SYSTEMS AND METHODS FOR PRESUMPTIVE IDENTIFICATION OF MICROORGANISM TYPE IN A CULTURE

• US 20110208432A1
• Filed: 02/19/2008
• Published: 08/25/2011
• Est. Priority Date: 02/19/2008
• Status: Abandoned Application

• Current Assignee: Becton, Dickinson & Co
• Original Assignee: Becton, Dickinson & Co
• Inventors: Beaty, Patrick Shawn
• Application Number:
  US12/918,011
• Publication Number:
  US 20110208432A1
• Time in Patent Office: Days
• US Class Current: 702/19
• CPC Class Codes: C12M 41/46 of cellular or enzymatic activity or functionality, e.g. cell viability
• CPC Class Codes: C12Q 1/04 Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor C12Q1/6897 takes precedence

Abstract

Systems, methods and apparatus for identifying a microorganism type in a culture are provided. A normalization relative value is calculated for each respective measurement of a biological state of the culture between (i) the respective measurement and (ii) an initial biological state. For each fixed interval of time points, a first derivative of the normalization relative values in the interval of time points is calculated, thereby forming a plurality of rate transformation values. For each respective set of rate transformation values in the plurality of rate transformation values, a measure of central tendency of the rate transformation values in the set is computed, thereby forming a plurality of average relative transformation values. A maximum metabolic rate and an extent of growth, determined from the normalization relative values and the average relative transformation values, are compared against a lookup table that matches these values to a microorganism type.

Claims

1. A method of identifying a microorganism type in a culture in a vessel, the method comprising:

(A) calculating a normalization relative value for each respective measurement in a plurality of measurements of a biological state of the culture in the vessel, taken at different time points between a first time point and a second time point, between (i) the respective measurement and (ii) an initial biological state of the culture taken at an initial time point, thereby forming a plurality of normalization relative values;

(B) determining, for each respective predetermined fixed interval of time points between the first time point and the second time point, a first derivative of the normalization relative values for measurements of the biological state in the respective predetermined fixed interval of time points, thereby forming a plurality of rate transformation values, wherein the plurality of rate transformation values comprises a plurality of sets of rate transformation values, wherein each respective set of rate transformation values in the plurality of sets of rate transformation values is for a different set of contiguous time points between the first time point and the second time point;

(C) computing, for each respective set of rate transformation values in the plurality of sets of rate transformation values, an average relative transformation value as a measure of central tendency of each of the rate transformation values in the respective set of rate transformation values, thereby computing a plurality of average relative transformation values;

(D) determining a maximum metabolic rate and an extent of growth from the plurality of normalization relative values and the plurality of average relative transformation values; and

(E) determining the microorganism type in the culture in the vessel from the maximum metabolic rate and the extent of growth.

2. The method of claim 1, the method further comprising:

(F) outputting an identification of the microorganism type in the culture in the vessel to a user interface device, a monitor, a computer-readable storage medium, a computer-readable memory, or a local or remote computer system;

or displaying an identification of the microorganism type in the culture.

3. The method of claim 1, wherein the first time point is five or more minutes after the initial time point and the second time point is thirty or more hours after the initial time point.

4. The method of claim 1, wherein the first time point is between 0.5 hours and 3 hours after the initial time point and the second time point is between 4.5 hours and twenty hours after the initial time point.

5. The method of claim 1, wherein the measure of central tendency of the rate transformation values in a first set of rate transformation values in the plurality of sets of rate transformation values comprises:

a geometric mean of each of the rate transformation values in the first set of rate transformation values,

an arithmetic mean of each of the rate transformation values in the first set of rate transformation values,

a median of each of the rate transformation values in the first set of rate transformation values, or

a mode of each of the rate transformation values in the first set of rate transformation values.

6. The method of claim 1, wherein the measurements in the plurality of measurements of the biological state of the culture are each taken of the culture at a periodic time interval between the first time point and the second time point.

7. The method of claim 6, wherein the periodic time interval is an amount of time between one minute and twenty minutes.

8. The method of claim 6, wherein the periodic time interval is an amount of time between five minutes and fifteen minutes.

9. The method of claim 1, wherein the initial biological state of the culture is determined by a fluorescence output of a sensor that is in contact with the culture.

10. The method of claim 9, wherein the amount of fluorescence output of the sensor is affected by CO₂ concentration, O₂ concentration, or pH.

11. The method of claim 1, wherein between 10 and 50,000 measurements of the biological state of the culture in the vessel are in the plurality of measurements of the biological state of the culture.

12. The method of claim 1, wherein between 100 and 10,000 measurements of the biological state of the culture in the vessel are in the plurality of measurements of the biological state of the culture.

13. The method of claim 1, wherein between 150 and 5,000 measurements of the biological state of the culture in the vessel are in the plurality of measurements of the biological state of the culture.

14. The method of claim 1, wherein each respective predetermined fixed interval of time points of step (B) consists of each of the rate transformation values for time points in a time window between the first time point and the second time point, and wherein the time window is a period of time that is between twenty minutes and ten hours.

15. The method of claim 1, wherein each respective predetermined fixed interval of time points of step (B) consists of the rate transformation values for all time points in a time window between the first time point and the second time point in which a biological state of the culture in the vessel was measured, and wherein a duration of the time window is a period of time that is between twenty minutes and two hours.

16. The method of claim 1, wherein each respective predetermined fixed interval of time points of step (B) consists of the rate transformation values for all time points in a time window between the first time point and the second time point in which a biological state of the culture in the vessel was measured, and wherein a duration of the time window is a period of time that is between thirty minutes and ninety minutes.

17. The method of claim 1, wherein each set of rate transformation values in the plurality of rate transformation values consists of between four and twenty contiguous rate transformation values.

18. The method of claim 1, wherein each set of rate transformation values in the plurality of rate transformation values consists of between five and fifteen contiguous rate transformation values.

19. The method of claim 1, wherein there are between five and five hundred average relative transformation values in the plurality of average relative transformation values.

20. The method of claim 1, wherein there are between twenty and one hundred average relative transformation values in the plurality of average relative transformation values.

21. The method of claim 1, wherein a volume of the culture is between 1 ml and 40 ml.

22. The method of claim 1, wherein a volume of the culture is between 2 ml and 10 ml.

23. The method of claim 1, wherein the vessel comprises a sensor composition in fluid communication with the culture, wherein the sensor composition comprises a luminescent compound that exhibits a change in luminescent property, when irradiated with light containing wavelengths that cause said luminescent compound to luminesce, upon exposure to oxygen, wherein the presence of the sensor composition is non-destructive to the culture and wherein the initial biological state of the culture is measured by

irradiating said sensor composition with light containing wavelengths that cause said luminescent compound to luminesce;

and

observing the luminescent light intensity from said luminescent compound while irradiating said sensor composition with said light.

24. The method of claim 23, wherein said luminescent compound is contained within a matrix that is relatively impermeable to water and non-gaseous solutes, but which has a high permeability to oxygen.

25. The method of claim 24, wherein said matrix comprises rubber or plastic.

26. The method of claim 1, wherein the maximum metabolic rate is deemed to be a maximum average relative transformation value in the plurality of average relative transformation values.

27. The method of claim 1, wherein the extent of growth (EG) is determined by the equation:

EG=NR<sub>after—

growth</sub>−

NR<sub>minimum—

growth</sub>

wherein,

NR<sub>after—

growth</sub> is a normalization relative value in the plurality of normalization relative values that was used in the calculation of (i) the first average relative transformation value following a maximum average relative transformation value, (ii) a maximum average relative transformation value, or (iii) the first average relative transformation value preceding a maximum average relative transformation value in the plurality of average relative transformation values; and

NR<sub>minimum—

growth</sub> is a normalization relative value in the plurality of normalization relative values that was used in the calculation of the first average relative transformation value to achieve a threshold value.

28. The method of claim 27, wherein the threshold value is a value between 5 and 100.

29. The method of claim 27, wherein the threshold value is a value between 25 and 75.

30. The method of claim 1, wherein the extent of growth (EG) is determined by the equation:

EG=ARTmax*(time_ARTmax−

time_initial)

wherein,

ARTmax is a maximum average relative transformation value in the plurality of average relative transformation values;

time_ARTmax is a duration of time between (a) the initial time point and (b) a time point when the normalization relative value in the plurality of normalization relative values that was used in the calculation of (i) the first average relative transformation value following the maximum average relative transformation value, (ii) the maximum average relative transformation value, or (iii) the first average relative transformation value preceding the maximum average relative transformation value in the plurality of average relative transformation values was measured; and

time_initial is a duration of time between (i) the initial time point and (ii) a time point when the normalization relative value in the plurality of normalization relative values that was used in the calculation of the first average relative transformation value to achieve a threshold value was measured.

31. The method of claim 30, wherein the threshold value is a value between 5 and 100.

32. The method of claim 30, wherein the threshold value is a value between 25 and 75.

33. The method of claim 1, wherein the extent of growth (EG) is determined by the equation:

EG=[ARTmax*(time_ARTmax−

time_initial)]/time_initial

wherein,

ARTmax is a maximum average relative transformation value in the plurality of average relative transformation values;

time_ARTmax is a duration of time between (a) the initial time point and (b) a time point when the normalization relative value in the plurality of normalization relative values that was used in the calculation of (i) the first average relative transformation value following the maximum average relative transformation value, (ii) the maximum average relative transformation value, or (iii) the first average relative transformation value preceding the maximum average relative transformation value in the plurality of average relative transformation values was measured; and

time_initial is a duration of time between (i) the initial time point and (ii) a time point when the normalization relative value in the plurality of normalization relative values that was used in the calculation of the first average relative transformation value to achieve a threshold value was measured.

34. The method of claim 33, wherein the threshold value is a value between 5 and 100.

35. The method of claim 33, wherein the threshold value is a value between 25 and 75.

36. The method of claim 1, wherein the determining step (E) identifies the microorganism type in the culture in the vessel as (i) a bacterium in the Enterobacteriaceae family or (ii) a bacterium that is not in the Enterobacteriaceae family based upon the maximum metabolic rate and the extent of growth.

37. The method of claim 1, wherein the determining step (E) identifies the microorganism type as bacteria based upon the maximum metabolic rate and the extent of growth.

38. The method of claim 1, wherein the determining step (E) identifies the microorganism type as (i) Enterobacteriaceae, (ii) Staphylococcaceae, (iii) Streptococcus, or (iv) Acinetobacter based upon the maximum metabolic rate and the extent of growth.

39. The method of claim 1, wherein the determining step (E) identifies the microorganism type as a single genera of the Enterobacteriaceae selected from the group consisting of Alishewanella, Alterococcus, Aquamonas, Aranicola, Arsenophonus, Azotivirga, Blochmannia, Brenneria, Buchnera, Budvicia, Buttiauxella, Cedecea, Citrobacter, Dickeya, Edwardsiella, Enterobacter, Erwinia, Escherichia, Ewingella, Griimontella, Hafnia, Klebsiella, Kluyvera, Leclercia, Leminorella, Moellerella, Morganella, Obesumbacterium, Pantoea, Pectobacterium, Candidatus Phlomobacter, Photorhabdus, Plesiomonas, Pragia, Proteus, Providencia, Rahnella, Raoultella, Salmonella, Samsonia, Serratia, Shigella, Sodalis, Tatumella, Trabulsiella, Wigglesworthia, Xenorhabdus, Yersinia, and Yokenella based upon the maximum metabolic rate and the extent of growth.

40. The method of claim 1, wherein the determining step (E) identifies the microorganism type as a single species of Staphylococcaceae selected from the group consisting of Staphylococcus aureus, Staphylococcus caprae, Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus hominis, Staphylococcus lugdunensis, Staphylococcus pettenkoferi, Staphylococcus saprophyticus, Staphylococcus warneri, and Staphylococcus xylosus bacteria based upon the maximum metabolic rate and the extent of growth.

41. The method of claim 1, wherein the determining step (E) determines whether the microorganism is Staphylococcus aureus or coagulase negative staphylococci.

42. The method of claim 1, wherein the determining step (E) identifies the microorganism type as a single species of Streptococcus selected from the group consisting of S. agalactiae, S. bovis, S. mutans, S. pneumoniae, S. pyogenes, S. salivarius, S. sanguinis, S. suis, Streptococcus viridans, and Streptococcus uberis based upon the maximum metabolic rate and the extent of growth.

43. The method of claim 1, wherein the determining step (E) identifies the microorganism type as aerobic based upon the maximum metabolic rate and the extent of growth.

44. The method of claim 1, wherein the determining step (E) identifies the microorganism type as anaerobic based upon the maximum metabolic rate and the extent of growth.

45. The method of claim 1, wherein the initial biological state of the culture is measured by a colorimetric means, a fluorometric means, a nephelometric means, or an infrared means.

46. The method of claim 1, wherein each biological state in the plurality of measurements of the biological state is determined by a colorimetric means, a fluorometric means, a nephelometric means, or an infrared means.

47. The method of claim 1, wherein the culture is a blood culture from a subject.

48. The method of claim 1, wherein the determining step (E) compares the maximum metabolic rate and the extent of growth with a lookup table that matches the maximum metabolic rate and the extent of growth to a microorganism type, thereby determining the microorganism type in the culture in the vessel.

49. An apparatus for identifying a microorganism type in a culture in a vessel, the apparatus comprising a processor and a memory, coupled to the processor, the memory comprising:

a microorganism type determination module comprising;

(A) electronically encoded instructions for calculating a normalization relative value for each respective measurement in a plurality of measurements, taken at a different time points between a first time point and a second time point, between (i) the respective measurement and (ii) an initial biological state of the culture taken at an initial time point, thereby forming a plurality of normalization relative values;

(B) electronically encoded instructions for determining, for each respective predetermined fixed interval of time points between the first time point and the second time point, a first derivative of the normalization relative values for measurements of the biological state in the respective predetermined fixed interval of time points, thereby forming a plurality of rate transformation values, wherein the plurality of rate transformation values comprises a plurality of sets of rate transformation values, wherein each respective set of rate transformation values in the plurality of sets of rate transformation values is for a different set of contiguous time points between the first time point and the second time point;

(C) electronically encoded instructions for computing, for each respective set of rate transformation values in the plurality of sets of rate transformation values, an average relative transformation value as a measure of central tendency of each of the rate transformation values in the respective set of rate transformation values, thereby computing a plurality of average relative transformation values;

(D) electronically encoded instructions for determining a maximum metabolic rate and an extent of growth from the plurality of normalization relative values and the plurality of average relative transformation values; and

(E) electronically encoded instructions for determining the microorganism type in the culture in the vessel from the maximum metabolic rate and the extent of growth.

50. The apparatus of claim 49, the memory further comprising:

a lookup table that comprises matches between (i) a plurality of sets of values, each set of values in the plurality of sets of values comprising a maximum metabolic rate value and an extent of growth, and (ii) a set of microorganism types, wherein, for each set of values in the plurality of sets of values there is a corresponding microorganism type in the set of microorganism types, and wherein

the electronically encoded instructions for determining (E) include instructions for comparing the maximum metabolic rate and the extent of growth with the lookup table that matches the maximum metabolic rate and the extent of growth to a microorganism type, thereby determining the microorganism type in the culture in the vessel.

51. A computer-readable medium storing a computer program product for identifying a microorganism in a culture in a vessel, executable by a computer, wherein the computer program product comprises:

a microorganism type determination module comprising;

(A) electronically encoded instructions for calculating a normalization relative value for each respective measurement in a plurality of measurements, taken at a different time points between a first time point and a second time point, between (i) the respective measurement and (ii) an initial biological state of the culture taken at an initial time point, thereby forming a plurality of normalization relative values;

(B) electronically encoded instructions for determining, for each respective predetermined fixed interval of time points between the first time point and the second time point, a first derivative of the normalization relative values for measurements of the biological state in the respective predetermined fixed interval of time points, thereby forming a plurality of rate transformation values, wherein the plurality of rate transformation values comprises a plurality of sets of rate transformation values, wherein each respective set of rate transformation values in the plurality of sets of rate transformation values is for a different set of contiguous time points between the first time point and the second time point;

(C) electronically encoded instructions for computing, for each respective set of rate transformation values in the plurality of sets of rate transformation values, an average relative transformation value as a measure of central tendency of each of the rate transformation values in the respective set of rate transformation values, thereby computing a plurality of average relative transformation values;

(D) electronically encoded instructions for determining a maximum metabolic rate and an extent of growth from the plurality of normalization relative values and the plurality of average relative transformation values; and

(E) electronically encoded instructions for determining the microorganism type in the culture in the vessel from the maximum metabolic rate and the extent of growth.

52. The computer readable media of claim 51, the computer program product further comprising:

a lookup table that comprises matches between (i) a plurality of sets of values, each set of values in the plurality of sets of values comprising a maximum metabolic rate value and an extent of growth, and (ii) a set of microorganism types, wherein, for each set of values in the plurality of sets of values there is a corresponding microorganism type in the set of microorganism types, and wherein

the electronically encoded instructions for determining (E) include instructions for comparing the maximum metabolic rate and the extent of growth with the lookup table that matches the maximum metabolic rate and the extent of growth to a microorganism type, thereby determining the microorganism type in the culture in the vessel.

53. An apparatus for identifying a microorganism type in a culture in a vessel, the apparatus comprising a processor and a memory, coupled to the processor, the memory comprising:

(A) electronically encoded instructions for obtaining a plurality of sets of values, each set of values in the plurality of sets of values comprising a maximum metabolic rate value and an extent of growth for a different microorganism type in a set of microorganism types; and

(B) electronically encoded instructions for storing a lookup table that comprises matches between (i) the plurality of sets of values and (ii) a set of microorganism types, wherein, for each set of values in the plurality of sets of values there is a corresponding microorganism type in the set of microorganism types.

54. The apparatus of claim 53, the memory further comprising a microorganism type determination module that comprises:

(A) electronically encoded instructions for calculating a normalization relative value for each respective measurement in a plurality of measurements, taken at a different time points between a first time point and a second time point, between (i) the respective measurement and (ii) an initial biological state of the culture taken at an initial time point, thereby forming a plurality of normalization relative values;

(B) electronically encoded instructions for determining, for each respective predetermined fixed interval of time points between the first time point and the second time point, a first derivative of the normalization relative values for measurements of the biological state in the respective predetermined fixed interval of time points, thereby forming a plurality of rate transformation values, wherein the plurality of rate transformation values comprises a plurality of sets of rate transformation values, wherein each respective set of rate transformation values in the plurality of sets of rate transformation values is for a different set of contiguous time points between the first time point and the second time point;

(C) electronically encoded instructions for computing, for each respective set of rate transformation values in the plurality of sets of rate transformation values, an average relative transformation value as a measure of central tendency of each of the rate transformation values in the respective set of rate transformation values, thereby computing a plurality of average relative transformation values;

(D) electronically encoded instructions for determining a maximum metabolic rate and an extent of growth from the plurality of normalization relative values and the plurality of average relative transformation values; and

(E) electronically encoded instructions for comparing the maximum metabolic rate and the extent of growth with the lookup table that matches the maximum metabolic rate and the extent of growth to a microorganism type, thereby determining the microorganism type in the culture in the vessel.

55. A method of identifying a microorganism type in a culture in a vessel, the method comprising:

(A) obtaining a plurality of measurements of the biological state of the culture in the vessel, each measurement in the plurality of measurements taken at a different time point between a first time point and a second time point;

(B) determining, for each respective predetermined fixed interval of time points between the first time point and the second time point, a first derivative of the measurements of the biological state in the respective predetermined fixed interval of time points, thereby forming a plurality of rate transformation values, wherein the plurality of rate transformation values comprises a plurality of sets of rate transformation values, wherein each respective set of rate transformation values in the plurality of sets of rate transformation values is for a different set of contiguous time points between the first time point and the second time point;

(C) computing, for each respective set of rate transformation values in the plurality of sets of rate transformation values, an average relative transformation value as a measure of central tendency of each of the rate transformation values in the respective set of rate transformation values, thereby computing a plurality of average relative transformation values;

(D) determining a maximum metabolic rate and an extent of growth from the plurality of normalization relative values and the plurality of average relative transformation values; and

(E) determining the microorganism type in the culture in the vessel from the maximum metabolic rate and the extent of growth.

56. The method of claim 55, wherein the determining step (E) comprises comparing the maximum metabolic rate and the extent of growth with a lookup table that matches the maximum metabolic rate and the extent of growth to a microorganism type, thereby determining the microorganism type in the culture in the vessel.