Performing antimicrobial susceptibility testing and related systems and methods

1. An automated rapid antimicrobial susceptibility testing system for performing a multi-assay testing sequence, the system comprising:

one or more automated incubation assemblies comprising a nest assembly adapted to: i) house at least one test panel having a plurality of wells for receiving a sample comprising microorganisms, and ii) facilitate incubation of one or more test panels in order to undergo the multi-assay testing sequence:

one or more agitation systems configured to generate a repeated motion of the nest assemblies;

a centrifugation system configured to separate microorganisms from a remainder of one or more samples or diluents within the plurality of wells of the test panel by applying a relative centrifugal force of about 2000 g to about 5000 g to the test panel, thereby forming a pellet of the microorganisms within each of the plurality of wells of the test panel;

a robotic handling assembly configured to accept one or more incoming test panels and move them between two assemblies of the system and/or between an assembly and the centrifugation system between steps of the multi-assay testing sequence;

an automated liquid handling assembly configured to exchange one or more fluids in the plurality of wells of the test panels, wherein the liquid handling assembly comprises at least one liquid addition system and an aspiration system and wherein the aspiration system is configured to remove fluid from a portion of a well of the test panel that is away from the pellet formed by the centrifugation system; and

an optical assembly for interrogation and readout of each assay of the multi-assay testing sequence being performed in the plurality of wells.

2. The system of claim 1 , wherein:

at least one nest assembly comprises a tiered frame comprising one or more floors, each floor comprising:

a stage to accommodate a test panel;

one or more cartridge positioning features extending from the stage; and

a set of recesses to accommodate a test panel handling device; and

the agitation system is configured to generate a repeated motion of the tiered frame.

3. The system of claim 2 wherein the tiered frame comprises multiple floors where each floor comprises at least one surface to accommodate at least one test panel.

4. The system of claim 2 wherein each floor comprises a heating element disposed in or along the stage.

5. The system of claim 1 wherein incubation system assembly is heated using forced air.

6. The system of claim 1 wherein the agitation system comprises a rotational agitation system comprising a rotating oscillating component.

7. The system of claim 6 wherein the agitation system comprises one or more linear bearing surfaces.

8. The system of claim 7 wherein the agitation system comprises two linear bearing surfaces positioned substantially perpendicularly with respect to one another and the agitation system comprises two sliding stages configured to slide along the bearing rails.

9. The system of claim 8 wherein the agitation system is configured to agitate the nest assembly along an orbital path having a radius that is about 1 mm to about 12 mm.

10. The system of claim 1 wherein the agitation system is configured to agitate the nest assembly along an orbital path and to vary a radius of the orbital path of agitation.

11. The system of claim 1 wherein the agitation system is configured to agitate the nest assembly along an orbital path at a rate of about 50 revolutions per minute to about 650 revolutions per minute.

12. The system of claim 1 wherein the robotic handling assembly is configured to move test panels between at least one of the one or more automated incubation assemblies and the optical assembly.

13. The system of claim 1 wherein the robotic handling assembly is configured to move test panels between at least one of the one or more automated incubation assemblies and an area accessible by the automated liquid handling assembly.

14. The system of claim 1 wherein reagents are stored in a disposable container.

15. The system of claim 1 wherein the optical assembly is configured to measure at least one of absorbance, fluorescence, luminescence, time-resolved fluorescence, or time-gated luminescence emitted from the sample during the multi-assay testing sequence.

16. The system of claim 15 wherein an excitation wavelength to generate a fluorescence emission is from about 545 nm to about 575 nm and a wavelength of the emission is from about 570 nm to about 610 nm.

17. The system of claim 15 wherein an excitation wavelength to generate a time-gated luminescence emission is about 280 nm to about 360 nm and a wavelength of the emission is one or more of the following: about 545 nm, about 565 nm, from about 608 nm to about 623 nm, or about 645 nm.

18. The system of claim 1 wherein the system is configured to yield a testing sequence throughput of at least 2, at least 4, at least 6, at least 8, at least 10, at least 12, at least 16, or at least 20 test panels per hour.

19. The system of claim 1 wherein a time duration for processing a test panel through the testing sequence from insertion of the test panel into the system to obtaining a result is less than 8 hours, less than 6 hours, less than 5 hours, less than 4 hours, less than 3 hours, or less than 2 hours.

20. The system of claim 1 wherein the fluid handling assembly comprises a manifold fluidly connected to at least one of the liquid addition system or the aspiration system, the manifold being configured to dispense and/or aspirate fluid using at least 8 tips.

21. The system of claim 1 , wherein the test panel comprises:

a first set of wells, comprising

a growth well receiving a nutrient broth and the sample; and

a no-growth well receiving the nutrient broth but not the sample; and

a second set of wells, comprising:

a plurality of antimicrobials, each antimicrobial applied to a plurality of wells in a dilution series.