초록

The present invention relates, in part, to methods and kits for rapidly determining antimicrobial susceptibility of microorganisms.

대표청구항 ▼

1. A method for determining antimicrobial susceptibility of microorganisms comprising: incubating a liquid suspension of microorganisms in the presence of an antimicrobial under conditions that promote growth of the microorganisms;adding a signaling agent that binds to a surface of the microorganism

1. A method for determining antimicrobial susceptibility of microorganisms comprising: incubating a liquid suspension of microorganisms in the presence of an antimicrobial under conditions that promote growth of the microorganisms;adding a signaling agent that binds to a surface of the microorganisms;separating the microorganisms bound by the signaling agent from unbound signaling agent; andmeasuring signal levels associated with the microorganisms as compared to one or more controls, thereby determining the antimicrobial susceptibility of the microorganisms;wherein the signaling agent comprises a structure that is: 2. The method of claim 1, wherein the antimicrobial susceptibility of the microorganisms is determined in less than 5 hours. 3. The method of claim 1, wherein adding the signaling agent occurs during the incubating step. 4. The method of claim 1, wherein adding the signaling agent occurs after the incubating step. 5. The method of claim 1, wherein the signaling agent forms a non-covalent bond with the surface of the microorganism. 6. The method of claim 1, wherein the signaling agent forms a covalent bond with the surface of a microorganism in the presence of one or more agents that promote coupling, selected from the group consisting of glutaraldehyde, formaldehyde, paraformaldehyde, EDC, DCC, CMC, DIC, HATU, Woodward's Reagent, N,N′-carbonyl diimidazole, acrylates, amides, imides, anhydrides, chlorotriazines, epoxides, isocyanates, isothiocyanates, organic acids, monomers, polymers, silanes, silcates, NHS, and sulfo-NHS, and a combination thereof. 7. The method of claim 6, wherein the agent that promotes coupling is glutaraldehyde. 8. The method of claim 1, wherein multiple antimicrobials are tested in parallel. 9. The method of claim 1, wherein the determining signal levels comprises measuring the signal levels associated with intact microorganisms. 10. The method of claim 9, wherein the method further comprises a step of determining whether a microorganism is resistant, intermediately resistant, or susceptible to one or more antimicrobials and/or determining one or more antimicrobial minimum inhibitory concentrations (MIC) based upon the signal levels associated with intact microorganisms. 11. The method of claim 1, wherein the microorganisms are bacteria, fungi, protozoa, and/or archaea. 12. The method of claim 1, wherein the method does not involve a step of capturing microorganisms on a solid surface prior to or during incubation and/or does not include a step of growing microorganisms on a solid surface prior to or subsequent to the incubating step. 13. The method of claim 1, wherein the separating the microorganisms is performed by centrifugation, magnetic separation, filtration, electrophoresis, dielectrophoresis, precipitation, or agglutination, or a combination thereof. 14. The method of claim 1, wherein the one or more controls comprise a positive control measured from microorganisms under otherwise identical conditions but without antimicrobials or with one or more antimicrobials for which the microorganisms are not susceptible. 15. The method of claim 1, wherein the microorganisms are obtained from a biological sample from a subject having an infection of the microorganisms and/or are obtained from a culture derived from the biological sample. 16. The method of claim 15, wherein the biological sample is selected from the group consisting of blood or blood components, bronchoalveolar lavage, cerebrospinal fluid, nasal swabs, sputum, stool, throat swabs, vaginal swabs, urine, and wound swabs, or a combination thereof. 17. The method of claim 1, wherein the steps of incubating the liquid suspension of microorganisms and adding the signaling agent occur in a cartridge comprising a plurality of chambers and the step of determining signal levels associated with the microorganisms comprises determining signaling levels in the plurality of chambers. 18. The method of claim 17, wherein the cartridge further comprises one or more control chambers that do not contain antimicrobials or one or more antimicrobials for which the microorganisms are not susceptible. 19. A method for determining antimicrobial susceptibility of microorganisms comprising: incubating a liquid suspension of microorganisms in the presence of an antimicrobial and a signaling agent under conditions that promote growth of the microorganisms, wherein the signaling agent binds to a surface of the microorganisms;separating the microorganisms bound by the signaling agent from the unbound signaling agent; andmeasuring signal levels associated with the microorganisms as compared to one or more controls, thereby determining the antimicrobial susceptibility of the microorganisms;wherein the signaling agent comprises a structure that is: 20. The method of claim 19, wherein the antimicrobial susceptibility of the microorganisms is determined in less than 5 hours. 21. The method of claim 19, wherein the one or more controls comprise a positive control measured from microorganisms under otherwise identical conditions but without antimicrobials or with one or more antimicrobials for which the microorganisms are not susceptible. 22. The method of claim 19, wherein the signaling agent forms a non-covalent bond with the surface of a microorganism. 23. The method of claim 19, wherein the signaling agent forms a covalent bond with the surface of a microorganism in the presence of one or more agents that promote coupling, selected from the group consisting of glutaraldehyde, formaldehyde, paraformaldehyde, EDC, DCC, CMC, DIC, HATU, Woodward's Reagent, N,N′-carbonyl diimidazole, acrylates, amides, imides, anhydrides, chlorotriazines, epoxides, isocyanates, isothiocyanates, organic acids, monomers, polymers, silanes, silcates, NHS, and sulfo-NHS, and a combination thereof. 24. The method of claim 23, wherein the agent that promotes coupling is glutaraldehyde. 25. The method of claim 19, wherein the microorganisms are obtained from a biological sample from a subject having an infection of the microorganisms and/or are obtained from a culture derived from the biological sample; and wherein the biological sample is selected from the group consisting of blood or blood components, bronchoalveolar lavage, cerebrospinal fluid, nasal swabs, sputum, stool, throat swabs, vaginal swabs, urine, and wound swabs, or a combination thereof. 26. A method for determining antimicrobial susceptibility of microorganisms comprising: incubating a liquid suspension of microorganisms in a cartridge comprising a plurality of chambers, each chamber containing one or more antimicrobials, under conditions that promote growth of the microorganisms;adding a signaling agent to the plurality of chambers, wherein the signaling agent binds to a surface of the microorganisms;removing unbound signaling agent; andmeasuring signaling levels in the plurality of chambers as compared to one or more controls, thereby determining the susceptibility of microorganisms to the one or more antimicrobials;wherein the signaling agent comprises a structure that is: 27. The method of claim 26, wherein the antimicrobial susceptibility of the microorganisms is determined in less than 5 hours. 28. The method of claim 26, wherein the cartridge further comprises one or more control chambers that do not contain antimicrobials or one or more antimicrobials for which the microorganisms are not susceptible. 29. The method of claim 26, wherein the signaling agent forms a covalent bond with the surface of a microorganism in the presence of one or more agents that promote coupling, selected from the group consisting of glutaraldehyde, formaldehyde, paraformaldehyde, EDC, DCC, CMC, DIC, HATU, Woodward's Reagent, N,N′-carbonyl diimidazole, acrylates, amides, imides, anhydrides, chlorotriazines, epoxides, isocyanates, isothiocyanates, organic acids, monomers, polymers, silanes, silcates, NHS, and sulfo-NHS, and a combination thereof. 30. The method of claim 26, wherein the microorganisms are obtained from a biological sample from a subject having an infection of the microorganisms and/or are obtained from a culture derived from the biological sample; and wherein the biological sample is selected from the group consisting of blood or blood components, bronchoalveolar lavage, cerebrospinal fluid, nasal swabs, sputum, stool, throat swabs, vaginal swabs, urine, and wound swabs, or a combination thereof.