Reporter plasmid phage packaging system for detection of bacteria

A method for producing transducing particles that comprise a bacteriophage coat and a DNA core that comprises plasmid DNA, said method comprising:

• a) introducing plasmid DNA into a bacterial host, wherein the plasmid DNA comprises
• i) a host-specific bacteriophage packaging site wherein the packaging site is substantially in isolation firom sequences naturally occurring adjacent thereto in the bacteriophage genome,
• ii) a reporter gene,
• iii) a bacteria-specific promoter operably linked to said reporter gene,
• iv) a bacteria-specific origin of replication, and optionally
• v) an antibiotic resistance gene,
• b) infecting the bacterial host with a host-specific bacteriophage, and
• c) collecting progeny phage transducing particles that comprise said plasmid DNA.

A method of detecting target bacteria present in a biological sample, said method comprising:

• a) exposing a biological sample to a transducing particle, wherein said transducing particle comprises a bacteriophage coat and a DNA core that comprises plasmid DNA, said plasmid DNA comprising:
• i) a host-specific bacteriophage packaging site wherein the packaging site is substantially in isolation from sequences naturally occurring adjacent thereto in the bacteriophage genome,
• ii) a reporter gene,
• iii) a bacteria-specific promoter operably linked to said reporter gene,
• iv) a bacteria-specific origin of replication, and optionally
• v) an antibiotic resistance gene,
• b) incubating said bacterial sample under conditions to allow delivery of said plasmid DNA into the bacteria, and
• c) monitoring expression of said reporter gene comprised within said plasmid DNA, thereby detecting target bacteria present in the biological sample.