Portable device for rapid identification of locally sourced phage for use in phage therapy

S.V. Angus, B. Swan, C. Stewart, A.S. Cowan
Guild Associates, Inc.,
United States

Keywords: bacteriophage, portable, phage enrichment, sewage, lytic phage, phage lysate, multi drug-resistant organisms, PhageHD


Multi drug-resistant organisms (MDRO) are increasingly omnipresent in the healthcare settings of the public and military. Pathogens of interest in particular are those in the ESKAPEE group (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter spp, and E. coli spp). . As the prevalence of MDRO has increased, the number of new antibiotics to treat these infections has decreased. This has caused the rise of organisms resistant to all known antibiotics. To solve this, bacteriophage have been suggested and used as a treatment route for MDRO. Bacteriophage are highly specific and as a result, isolating them has historically been a months-long, labor intensive, and high-cost process. This was done in four steps: 1) purify the environmental sample; 2) concentrate resulting phage lysate; 3) combine lysate with target bacteria to enrich; and 4) purify screened phage lysate. This is done automatically with no user intervention required except adding and retrieving the sample. Currently, the system, PhageHD (Phage Hunting Device) has been evaluated with 250 – 500 mL raw sewage samples obtained from four locations across the US covering the Pacific Northwest, Midwest, South Atlantic Coastal, and North Atlantic Coastal. There was no countable difference in phage concentration after purification of the environmental sample as determined by phage titration using top agar when compared to tradition polyethylene glycol precipitation and centrifugation methods. PhageHD is an easy to use, portable, self-contained, and rapid device for isolation of lytic phage cocktails from aqueous samples in remote or austere locales against user-selected permissive and target bacterial strains without user intervention. Additionally, the initial purification and concentration steps can be performed independently of the enrichment steps if the user wishes to rapidly collect multiple samples for transport and full susceptibility analysis. The increased availability of locally identified phages will allow phage therapy against more MDRO, phage detection of environmental bacteria, and diagnosis of difficult to diagnose, low concentration bacteria. This simple method to screen phage against bacteria will be usable by hospitals, military personnel, and public health professionals to combat MDRO.