Phage RealTime Amplification for Molecular Enumeration

Because of the increasing resistance of bacteria to antibiotics, research into other antibacterial therapies is crucial. Phage therapy plays an important role in this context. Phages are viruses that infect bacteria in a very specific way. In fact, most phages infect only a single bacterial species and usually only a few strains within that species. An increasing number of physicians and researchers recognize the potential of phage treatment in the control of bacterial infections. Within this framework, interventional studies are underway and numerous more studies are planned in which phages are administered to patients for the treatment of specific bacterial infections. Within these studies, methods are needed to quickly and accurately determine the change in bacterial and viral load to provide qualitative and quantitative monitoring of both the progression of the infection and the therapeutic agent, i.e., the phages. Currently, phage counts are usually determined by culture-based titration methods, a labor-intensive method with limited accuracy. In this project, we aim to develop a method to determine phage load based on real-time PCR or quantitative PCR (qPCR). To this end, we will focus on a) optimising the DNA extraction of phages, as no commercial kits are currently available for this purpose, and b) on the optimisation of qPCR formats for the 5 phages present in the BacterioFaag2 cocktail developed at the Queen Astrid Military Hospital for the treatment of major burn injuries infections caused by the bacteria Acinetobacter baumanii, Pseudomonas aeruginosa and Staphylococcus aureus.