High throughput platform technology for rapid target identification in personalized phage therapy.
Fereshteh BayatArwa HilalMathura ThirugnanasampantharDenise TremblayCarlos D M FilipeSylvain MoinauTohid F DidarZeinab Hosseini-DoustPublished in: Nature communications (2024)
As bacteriophages continue to gain regulatory approval for personalized human therapy against antibiotic-resistant infections, there is a need for transformative technologies for rapid target identification through multiple, large, decentralized therapeutic phages biobanks. Here, we design a high throughput phage screening platform comprised of a portable library of individual shelf-stable, ready-to-use phages, in all-inclusive solid tablets. Each tablet encapsulates one phage along with luciferin and luciferase enzyme stabilized in a sugar matrix comprised of pullulan and trehalose capable of directly detecting phage-mediated adenosine triphosphate (ATP) release through ATP bioluminescence reaction upon bacterial cell burst. The tablet composition also enhances desiccation tolerance of all components, which should allow easier and cheaper international transportation of phages and as a result, increased accessibility to therapeutic phages. We demonstrate high throughput screening by identifying target phages for select multidrug-resistant clinical isolates of Pseudomonas aeruginosa, Salmonella enterica, Escherichia coli, and Staphylococcus aureus with targets identified within 30-120 min.
Keyphrases
- high throughput
- pseudomonas aeruginosa
- biofilm formation
- single cell
- acinetobacter baumannii
- cystic fibrosis
- multidrug resistant
- escherichia coli
- staphylococcus aureus
- endothelial cells
- stem cells
- transcription factor
- klebsiella pneumoniae
- high frequency
- loop mediated isothermal amplification
- gram negative
- induced pluripotent stem cells
- mesenchymal stem cells
- methicillin resistant staphylococcus aureus
- drug administration