Method for Detecting Low Concentrations of a Target Bacterium That Uses Phages to Infect Target Bacterial Cells
First Claim
1. A method for determining if a target bacterium is present in a liquid solution when the target bacterium is or may be present in a low concentration that is near or below the detection limit for a particular detection technology comprising:
- making a determination as to whether a low concentration of a target bacterium may be present in a liquid solution;
using, following said step of making and if a determination is made that a low concentration of said target bacterium may be present, a first quantity of a bacteriophage to infect at least some of any target bacterium that are present in said liquid solution and multiply the number of bacteriophage in said liquid solution; and
analyzing, following said step of using, at least a portion of said liquid solution to determine if a biomarker for said bacteriophage is present that indirectly indicates that said target bacterium is also present in said liquid solution.
1 Assignment
0 Petitions
Accused Products
Abstract
The invention is directed to a method for detecting low concentrations of bacteria in liquid solution that may or may not be complex liquid solutions. In one embodiment, immunomagnetic separation (IMS) is used to separate target bacterium that may be in a liquid mixture from other constituents in the mixture. A low concentration of a bacteriophage for the target bacteria is subsequently used to infect target bacterial cells that have been captured using the IMS technique. If at least a certain concentration of target bacterium are present, the bacteriophage will multiply to a point that is detectable. Matrix assisted laser desorption ionization/time-of-flight-mass spectrometry (MALDI/TOF-MS) is then used to produce a mass spectrum that is analyzed to determine if one or more proteins associated with the bacteriophage are present, thereby indirectly indicating that target bacterium were present in the liquid mixture.
-
Citations
45 Claims
-
1. A method for determining if a target bacterium is present in a liquid solution when the target bacterium is or may be present in a low concentration that is near or below the detection limit for a particular detection technology comprising:
-
making a determination as to whether a low concentration of a target bacterium may be present in a liquid solution;
using, following said step of making and if a determination is made that a low concentration of said target bacterium may be present, a first quantity of a bacteriophage to infect at least some of any target bacterium that are present in said liquid solution and multiply the number of bacteriophage in said liquid solution; and
analyzing, following said step of using, at least a portion of said liquid solution to determine if a biomarker for said bacteriophage is present that indirectly indicates that said target bacterium is also present in said liquid solution. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21)
-
-
22. A method for determining if a target bacterium is present in a liquid solution when the target bacterium is or may be present in a low concentration that is near or below the detection limit for a particular detection technology comprising:
-
making a determination as to whether a low concentration of a target bacterium may be present in a liquid solution;
using, following said step of making and if a determination is made that a low concentration of said target bacterium may be present, a first quantity of a biotinylated bacteriophage to infect at least some of any target bacterium that are present in said liquid solution and multiply the number of bacteriophage in said liquid solution; and
analyzing, following said step of using, said liquid solution to determine if a biomarker for said bacteriophage is present that indirectly indicates that said target bacterium is also present in said liquid solution. - View Dependent Claims (23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34)
-
-
35. A method for determining if a target bacterium is present in a liquid solution when the target bacterium is or may be present in a low concentration that is near or below the detection limit for a particular detection technology comprising:
-
first determining whether a low concentration of a target bacterium may be present in a liquid solution;
second determining whether a biological element may be present in said liquid solution that has a biomarker that could create a false positive for a biomarker for a bacteriophage used in said subsequent step of using and used in said step of analyzing as an indirect biomarker for the presence of said target bacterium;
first using, following said step of first determining and when a determination has been made that a low concentration of said target bacterium may be present, a first quantity of a biotinylated bacteriophage to infect at least some of any target bacterium that are present in said liquid solution and multiply the number of bacteriophage in said liquid solution;
second using, following said step of second determining and when a determination has been made that a biological element may be present, purification technique; and
analyzing, following said steps of first using and second using, said liquid solution to determine if a biomarker for said bacteriophage is present that indirectly indicates that said target bacterium is also present in said liquid solution. - View Dependent Claims (36, 37, 38, 39, 40, 41)
-
-
42. A method for determining if a target bacterium is present in a liquid solution when the target bacterium is or may be present in a low concentration that is near or below the detection limit for a particular detection technology comprising:
-
making a determination as to whether a low concentration of a target bacterium may be present in a liquid solution;
using, following said step of making and if a determination is made that a low concentration of said target bacterium may be present, a first quantity of a tagged bacteriophage to infect at least some of any target bacterium that are present in said liquid solution and produce a number of untagged bacteriophage in said liquid solution; and
analyzing said liquid solution to determine if a biomarker for said untagged bacteriophage and that is distinguishable relative to any biomarkers for said tagged bacteriophage is present and that indirectly indicates that said target bacterium is also present in said liquid solution. - View Dependent Claims (43, 44, 45)
-
Specification