US 11,674,124 B2
Methods for producing mutant bacteriophages for the detection of listeria
Jose S. Gil, Winnetka, CA (US); Dwight L Anderson, Minneapolis, MN (US); Stephen Erickson, White Bear Township, MN (US); and Minh Mindy Bao Nguyen, Shoreview, MN (US)
Assigned to Laboratory Corporation of America Holdings, Burlington, NC (US)
Filed by Laboratory Corporation of America Holdings, Burlington, NC (US)
Filed on Jun. 18, 2020, as Appl. No. 16/905,607.
Claims priority of provisional application 62/864,894, filed on Jun. 21, 2019.
Prior Publication US 2020/0399614 A1, Dec. 24, 2020
Int. Cl. A61K 31/7105 (2006.01); A61K 35/15 (2015.01); A61K 35/17 (2015.01); A61K 39/00 (2006.01); A61K 39/395 (2006.01); C12N 7/00 (2006.01); C12N 1/20 (2006.01); C12R 1/01 (2006.01)
CPC C12N 7/00 (2013.01) [C12N 1/205 (2021.05); C12N 2795/00021 (2013.01); C12N 2795/00051 (2013.01); C12R 2001/01 (2021.05)] 29 Claims
 
1. A method of producing a mutant phage with expanded host range, comprising:
(i) preparing a plurality of co-culture mixtures of varying ratios, comprising a host bacterial strain and a target-host bacterial strain;
(ii) adding a phage strain to each of the plurality of the co-culture mixtures;
(iii) incubating the plurality of the co-culture mixtures and the phage strain under bacterial culture conditions;
(iv) collecting a phage lysate from each of the plurality of the co-culture mixtures;
(v) pooling the collected phage lysates from each of the plurality of the co-culture mixtures;
(vi) assaying the pooled phage lysates to determine if host phage of the phage strain has expanded; and,
(vii) isolating the mutant phage with expanded host range,
wherein the mutant bacteriophage with expanded host range is a recombinant mutant bacteriophage with expanded host range comprising an indicator gene inserted into a late gene region of genome.