Bacteriophage (phage) immobilization on substrates is important for many biocontrol and pathogen detection applications. We propose using electrospun biopolymer (polyhydroxybutyrate, (PHB)) nanofibers as substrate due to their large specific surface area and biocompatibility. Electrospun PHB fibers were fabricated for immobilization of a reporter phage for the detection of Escherichia coli O157: H7. In this study, the E. coli O157:H7-specific lytic phage PP01 was genetically engineered to express luceriferase, providing a bioluminescence signal upon detection of its host. The phage was immobilized through physical attachment by immersing electrospun fibers in PP01 suspensions. The surface morphology of the fiber was studied using scanning electron microscopy (SEM) and water contact angle (WCA) measurements. In addition, phage-staining and fluorescence microscopy were employed to investigate phage binding on the polymeric fiber substrate. We have shown that immobilized phages on electrospun fibers displayed higher infectivity than that of phages on plasma treated flat film. The infectivity of attached phages was confirmed through plaque and infection dynamic assays. We then demonstrated that the immobilized phages can be used to detect E.coli cells at a concentration of 102 CFU/ml within 4 hours. This work presents potential use of phages as biosensor and antimicrobial packaging.

Immobilization of reporter bacteriophage PP01 on electrospun PHB fibers for Escherichia coli O157: H7 detection

2019 Fall National Meeting

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