Detection of viruses in wastewater using PCR tests

Analytik JenaThe innovative solution can monitor biological parameters such as viruses and bacteria in wastewater samples based on PCR testing. Accurate test results can be obtained in just three to four hours.

The Covid-19 The pandemic has shown the world how vital epidemiological surveillance and early control of the spread of infections are. These tasks fall to both governments and multilateral institutions and involve several complex fronts. One in particular has caught the attention of experts due to its low costs and great advantages: wastewater analysis based on PCR tests.

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

In fact, on March 17, the European Union issued a recommendation which states that member states must regularly monitor wastewater for SARS-CoV-2 viral loads in cities with more than 150,000 inhabitants. Last but not least, using wastewater as a data source can lead not only to public health gains, but also to ecological and infrastructure gains.

Robert Möller, Program Manager Analytik Jena, explained:

“This information can be used to strengthen critical infrastructure, including for water recycling, to protect people and prevent threats.”

Complete wastewater monitoring process chain

In collaboration with its parent company Endress+Hauser, Analytik Jena is the only European company that provides the complete process chain for wastewater monitoring. This ranges from sampling in wastewater treatment plants to filtration and the evolutionary extraction of nucleic acids.

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 in spring 2021.

Here are the following steps:

Step 1: Sampling in the treatment plant

All automatic Endress+Hauser Clearance CSF48 takes the samples, with the CSP44 mobile variant used upstream.

Step 2: Concentration of target organisms

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

Step 3: Isolation of samples in the laboratory

For this step, Analytik Jena pipetting robots are used for 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 that, in combination with the appropriate extraction kits, automatically purify DNA or RNA from samples.

Step 4: Detection

The qTOWER3 system supports detection using fluorescence measurement. In the polymerase chain reaction (PCR), the targeted DNA or cDNA segment is copied and fluorescently labeled. The more often the series of letters in question appears, the stronger the signal becomes in the real-time PCR process and the more clearly it is possible to determine the real level of the viral load.

Even non-specialists can use Analytik Jena’s solution. Accurate test results can be obtained in just three to four hours. Dr. Christina Meinert-Berning of the Water Management Society Ruhrverband said:

“The workflow is easy to manage, especially when it comes to filtration, which is crucial from a routine lab perspective.”

The workflow is currently in use at several universities and in commercial labs. Interlaboratory tests, interlaboratory tests which can be measurements, analyzes or experiments and which are carried out independently several times, have shown that the Analytik Jena solution provides specific and sensitive results.

The past and the future

Analyze wastewater samples biological parameters through the extraction of genetic information was something unthinkable just a few years ago, and not just because of technology and the efficiency of laboratory work.

Sampling from treatment plants is considered difficult. Silvio Beier, professor of technologies for Use of material flows in an urban environment to Bauhaus University of Weimarsaid:

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

Other applications for Legionella

Beier leads the Corona Monitoring in Thuringia (CoMoTH) project, which involves 23 sewage treatment plants covering around 40% of the population of the state of Thuringia, and has Analytik Jena as a participating partner. The goal is to explore opportunities for comprehensive and ongoing wastewater surveillance 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 within the population.”

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

Therefore, in addition to the monitoring advice for SARS-CoV-2, Analytik Jena already has an applications manual for Legionella and another for other relevant markers in wastewater, such as noroviruses and faecal markers.

(Photo credit: Analytik Jena).

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