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.
- 2. A method, as claimed in claim 1, wherein:
said step of making comprises assuming that any of said target bacterium that are present in said liquid solution are present in said liquid solution in a low concentration.
- 3. A method, as claimed in claim 1, wherein:
said step of making comprises performing an assay to determine if a biomarker for said target bacterium is present that is indicative of the presence of a concentration of said target bacterium that reliably exceeds the detection limit of the detection technology.
- 4. A method, as claimed in claim 3, wherein:
said step of performing an assay comprises performing a mass spectrum analysis.
- 5. A method, as claimed in claim 1, wherein:
said step of using comprises applying a first quantity of said bacteriophage to said liquid solution such that there is likely to be a high multiplicity of infection (“MOI”) number.
- 6. A method, as claimed in claim 1, wherein:
said step of analyzing comprises performing a mass spectrum analysis.
- 7. A method, as claimed in claim 1, wherein:
said step of analyzing comprises performing a MALDI analysis.
- 8. A method, as claimed in claim 1, wherein:
said step of analyzing comprises performing a MALDI-TOF analysis.
- 9. A method, as claimed in claim 1, wherein:
said step of analyzing comprises performing an electro-spray ionization mass spectrometry analysis.
- 10. A method, as claimed in claim 1, wherein:
said step of analyzing comprises performing an ion mobility spectrometry analysis.
- 11. A method, as claimed in claim 1, wherein:
said step of analyzing comprises performing an optical spectroscopy analysis.
- 12. A method, as claimed in claim 1, wherein:
said step of analyzing comprises performing an immuno analysis.
- 13. A method, as claimed in claim 1, wherein:
said step of analyzing comprises performing a chromatographic analysis.
- 14. A method, as claimed in claim 1, wherein:
said step of analyzing comprises performing an aptamer analysis.
- 15. A method, as claimed in claim 1, further comprising:
determining whether said liquid solution is likely to contain a biological element with a biomarker that could create a false positive for a biomarker for said bacteriophage.
- 16. A method, as claimed in claim 15, wherein:
said step of determining comprises assuming that a biological element is present in said liquid solution that has a biomarker that could create a false positive for a biomarker for said bacteriophage.
- 17. A method, as claimed in claim 15, wherein:
said step of determining comprises performing an assay to determine if a biomarker is present that could create a false positive for a biomarker for said bacteriophage.
- 18. A method, as claimed in claim 1 5, further comprising:
after a determination has been made that said biological element may present, separating said target bacterium from said liquid solution.
- 19. A method, as claimed in claim 1 8, wherein:
said step of separating comprises using immuno-magnet beads coated with antibodies for said target bacteria.
- 20. A method, as claimed in claim 1 8, further comprising:
after a determination has been made said step of separating is performed prior to said step of using.
- 21. A method, as claimed in claim 18, wherein:
after a determination has been made that said biological element may present and after said step of using, separating said bacteriophage from said liquid solution.
- 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.
- 23. A method, as claimed in claim 22, wherein:
said step of using comprises applying a first quantity of said biotinylated bacteriophage to said liquid solution such that there is likely to be a high multiplicity of infection (“MOI”) number.
- 24. A method, as claimed in claim 22, wherein:
said step of using comprises applying a first quantity of said biotinylated bacteriophage to said liquid solution that is near or likely to exceed the multiplicity of infection (“MOI”) number, wherein said first quantity of biotinylated bacteriophage are attached to a strepavidin coated probe.
- 25. A method, as claimed in claim 22, wherein:
said step of analyzing comprises separating a substantial portion of said biotinylated bacteriophage from said liquid solution to produce a remaining liquid solution.
- 26. A method, as claimed in claim 25, wherein:
said step of separating comprises using a strepavidin coated probe.
- 27. A method, as claimed in claim 26, wherein:
said step of using a strepavidin coated probe comprises separating a strepavidin coated probe from said liquid solution, said strepavidin coated probe having been used to apply said biotinylated bacteriophage to said liquid solution in said step of using.
- 28. A method, as claimed in claim 26, wherein:
said step of using a strepavidin coated probe comprises applying a strepavidin coated probe to said liquid solution to capture biotinylated bacteriophage in said liquid solution.
- 29. A method, as claimed in claim 25, wherein:
said step of analyzing comprises assaying said remaining liquid solution to determine if a biomarker for said bacteriophage is present.
- 30. A method, as claimed in claim 29, wherein:
said step of assaying comprises performing a mass spectrum analysis.
- 31. A method, as claimed in claim 22, further comprises:
separating said target bacterium from said mixture.
- 32. A method, as claimed in claim 31, wherein:
said step of separating is performed prior to said step of using.
- 33. A method, as claimed in claim 22, further comprises:
separating said biotinylated bacteriophage from said mixture.
- 34. A method, as claimed in claim 33, wherein:
said step of separating is performed after said step of using.
- 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.
- 36. A method, as claimed in claim 35, wherein:
said step of second using comprises performing a purification technique that separates at least some of any target bacterium that are present in said liquid solution from said liquid solution.
- 37. A method, as claimed in claim 35, wherein:
said step of performing occurs before said step of first using.
- 38. A method, as claimed in claim 37, wherein:
said step of performing comprises using immuno-magnetic beads to which are attached antibodies for said target bacterium.
- 39. A method, as claimed in claim 35, wherein:
said step of second using comprises performing a purification technique that separates said bacteriophage from said liquid mixture.
- 40. A method, as claimed in claim 39, wherein:
said step of performing occurs after said step of first using.
- 41. A method, as claimed in claim 39, wherein:
said step of performing comprises using immuno-magnetic beads to which are attached antibodies for said bacteriophage.
- 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.
- 43. A method, as claimed in claim 42, wherein:
said step of analyzing comprises separating said tagged bacteriophage from said liquid solution.
- 44. A method, as claimed in claim 42, wherein:
said step of analyzing comprises using a mass spectrometry analysis technique to produce a mass spectrum.
- 45. A method, as claimed in claim 44, wherein:
said step of using comprises inspecting said mass spectrum to determine if a biomarker that is unique to said untagged bacteriophage relative to said tagged bacteriophage is present.
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional Application No. 60/319,184, entitled “METHOD OF DETECTING LOW CONCENTRATIONS OF A TARGET BACTERIA THAT USES PHAGES TO INFECT TARGET BACTERIAL CELLS” and filed by Angelo J. Madonna and Kent J. Voorhees on Apr. 12, 2002, which application is incorporated by reference into this application in its entirety.
Provisional Applications (1)
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Number |
Date |
Country |
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60319184 |
Apr 2002 |
US |