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1. WO2020117650 - APPARATUS, SYSTEMS, AND METHODS FOR QUANTIFYING INFECTIOUS AGENTS

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[ EN ]

CLAIMS

What is claimed is:

1. A method of determining a concentration of an infectious agent of an unknown strain in a target sample, the method comprising:

diluting a first aliquot of the target sample comprising the infectious agent of the unknown strain by a first dilution factor (DFi) to yield a first diluted sample; diluting a second aliquot of the target sample comprising the infectious agent of the unknown strain by a second dilution factor (DF2) to yield a second diluted sample;

determining a first time-to-detection (TTDi) representing the time it takes a

solution characteristic of the first diluted sample to undertake a predetermined threshold change;

determining a second time-to-detection (TTD2) representing the time it takes the solution characteristic of the second diluted sample to undertake the predetermined threshold change;

calculating an average calibration curve slope (maVg) and an average calibration curve y-intercept (bav ) from equation parameters obtained from multiple calibration curves representing growth behavior of one or more infectious agents of different known strains;

calculating a corrected calibration curve slope (mcorr) using at least the TTD2, the TTDi, the DF2, and the DFi;

calculating a corrected calibration curve y-intercept (bCOrr) using at least the bavg, the mcorr, and the mav ; and

determining the concentration of the infectious agent of the unknown strain in the target sample using at least the mcorr, the bcorr, and either the TTDi and the DFi or the TTD2 and the DF2.

2. The method of claim 1, wherein the one or more infectious agents of the different known strains comprise at least a first infectious agent and a second infectious agent, wherein the first infectious agent is a different species from the second infectious agent.

3. The method of claim 1, wherein the one or more infectious agents of the different known strains are the same species as the infectious agent of the unknown strain.

4. The method of claim 1, further comprising generating the multiple calibration curves prior to calculating the maVg and the baVg by:

preparing cultures comprising the one or more infectious agents of the different known strains, wherein the prepared cultures comprises different initial concentrations (Ninitiai) of an infectious agent of a known strain; monitoring, using one or more sensors, changes in the solution characteristics of each of the prepared cultures over time;

determining a calibration time-to-detection (TTDcaiibration) of each of the prepared cultures representing the time it takes the solution characteristic of each of the prepared cultures to undertake the predetermined threshold change; fitting each of the multiple calibration curves to TTDcaiibration data and Ninitiai data related to a specific known strain using the relationship:

TTDcaiibration = nistram_specific X loga(Ninitial) + bstrain_specific,

wherein a is any positive real number other than 1,

wherein mstrain_specific is a strain- specific calibration curve slope, and wherein bstrain_specific is a strain- specific calibration curve y-intercept.

5. The method of claim 4, wherein calculating the mav is taking an average of multiple mstrain_specific values and calculating the bavg is taking an average of multiple bstrain_specific values.

6. The method of claim 4, wherein calculating the mCOrr comprises using the relationship:


7. The method of claim 6, wherein calculating the bCOrr comprises using the relationship:


8. The method of claim 7, wherein determining the concentration of the infectious agent of the unknown strain (Conct^get) comprises using the relationship:

Cone target

9. The method of claim 7, wherein determining the concentration of the infectious agent of the unknown strain (Conctarget) comprises using the relationship:

TTD2 - bcorr

Conctarget = DF2 X a mcorr

10. The method of claim 1, wherein the first aliquot and the second aliquot of the sample are diluted with growth media.

11. The method of claim 1, wherein the solution characteristic is an oxidation reduction potential (ORP) and the solution characteristic is monitored by at least one computing device communicatively coupled to at least a first ORP sensor and a second ORP sensor, wherein each of the first ORP sensor and the second ORP sensor comprises a redox-active material, wherein the first ORP sensor is in fluid communication with the first diluted sample, and wherein the second ORP sensor is in fluid communication with the second diluted sample, wherein the ORP is monitored in the absence of any added reporter molecules in any of the first diluted sample or the second diluted sample.

12. The method of claim 11, wherein the first ORP sensor and the second ORP sensor each comprise at least an active electrode and a reference electrode.

13. The method of claim 11, wherein the predetermined threshold change is a change in the ORP of between approximately -100 mV and -700 mV.

14. The method of claim 11, wherein the redox-active material comprises a gold layer, a platinum layer, a metal oxide layer, a carbon layer, or a combination thereof.

15. The method of claim 1, wherein the solution characteristic is pH and the solution characteristic is monitored by at least one computing device communicatively coupled to at least a first pH sensor and a second pH sensor, wherein each of the first pH sensor and the second pH sensor comprise a functionalization layer, wherein the first pH sensor is in fluid communication with the first diluted sample, and wherein the second pH sensor is in fluid communication with the second diluted sample

16. The method of claim 15, wherein the first pH sensor and the second pH sensor each comprise at least an active electrode and a reference electrode.

17. The method of claim 15, wherein the predetermined threshold change is

approximately a change in pH of between approximately -0.01 to -3.0.

18. The method of claim 1, wherein the target sample comprises a bodily fluid, a wound swab or sample, a rectal swab or sample, another type of biological sample, a culture derived therefrom, or a combination thereof.

19. The method of claim 18, wherein the bodily fluid comprises urine, blood, sputum, saliva, breast milk, spinal fluid, semen, vaginal secretions, synovial fluid, pleural fluid, peritoneal fluid, pericardial fluid, amniotic fluid, cultures of bodily fluid that have tested positive for infectious agent growth, or a combination thereof.

20. The method of claim 1, wherein the infectious agent comprises bacteria, fungus, mold, or a combination thereof.

21. A system to determine a concentration of an infectious agent of an unknown strain in a target sample, comprising:

a metering conduit configured to:

dilute a first aliquot of the target sample comprising the infectious agent of the unknown strain by a first dilution factor (DFi) to yield a first diluted sample, and

dilute a second aliquot of the target sample comprising the infectious agent of the unknown strain by a second dilution factor (DF2) to yield a second diluted sample;

a first sensor configured to detect a change in a solution characteristic of the first diluted sample and a second sensor configured to detect a change in the solution characteristic of the second diluted sample;

one or more sample delivery conduits configured to introduce the first diluted

sample to the first sensor and introduce the second diluted sample to the second sensor;

a computing device communicatively coupled to the first sensor and the second sensor, wherein the computing device comprises one or more processors, wherein the one or more processors are programmed to:

determine a first time-to-detection (TTDi) representing the time it takes the solution characteristic of the first diluted sample to undertake a predetermined threshold change;

determine a second time-to-detection (TTD2) representing the time it takes the solution characteristic of the second diluted sample to undertake the predetermined threshold change;

calculate a corrected calibration curve slope (mcorr) using at least the TTD2, the TTDi, the DF2, and the DFi;

calculate an average calibration curve slope (maVg) and an average

calibration curve y-intercept (bav ) from equation parameters obtained from multiple calibration curves representing growth behavior of one or more infectious agents of different known strains

calculate a corrected calibration curve y-intercept (bcorr) using at least the mCorr, the maVg, and the bav ; and

determine the concentration of the infectious agent of the unknown strain in the target sample using at least the mcorr, the bcorr, and either the TTDi and the DFi or the TTD2 and the DF2.

22. The system of claim 21 wherein the one or more infectious agents of the different known strains comprise at least a first infectious agent and a second infectious agent, wherein the first infectious agent is a different species from the second infectious agent.

23. The system of claim 21 wherein the one or more infectious agents of the different known strains are the same species as the infectious agent of the unknown strain.

24. The system of claim 21 wherein the one or more processors are programmed to generate the multiple calibration curves prior to calculating the mav and the bav by:

monitor, via one or more sensors communicatively coupled to the computing

device, changes in the solution characteristics of prepared cultures comprising the one or more infectious agents of the different known strains, wherein the prepared cultures comprises different initial concentrations (Nmitial) of an infectious agent of a known strain;

determine a calibration time-to-detection (TTDcaiibration) of each of the prepared cultures representing the time it takes the solution characteristic of each of the prepared cultures to undertake the predetermined threshold change; fit each of the multiple calibration curves to TTDcaiibration data and Nmitial data related to a specific known strain using the relationship:

TTDcaiibration = nistram_specific X loga(Ninitial) + bstrain_specific, wherein a is any positive real number other than 1,

wherein mstrain_specific is a strain- specific calibration curve slope, and wherein bstrain_specific is a strain- specific calibration curve y-intercept.

25. The system of claim 24, wherein the one or more processors are programmed to calculate the maVg by taking an average of multiple mstrain_specific values and calculate the bavg by taking an average of multiple bstrain_spedfic values.

26. The system of claim 24, wherein the one or more processors are programmed to calculate the mcorr using the relationship:


27. The system of claim 26 wherein the one or more processors are programmed to calculate the bCOrr using the relationship:

h •Jcorr -— mcorr X L h Uavg- mavg

28. The system of claim 27, wherein the one or more processors are programmed to determine the concentration of the target infectious agent of the unknown strain (Conctarget) using the relationship:

Conctarget

29. The system of claim 27, wherein the one or more processors are programmed to determine the concentration of the target infectious agent of the unknown strain (Conctarget) using the relationship:

TTD2 - bcorr

Conctarget = DF2 X a mcorr

30. The system of claim 21, wherein the first aliquot and the second aliquot of the target sample are diluted with growth media.

31. The system of claim 21, wherein the solution characteristic is an oxidation reduction potential (ORP), wherein the first sensor and the second sensor are ORP sensors, and wherein each of the first sensor and the second sensor comprise a redox-active material.

32. The system of claim 31, wherein the predetermined threshold change is a change in the ORP of between approximately -100 mV and -700 mV.

33. The system of claim 31, wherein the redox-active material comprises a gold layer, a platinum layer, a metal oxide layer, a carbon layer, or a combination thereof.

34. The system of claim 21, wherein the solution characteristic is pH, wherein the first sensor and the second sensor are pH sensors, and wherein each of the first sensor and the second sensor comprise a functionalization layer.

35. The system of claim 34, wherein the predetermined threshold change is approximately a change in pH of between approximately -0.01 to -3.0.

36. The system of claim 21, wherein the first sensor and the second sensor each comprise at least an active electrode and a reference electrode.

37. The system of claim 21, wherein the target sample comprises a bodily fluid, a wound swab or sample, a rectal swab or sample, another type of biological sample, a sample culture derived therefrom, or a combination thereof.

38. The system of claim 37, wherein the bodily fluid comprises urine, blood, sputum, saliva, breast milk, spinal fluid, semen, vaginal secretions, synovial fluid, pleural fluid, peritoneal fluid, pericardial fluid, amniotic fluid, cultures of bodily fluid that have tested positive for infectious agent growth, or a combination thereof.

39. The system of claim 21, wherein the infectious agent comprises bacteria, fungus, mold, or a combination thereof.