently, nucleic acids can be isolated from the recovered, liquid concentrate using either NucleoSpin RNA Stool or NucleoMag DNA/ RNA Water following the standard protocols, however the mechanical lysis step can be omitted
Precipitation
Different methods can be used for precipitating viral particles. Randazzo et al. 2020 describe an aluminum hydroxide adsorption-precipitation basedmethod, Wu et al. 2020 a PEG-precipitation based approach. Precipitation methods result in a pellet that can be used for further extraction of nucleic acids. To ensure efficient removal of PCR inhibitors we recommend NucleoSpin RNA Stool or NucleoMag DNA/RNA Water for nucleic acid extraction, although NucleoSpin RNA Virus Kit has been successfully used in combination with precipitation by Randazzo et al. 2020.
NucleoSpin RNA Virus
quick and efficent lyis for RNA viruses
designed for the rapid preparation of highly pure nucleic acids
no cross contamination due to closed systems
Success story - DyeNA Genetics - Wastewater surveillance with the NucleoSpin RNA Virus kit
Read more about the key features of the DyeNA Genetics workflow together with the NucleoSpin RNA Virus kit.
"The NucleoSpin RNA Virus kit allows a very sensitive, fast and straight forward purification of SARS-CoV-2 from wastewater."
- concentration and purifiaction of viral RNA within less than 2 hours
- results are reported usually on the same day of sample arrival
- very sensitive, reliable and reproducible data
- the method has a predictive early warning function from 7 to 14 days
Application data
Quantification of SARS-CoV-2 from wastewater from different treatment plants
Determination of the quantity of SARS CoV-2 with our qPCR test system in the wastewater (24-hour composite sample from the influent) of various wastewater treatment plants with different numbers of ill persons in the connected municipality. Shown are the number of infected persons (grey) and the total amount of virus genes (green) that entered the plant within 24 hours through wastewater inflow.
qPCR coronavirus monitoring of a municipal sewage treatment plant in northern Bavaria.
The incidence values of the municipality (grey) and the CoV-2 total virus quantity (green), which flows into the sewage treatment plant within 24 hours, are shown. The qPCR readings show changes in the incidence of infection 7 - 14 days earlier and thus fulfil the function of an early warning or all-clear system. This can be clearly seen around 21.04.2021. From this time onwards, the viral load in the wastewater is already reduced, and approximately 10 - 14 days later (02.05. - 07.05.2021) the downward trend also begins in the incidence values. The trend reversal or the beginning of the fourth well was also precisely determined, as was the renewed, strong increase in the infection figures on 22.08.21, 18.09.21, 19.10.21 and 26.12.21, which were indicated early on.
Recommendations for sample concentration from wastewater
Sample concentration on the INNOVAPREP Concentrating Pipette Select
The INNOVAPREP Concentrating Pipette Select offers in combination with the NucleoSpin RNA Virus kit an efficient and inhibition-free nucleic acid purification from synthetic wastewater samples. Click here to read more about this sample concentration method.
Filtration and concentration of viral particles using electronegative membranes
Methods for concentrating viral particles on filter membranes have been described in Ahmed et al. 2020 and other publications. For filter membranes we recommend a bead-based mechanical lysis for optimal results. Click here to read more about wastewater concentration with electronegative membranes.
Ultrafiltration
This approach for viral concentration has been described by Ahmed et al. 2020 and Medema et al. 2020 utilizing ultrafiltration and 100–200 mL wastewater samples. Larger particles such as debris or bacteria are removed by centrifugation (e.g. 4600 - 4700 x g, 30 min). It is also possible to use standard filtration methods to remove debris. The supernatant (centrifugation) or cleared sample (standard filtration) is subjected to ultrafiltration (e.g. Centricon Plus-70 centrifugal ultrafilter units 10 kDa, Merck; 1500 - 3500 x g for 15 min). Subsequently, nucleic acids can be isolated from the recovered, liquid concentrate using either NucleoSpin RNA Stool or NucleoMag DNA/ RNA Water or NucleoSpin RNA Virus following the standard protocols, however the mechanical lysis step can be omitted
Precipitation
Different methods can be used for precipitating viral particles. Randazzo et al. 2020 describe an aluminum hydroxide adsorption-precipitation basedmethod, Wu et al. 2020 a PEG-precipitation based approach. Precipitation methods result in a pellet that can be used for further extraction of nucleic acids. To ensure efficient removal of PCR inhibitors we recommend NucleoSpin RNA Stool or NucleoMag DNA/RNA Water for nucleic acid extraction, although NucleoSpin RNA Virus Kit has been successfully used in combination with precipitation by Randazzo et al. 2020.