Recombinant Phage and Bacterial Detection Methods

• US 20140302487A1
• Filed: 03/27/2014
• Published: 10/09/2014
• Est. Priority Date: 03/27/2013
• Status: Active Grant

• Current Assignee: Institute for Environmental Health, Inc.
• Original Assignee: Sample Technologies LLC
• Inventors: Koeris, Michael Sandor, Brownell, Daniel Robert, Shivers, Robert Patrick, Holder, Jason Wyatt, Bowers, Jayson Linn
• Granted Patent: US 9,340,817 B2
• Time in Patent Office: Days
• US Class Current: 435/5
• CPC Class Codes: C12N 2795/00031 Uses of virus other than therapeutic or vaccine, e.g. disinfectant
• CPC Class Codes: C12N 7/00 Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00
• CPC Class Codes: C12Q 1/04 Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor C12Q1/6897 takes precedence

Abstract

Methods of detecting target bacteria are provided. In some embodiments the methods comprise exposing the sample to a phage capable of infecting a set of target bacteria and comprising a heterologous nucleic acid sequence encoding a marker. In some embodiments the target bacteria comprise Listeria. In some embodiments the target bacteria are all Listeria. Recombinant Listeria phage comprising a heterologous nucleic acid sequence encoding a marker are also provided as are useful combinations of such phage and articles of manufacture comprising such phage, among other things.

Claims

1. A method of detecting target bacteria, comprising:

providing a sample;

exposing the sample to a first type of phage capable of infecting a first set of target bacteria and comprising a heterologous nucleic acid sequence encoding a first marker;

exposing the sample to a second type of phage capable of infecting a second set of target bacteria and comprising a heterologous nucleic acid sequence encoding a second marker; and

assaying for the presence of the first marker and the second marker in the exposed sample.

2. A method of detecting target bacteria, comprising:

providing a sample;

exposing the sample to at least one recombinant Listeria phage comprising a heterologous nucleic acid sequence encoding a marker, the recombinant Listeria phage selected from recombinant LP40 and derivatives thereof, recombinant LP48 and derivatives thereof, recombinant LP99 and derivatives thereof, recombinant LP101 and derivatives thereof, recombinant LP124 and derivatives thereof, recombinant LP125 and derivatives thereof, and recombinant LP143 and derivatives thereof; and

assaying for the presence of the marker in the exposed sample.

3. A recombinant Listeria phage comprising a heterologous nucleic acid sequence encoding a marker, the recombinant Listeria phage selected from recombinant LP40 and derivatives thereof, recombinant LP48 and derivatives thereof, recombinant LP99 and derivatives thereof, recombinant LP101 and derivatives thereof, recombinant LP124 and derivatives thereof, recombinant LP125 and derivatives thereof, and recombinant LP143 and derivatives thereof.

4. A recombinant Listeria phage comprising a heterologous nucleic acid sequence encoding a luciferase that is at least 70% identical to SEQ ID NO:

2, the recombinant Listeria phage selected from recombinant A511 and derivatives thereof, recombinant P100 and derivatives thereof, recombinant LP40 and derivatives thereof, recombinant LP48 and derivatives thereof, recombinant LP99 and derivatives thereof, recombinant LP101 and derivatives thereof, recombinant LP124 and derivatives thereof, recombinant LP125 and derivatives thereof, and recombinant LP143 and derivatives thereof.

5. A recombinant Listeria phage comprising a heterologous nucleic acid sequence encoding a luciferase at least 70% identical to SEQ ID NO:

4, the recombinant Listeria phage selected from recombinant A511 and derivatives thereof, recombinant P100 and derivatives thereof, recombinant LP40 and derivatives thereof, recombinant LP48 and derivatives thereof, recombinant LP99 and derivatives thereof, recombinant LP101 and derivatives thereof, recombinant LP124 and derivatives thereof, recombinant LP125 and derivatives thereof, and recombinant LP143 and derivatives thereof.

6. A composition comprising:

at least one recombinant Listeria phage comprising a heterologous nucleic acid sequence encoding a marker, the recombinant Listeria phage selected from recombinant LP40 and derivatives thereof, recombinant LP48 and derivatives thereof, recombinant LP99 and derivatives thereof, recombinant LP101 and derivatives thereof, recombinant LP124 and derivatives thereof, recombinant LP125 and derivatives thereof, and recombinant LP143 and derivatives thereof; and

at least one non-phage component selected from at least one of a) at least one compound selected from carbohydrates and related compounds, b) at least compound selected from nitrogen containing compounds, c) at least compound selected from nucleic acids and related compounds, d) at least compound selected from lipid, e) at least one inorganic compound, and f) at least one organic compound.

7. A composition comprising at least two recombinant Listeria phage comprising a heterologous nucleic acid sequence encoding a marker, the recombinant Listeria phage selected from recombinant A511 and derivatives thereof, recombinant P100 and derivatives thereof, recombinant LP40 and derivatives thereof, recombinant LP48 and derivatives thereof, recombinant LP99 and derivatives thereof, recombinant LP101 and derivatives thereof, recombinant LP124 and derivatives thereof, recombinant LP125 and derivatives thereof, and recombinant LP143 and derivatives thereof.

8. An article of manufacture comprising:

at least one recombinant Listeria phage comprising a heterologous nucleic acid sequence encoding a marker, the recombinant Listeria phage selected from recombinant LP40 and derivatives thereof, recombinant LP48 and derivatives thereof, recombinant LP99 and derivatives thereof, recombinant LP101 and derivatives thereof, recombinant LP124 and derivatives, recombinant LP125 and derivatives thereof, and recombinant LP143 and derivatives thereof; and

a solution comprising at least one non-phage component selected from at least one of a) at least one compound selected from carbohydrates and related compounds, b) at least compound selected from nitrogen containing compounds, c) at least compound selected from nucleic acids and related compounds, d) at least compound selected from lipid, e) at least one inorganic compound, and f) at least one organic compound.

9. An article of manufacture comprising at least two recombinant Listeria phage comprising a heterologous nucleic acid sequence encoding a marker, the recombinant Listeria phage selected from recombinant A511 and derivatives thereof, recombinant P100 and derivatives thereof, recombinant LP40 and derivatives thereof, recombinant LP48 and derivatives thereof, recombinant LP99 and derivatives thereof, recombinant LP101 and derivatives thereof, recombinant LP124 and derivatives thereof, recombinant LP125 and derivatives thereof, and recombinant LP143 and derivatives thereof;

and

a solution comprising at least one non-phage component selected from at least one of a) at least one compound selected from carbohydrates and related compounds, b) at least compound selected from nitrogen containing compounds, c) at least compound selected from nucleic acids and related compounds, d) at least compound selected from lipid, e) at least one inorganic compound, and f) at least one organic compound.

10. A method of identifying target bacteria of a phage, comprising:

exposing a plurality of liquid culture samples of different bacterial types to the phage; and

determining whether the phage infects the plurality of different bacterial types.