Sensitive and rapid determination of antimicrobial susceptibility
The present invention relates to moving microorganisms to a surface, where they are grown in the presence and absence of antimicrobials, and by monitoring the growth of the microorganisms over time in the two conditions, their susceptibility to the antimicrobials can be determined. The microorganisms can be moved to the surface through electrophoresis, centrifugation or filtration. When the movement involves electrophoresis, the presence of oxidizing and reducing reagents lowers the voltage at which electrophoretic force can be generated and allows a broader range of means by which the target can be detected. Monitoring can comprise optical detection, and most conveniently includes the detection of individual microorganisms. The microorganisms can be stained in order to give information about their response to antimicrobials.
1. A method of detecting growth of a microorganism in a sample comprising:
contacting a sample comprising a microorganism with a device comprising a chamber, wherein the chamber comprises a detection zone, and wherein the detection zone further comprises a hydrogel;
immobilizing the microorganism by capture with the hydrogel;
applying an electrophoretic force to the microorganism;
detecting, at a first time, a property of the microorganism with an optical detector;
detecting, at a second time, the property of the microorganism with the optical detector; and
determining the amount of difference of the property of the microorganism between the first time and the second time.
2. The method of claim 1 , wherein the hydrogel captures the microorganism by non-specific binding.
3. The method of claim 1 , wherein the electrophoretic force is applied continuously.
4. The method of claim 1 , wherein the electrophoretic force is applied intermittently.
5. The method of claim 4 , wherein the electrophoretic force is applied every 10 seconds.
6. The method of claim 1 , wherein the sample further comprises a growth medium.
7. The method of claim 6 , wherein the growth medium has a conductivity of less than 5 mS/cm.
8. The method of claim 6 , wherein the growth medium comprises a zwitterionic buffer.
9. The method of claim 6 , further comprising:
adding an anti-organism agent after determining the amount of difference of the property of the microorganism between the first time and the second time;
detecting, at a third time, the property of the microorganism with an optical detector;
detecting, at a fourth time, the property of the microorganism with the optical detector; and
determining the amount of difference of the property of the microorganism between the third time and the fourth time.