Methods for Generation of Reporter Phages and Immobilization of Active Bacteriophages on a Polymer Surface

Abstract

Novel reporter bacteriophages are provided. Provided are compositions and methods that allow bacteriophages that are used for specific detection or killing of E. coli 0157:H7 to be propagated in nonpathogenic E. coli , thereby eliminating the safety and security risks of propagation in E. coli 0157:H7. Provided are compositions and methods for attaching active bacteriophages to the surface of a polymer in order to kill target bacteria with which the phage comes into contact. Provided are modified bacteriophages immobilized to a surface, which capture E. coli 0157:H7 and cause the captured cells to emit light or fluorescence, allowing detection of the bacteria in a sample.

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         7 . A method for immobilizing a phage on a polymer surface, the method comprising:
 treating a polymer surface to increase the surface tension;   applying a mixture of monomer or oligomer, photoinitiator, and phage to the treated polymer surface; and   polymerizing the mixture to the polymer surface using UV light, thereby immobilizing the phage on the polymer surface.   
     
     
         8 . The method of  claim 7  wherein the phage is ΦV10. 
     
     
         9 . The method of  claim 8  wherein the immobilized phage was propagated in non-pathogenic  E. coli.    
     
     
         10 . The method of  claim 7  wherein the polymer surface is corona treated. 
     
     
         11 . The method of  claim 7  wherein the polymer surface is an inner surface of a stomacher bag. 
     
     
         12 . The method of  claim 7  wherein the immobilized phage is capable of capturing and infecting target bacteria. 
     
     
         13 . A polymer comprising a phage immobilized onto the polymer surface, the polymer obtained according to the method of  claim 7 . 
     
     
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