Poster abstracts
Poster number 7 submitted by Alfonso Carrillo
Capturing phage-host pairs with viral tag and grow
Alfonso Carrillo (The Ohio State University), Ami Fofana (The Ohio State University), Courtney M. Sanderson (The Ohio State University), Marissa R. Gittrich (The Ohio State University), Matthew B. Sullivan (The Ohio State University), Ger Van Den Engh (Center for Marine Cytometry)
Abstract:
Bacteriophages (phages) are viruses that exclusively infect bacteria and have long been considered as a viable alternative/supplement to antibiotic treatment due to their ability to lyse bacteria with minimal impact on the bacterial host. Continued advances in culture-independent sequenced-based analyses have allowed for viral discovery to advance at an astonishing rate. However, without isolating viruses these metagenomic approaches are unable to immediately associate viruses with their hosts and answer the fundamental question: who infects whom? Here we develop a scalable approach to link phages to their host cells using flow cytometric viral tagging followed by characterization of whether phage-host pairs reflect adsorption or successful lytic infections using a growth assay. This approach was challenged with the phage phiKZ and its host Pseudomonas aeruginosa PAO1M. We observed fluorescent shifts within our samples consisting of nucleic acid stained phage and unstained bacteria that indicate adsorption of the phage to the host. Observed shifts in fluorescence were directly correlated with changes in the phage concentration within the sample. Characterization of the infection through a growth assay was unsuccessful due to inaccurate sorting because of defective charging plates on the machine. This data supports previous findings that adsorption can be detected using flow cytometry, while characterization of the infection through a sorted growth assay requires further development. Further investigation of the ability to sort single phage-host pairs and characterize infection through a growth assay may illustrate the ability for this method to become a scalable approach for phage-host discovery of lytic phages. These findings provide the foundation for a scalable approach to phage-host discovery that can be used for library construction and exploring ecological population structure.
References:
Jang, H.B., Chittick, L., Li, YF. et al. Viral tag and grow: a scalable approach to capture and characterize infectious virus–host pairs. ISME COMMUN. 2, 12 (2022). https://doi.org/10.1038/s43705-022-00093-9
Keywords: Flow Cytometry, Viral Tagging, Phages