Rapid Cell Lysis for Molecular Environmental Diagnostics

Molecular diagnostics of health-relevant microorganisms and viruses is of vital interest for the protection of natur resources. The extraction of the DNA/RNA of pathogens and indicators from environmental samples is a complex, time-consuming process, requiring several hours and steps. After extraction DNA/RNA can be detected using amplification techniques such as PCR. As a consequence, state-of-the-art molecular diagnostics require well-equipped
laboratories and highly qualified personnel. In environmental diagnostics on-site test would be highly useful to investigate microbiological quality directly in the field and allow quick responses to deteriorating conditions. Rapid on-site testing would also be highly useful for quality control in the production and processing food or feed. The central goal of this project is to address one of the major bottlenecks in rapid molecular diagnostics: the lysis of target cells/virus particles and isolation of DNA/RNA in a simple, rapid and efficient manner. Cell lysis can be performed by hydrophilic ionic liquids (ILs) that can disrupt bacterial cells and stabilize the extracted DNA. This concept will be extended to other health-relevant targets: bacterial pathogens, enveloped and non-enveloped viruses, pathogenic fungi, and complex samples (water, faeces, soil). The objective is to select non-toxic and environmentally benign ILs that can
efficiently lyse the respective target and are compatible with downstream diagnostics. Furthermore, the various optimized cell lysis procedures will be extended to full-fledged DNA/RNA extraction methods. The goal is to develop
more rapid, simpler extraction procedures with DNA/RNA quality and yield comparable to state-of-the-art techniques. In a field trial a rapid field test kit for testing microbiological water quality will be developed and then evaluated by the Austrian Armed Forces in the course of exercises and deployments for emergency drinking water production in disaster settings.