ESCMID Global 2025

The RPA method requires minimal technical expertise and equipment, making it accessible for use in various settings. The robustness and efficiency of the RPA reaction is advantageous not only in the molecular diagnostics market, addressing the growing demand for rapid, reliable, and cost-effective DNA amplification methods, but also as an alternative to PCR in NGS library preparation steps.

Recombinase Polymerase Amplification (RPA) is a novel isothermal DNA amplification technique that operates at a constant low temperature (37-45 °C) with only two specific primers, offering a rapid and sensitive alternative to traditional temperature cycling requiring methods like PCR. High reaction temperatures not only require complex equipment but can damage fragile, scarce, fragmented samples. Therefore, RPA is a cost, energy, and sample saving option for nucleic acid amplification. It can be adapted in resource-limited settings and decentralized diagnostics applications.  

We developed RPA reagents compatible with lyophilization. Additionally, the RPA reaction conditions were optimized and the composition of the RPA reaction buffer adapted to ensure consistent and reliable results across diverse sample types. Moreover, we demonstrated the use of RPA reaction as a low temperature NGS library preamplification option.

Emerging research highlights the critical role of vaginal microbiome health in reproductive outcomes. Dysbiosis and other microbiological conditions of the female genitourinary system have been strongly linked to poor fertility outcomes. However, routine infertility care is hindered by the lack of standardized tools for microbiome characterization. Current standard of care tests are not suitable for microbiome profiling, and common approaches for microbiome analysis, such as sequencing, face significant technical limitations when applied to vaginal specimens. 

This presentation examines the technical barriers to vaginal microbiome analysis. It introduces PMP™, a precision microbiome profiling method utilizing highly parallelized single-plex quantitative PCR, demonstrates its performance for vaginal specimens and discusses its future potential for routine use in infertility care.

Wastewater serves as a mirror of society, offering valuable insights into a wide range of parameters. Molecular biology leverages this potential by regular analysis of wastewater samples to monitor the infection dynamics of various pathogens. In Vienna, respiratory viruses such as SARS-CoV-2, influenza, and RSV have been an integral part of wastewater monitoring for several seasons. Beyond viruses, antibiotic resistance genes of bacteria are of particular interest, as their spread is associated with a major public health threat. In this presentation, the above-mentioned possibilities of molecular biological wastewater monitoring, which are carried out at TU Wien, will be highlighted, and it will be discussed how quantitative analysis of different targets using dPCR can support public health efforts by providing precise and reliable data for infection surveillance and antimicrobial resistance tracking.