US 11,952,611 B2
Methods and systems for detection of antibiotic resistance
Minh Mindy Bao Nguyen, Shoreview, MN (US); Dwight L. Anderson, Minneapolis, MN (US); Jose S. Gil, Winnetka, CA (US); Stephen E. Erickson, White Bear Township, MN (US); and Matthew J. Brown, Burlington, NC (US)
Assigned to Laboratory Corporation of America Holdings, Burlington, NC (US)
Filed by Laboratory Corporation of America Holdings, Burlington, NC (US)
Filed on Jul. 3, 2018, as Appl. No. 16/026,462.
Claims priority of provisional application 62/529,689, filed on Jul. 7, 2017.
Prior Publication US 2019/0010534 A1, Jan. 10, 2019
Int. Cl. C12Q 1/14 (2006.01); C12Q 1/06 (2006.01); C12Q 1/10 (2006.01); C12Q 1/18 (2006.01); C12Q 1/66 (2006.01); G01N 33/569 (2006.01); C12N 15/73 (2006.01); C12Q 1/689 (2018.01)
CPC C12Q 1/14 (2013.01) [C12Q 1/06 (2013.01); C12Q 1/10 (2013.01); C12Q 1/18 (2013.01); C12Q 1/66 (2013.01); G01N 33/56911 (2013.01); C12N 15/73 (2013.01); C12Q 1/689 (2013.01)] 21 Claims
 
1. A method of detecting antibiotic-resistant bacteria in a sample comprising:
(a) contacting the sample with an antibiotic,
(b) contacting the sample with a recombinant bacteriophage specific to the bacteria, wherein the recombinant bacteriophage comprises a genetic construct inserted into a late gene region of the bacteriophage genome, the genetic construct comprising:
(i) an indicator gene, wherein the indicator gene is not contiguous with a gene encoding a structural bacteriophage protein and does not yield a fusion protein; and
(ii) a promoter controlling transcription of the indicator gene, wherein the promoter is a bacteriophage late promoter,
and wherein expression of the indicator gene results in an indicator protein product, and wherein the indicator protein product is a luciferase;
(c) detecting a signal produced by the indicator protein product,
wherein detection of the signal is used to determine the presence of antibiotic-resistant bacteria in the sample.