Methods of making recombinant phage, compositions and articles of manufacture of same for bacterial detection

• US 9,340,817 B2
• Filed: 03/27/2014
• Issued: 05/17/2016
• 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, Shivers, Robert Patrick, Brownell, Daniel Robert, Holder, Jason Wyatt, Bowers, Jayson Linn
• Primary Examiner(s): KINSEY WHITE, NICOLE ERIN
• Application Number:
  US14/226,889
• Publication Number:
  US 20140302487A1
• Time in Patent Office: 782 Days
• Field of Search: None
• US Class Current: 1/1
• 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 composition comprising A511, LP40, and LP124 recombinant Listeria phage, each recombinant phage comprising a heterologous nucleic acid sequence encoding a luciferase that is at least 70% identical to SEQ ID NO:

4, a β

-galactosidase, or a horseradish peroxidase.

3. The composition of claim 1, wherein said composition provides increased Listeria species host range detection in comparison to the host range of a single recombinant Listeria phage.

4. The composition of claim 1, wherein said composition provides increased sensitivity for the detection of Listeria species in comparison to the sensitivity of a single recombinant Listeria phage.

7. A method of detecting target bacteria, comprising:

exposing a sample to the composition of claim 1; and

assaying for the presence of the luciferase, β

-galactosidase or horseradish peroxidase in the exposed sample.

2. An article of manufacture comprising A511, LP40, and LP124 recombinant Listeria phage, each recombinant phage comprising a heterologous nucleic acid sequence encoding a luciferase that is at least 70% identical to SEQ ID NOs 4, a β

-galactosidase, or a horseradish peroxidase; 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 one compound selected from nitrogen containing compounds, c) at least one compound selected from nucleic acids and related compounds, d) at least one compound selected from lipid, e) at least one inorganic compound, and f) at least one organic compound.

5. The article of manufacture of claim 2, wherein said article of manufacture provides increased Listeria species host range detection in comparison to the host range of a single recombinant Listeria phage.

6. The article of manufacture of claim 2, wherein said article of manufacture provides increased sensitivity for the detection of Listeria species in comparison to the sensitivity of a single recombinant Listeria phage.

8. A method of detecting target bacteria, comprising:

exposing a sample to the article of manufacture of claim 2; and

assaying for the presence of the luciferase, β

-galactosidase or horseradish peroxidase in the exposed sample.