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Virus Detection in Wastewater Through PCR Tests

Virus Detection in Wastewater Through PCR Tests
Analytik Jena’s solution to monitor viruses and bacteria in wastewater samples based on PCR tests. (Credit: Analytik Jena)

Analytik Jena’s innovative solution can monitor biological parameters such as viruses and bacteria in wastewater samples based on PCR tests. The accurate test results can be obtained within only three to four hours.

Covid-19 showed the entire world how vital epidemiological surveillance and getting an early handle on the spread of infections are. These tasks fall to both governments and multilateral institutions and have several and complex fronts. One of them in particular has been catching the attention of experts due to its low costs and great benefits: wastewater analysis based on PCR tests.

Genetic information, including virus particles and other germs, is detectable in wastewater above a certain level of concentration. This resource has been gaining ground with Covid-19 monitoring in sewage treatment plants playing a major role in this development. 

As a matter of fact, on March 17th the European Union issued a recommendation which stipulates that the Member States should regularly monitor wastewater for SARS-CoV-2 viral loads in cities with over 150,000 inhabitants. Not least, putting wastewater to use as a data source can lead to not only public health gains but also ecological and infrastructure ones.

Robert Möller, Analytik Jena program manager, explained:

“This information can be used to build on the critical infrastructure, especially with a view to water recycling, in order to protect the population and avert threats.”

Full Process Chain of Wastewater Monitoring

In cooperation with its parent company Endress+Hauser, Analytik Jena is the only European company that provides the full process chain of wastewater monitoring. It goes from sampling in the treatment plant to filtration and scalable nucleic acid extraction. 

Both Endress+Hauser and Analytik Jena worked with the Ruhr district located Emschergenossenschaft/Lippeverband (EGLV), Germany’s largest water management association, on a common approach to virus detection. A fully functional and tested workflow was made available by spring 2021.

Here are the following steps:

Step 1: Sampling in the Treatment Plant

The fully automatic Endress+Hauser Liquistation CSF48 takes the samples, with the mobile CSP44 variant in use upstream.

Step 2: Concentration of Target Organisms

The sample is then concentrated in order to make the detection of the target organisms possible. Subsequent to electronegative filtration, the virus fragments and particles are removed from the filter membrane using the SpeedMill PLUS. This process is several times faster than the usual time-consuming ultrafiltration, which can only process a handful of samples at a time.

Step 3: Sample Isolation in the Laboratory

For this step, Analytik Jena pipetting robots are used for the isolation: either the InnuPure C16 touch, which processes up to 16 samples, or the CyBio FeliX® for 96 samples. Both offer ready-to-use protocols which in combination with the relevant extraction kits automatically purify DNA or RNA from the samples.

Step 4: Detection

The qTOWER3 system takes on detection using fluorescence measurement. Within the polymerase chain reaction (PCR), the targeted DNA or cDNA segment is copied and marked with fluorescence. The more often the series of letters in question appears, the stronger the signal becomes in the real-time PCR process and the clearer it can be determined how high the viral load actually is.

Even non-specialists can use Analytik Jena’s solution. The accurate test results can be obtained within just three to four hours. Dr. Christina Meinert-Berning from the water-management company Ruhrverband said:

“The workflow is easy to manage, especially concerning filtration, and this is crucial from the perspective of the routine laboratory.”

The workflow is currently in use at several universities and in commercial laboratories. Round robin tests—interlaboratory tests which can be measurements, analyses or experiments and are performed independently several times—have shown that the Analytik Jena solution delivers specific and sensitive results.

The Past and the Future

Using samples from wastewater to analyze biological parameters through genetic information extraction was something unthinkable just a few years ago, and not only because of the technology and efficiency involved in the laboratory work. 

Taking samples from sewage treatment plants is deemed to be difficult. Silvio Beier, Professor of Technologies for Urban Material Flow Use at the Bauhaus University of Weimar, said:

“The influence of factors such as dilution effects and features peculiar to the catchment area of sewage treatment plants must be taken into account in order to reliably map the infection process.”

Other Applications for Legionella

Beier is the head of the Corona Monitoring in Thuringia (CoMoTH) project, which involves 23 sewage treatment plants covering around 40% of the state of Thuringia’s population—and has Analytik Jena as a participating partner. The goal is to explore the possibilities for comprehensive and permanent wastewater monitoring for SARS-CoV-2 in the state. Dr. Robert Möller from Analytik Jena explained:

“Our solution can be used to detect the spread of other pathogens among the population.” 

These early warning and monitoring systems could be applied to various viruses (such as hepatitis, polio, noroviruses, and influenza viruses), bacterial pathogens (like salmonella, clostridia, legionella and cholera) and also the spread of antibiotic-resistant germs—a growing concern within medical and scientific communities.

That is why, in addition to SARS-CoV-2 monitoring guidance, Analytik Jena already has an applications manual for legionella and one for other relevant markers in wastewater, such as norovirus and fecal markers.

(Photo credit: Analytik Jena).