Arise - Evaluation of innovative molecular analysis methods and approaches for monitoring antibiotic resistance in wastewater
Antimicrobial resistance (AMR) is one of the greatest medical challenges. It causes costs in the healthcare system and increasingly leads to fatal outcomes in infections that were previously considered easily treatable. Precise knowledge of epidemiological events is an important pillar in the fight against this global threat and requires coordinated measures at all levels.
To protect people and the environment, mandatory monitoring of AMR in wastewater is now to be introduced as part of the revision of the European Urban Wastewater Directive (91/271/EEC). However, unlike viral pathogens such as SARS-CoV-2 or polio, AMR wastewater monitoring is still in its infancy.
The aim of the ARISE project is to create the essential methodological basis and to develop, test and realise forward-looking analysis concepts for the monitoring of antibiotic resistance in wastewater. For the first time, 1) all essential molecular analysis methods for the detection of antibiotic resistance genes (ARG) will be subjected to a systematic and comparative evaluation, 2) the entire analysis workflow (from sampling technology, sample preparation and extraction to bioinformatic-biostatistical evaluation) will be evaluated and optimised using suitable process controls for wastewater monitoring, and 3) innovative analysis approaches for the universal microbiological comparability of the results of the ARG analysis methods will be tested.
Various methods are being evaluated, such as quantitative PCR, digital PCR, shotgun metagenome sequencing, array-based high-throughput qPCR and a new capture assay-based high-throughput sequencing method for the targeted enrichment and quantitative analysis of AMR markers. The analytical methods will be evaluated in a uniform and comparative manner under standardised laboratory test conditions and in selected case studies. In addition, a combination of molecular biological and classical culture-based methods will provide a complete picture of the comparability and informative value of the methods, which is not currently available in this form.
The comprehensive consideration of innovative molecular AMR analysis methods paired with a systematic comparative study of their performance characteristics and the novel combination with relevant accompanying diagnostics for a well-founded interpretability of the results leads in summary to a solid knowledge and data base that will enable the successful implementation of wastewater-based AMR monitoring in the future for those in need - as proposed by the EU.