Protocols for bacterial isolation are established but require a high load of bacteria and success rate varies. Axenic cultivation provides host cell-free genomic DNA for WGS, but ability to grow in axenic media varies between genotypes6. This WP will optimise current protocols for isolation of C. burnetii from positive field samples (WP1) by reducing contaminants and enriching bacterial load prior to cell culture and axenic inoculation. Therefore, decontamination procedures of samples will be evaluated, and cell culture systems optimized for C. burnetii recovery. Axenic media composition will be adapted to the requirements of different C. burnetii genotypes.
Additionally, immunomagnetic separation-based enrichment of bacteria from samples will be evaluated. The enhanced isolation protocol will be evaluated in Inter-Laboratory Proficiency Tests by project partners and implemented to obtain strains for WGS (WP3) and phenotypic and genotypic correlation (WP4). This WP will also generate a panel of C. burnetii reference strains representing all genotypes and from different host species which will be distributed to project partners for standardisation of surveillance protocols beyond the end of this project.
WP2 leads
Dr. Katja Mertens-Scholz
Katja is the head of the National Reference Laboratory for Q-Fever at the Friedrich-Loeffler Institute, Germany. Her work focuses on surveillance and molecular characterization of Coxiella burnetii.
Dr. Marcella Mori
Marcella is a molecular microbiologist and heads the Bacterial Zoonoses Unit at Sciensano, Brussels. She directs two National Reference Centres (Coxiella burnetii and Brucella spp.) for human health and multiple National Reference Laboratories for animals. Her research interests include enhancing animal diagnostics for C. burnetii and studying immune responses to improve disease control strategies.
