METHODS AND KIT FOR ENDOMETRIOSIS SCREENING

• US 20100267003A1
• Filed: 03/29/2007
• Published: 10/21/2010
• Est. Priority Date: 04/03/2006
• Status: Active Grant

• Current Assignee: Diagnostrix AS
• Original Assignee: Oratel Diagnostics, LLC
• Inventors: Goldman, Dorothee
• Granted Patent: US 8,722,349 B2
• Time in Patent Office: Days
• US Class Current: 435/4
• CPC Class Codes: C12Q 1/34 involving hydrolase
• CPC Class Codes: G01N 2333/924 acting on glycosyl compounds (3.2)
• CPC Class Codes: G01N 2800/364 Endometriosis, i.e. non-malignant disorder in which functioning endometrial tissue is present outside the uterine cavity
• CPC Class Codes: G01N 33/84 involving inorganic compounds or pH

Abstract

Methods of screening a bodily sample for endometriosis are provided. The bodily sample is preferably saliva. According to one embodied method, the bodily sample is subjected to a denaturing procedure, and a property of the bodily sample observed after the denaturing procedure is evaluated for the presence or absence of a factor correlating to endometriosis as part of an endometriosis screening procedure. The methods embodied herein are preferably conducted in combinations which permit for the evaluation of at least two different physiological factors correlating to endometriosis, because women with endometriosis do not always share the same factors. Also provided are screening kits, assay methods, and systems for endometriosis screening.

Claims

1. A method of screening for endometriosis in a female subject, comprising:

performing a first assay on a first non-invasive bodily sample to evaluate for a first physiological factor correlating to endometriosis;

performing a second assay on a second non-invasive bodily sample to evaluate for a second physiological factor correlating to endometriosis, the first and second physiological factors differing from one another; and

evaluating the assays to determine whether the female subject has endometriosis based on the presence or absence of the physiological factors.

2. The method of claim 1, wherein the bodily samples comprise saliva.

3. The method of claim 1, wherein the first and second physiological factors are selected from immune factors and inflammatory factors.

4. The method of claim 1, wherein the first assay comprises:

subjecting a bodily sample to a denaturing procedure; and

evaluating a property of the bodily sample after the denaturing procedure as part of an endometriosis screening procedure.

5. The method of claim 4, wherein said subjecting of the bodily sample to the denaturing procedure comprises subjecting the bodily sample to at least one freezing and thawing cycle.

6. The method of claim 4, further comprising:

measuring a property of the bodily sample prior and subsequent to the denaturing procedure;

comparing the property measured prior and subsequent to the denaturing procedure as part of the endometriosis screening procedure; and

evaluating for endometriosis in the female subject.

7. The method of claim 4, wherein said subjecting of the bodily sample to the denaturing procedure comprises subjecting the bodily sample to at least one freezing and thawing cycle, and wherein said measuring comprising obtaining a first pH measurement of the bodily sample prior to the freezing and thawing cycle and obtaining a second pH measurement of the bodily sample subsequent to the freezing and thawing cycle.

8. The method of claim 7, wherein said comparing comprises subtracting the second pH measurement from the first pH measurement to obtain a difference value, and wherein said evaluating comprises interpreting a pH difference value of less than 0.2 as an indicator that the female subject has endometriosis and interpreting a pH difference value of greater than 0.2 as an indicator that the female subject does not have endometriosis.

9. The method of claim 4, further comprising:

collecting the bodily sample and additional bodily samples periodically over at least five consecutive days;

subjecting the bodily samples to freezing and thawing cycles;

measuring first pH measurements of the bodily samples prior to the freezing and thawing cycle and obtaining second pH measurements of the bodily samples subsequent to the freezing and thawing cycle;

for each of the bodily samples, subtracting the second pH measurements from the first pH measurements to obtain difference values; and

interpreting an occurrence of pH difference values of less than 0.2 for bodily samples collected over five or more consecutive days as an indicator that the female subject has endometriosis, and interpreting a nonoccurrence of more than four pH changes of less than 0.2 for bodily fluid samples collected over five consecutive days as an indicator that the female subject does not have endometriosis.

10. The method of claim 6, wherein said measuring comprises obtaining a first set of optical absorbency measurements of the bodily sample prior to the denaturing procedure and obtaining a second set of optical absorbency measurements of the bodily sample subsequent to the denaturing procedure.

11. The method of claim 4, wherein:

the denaturing procedure comprises dehydrating the bodily sample; and

the property comprises crystalline formations of the dehydrated bodily sample.

12. The method of claim 11, wherein said evaluating comprises:

obtaining digital micro-photographs of a plurality of dehydrated bodily samples taken over all menstrual cycle phases, the digital microphotographs capturing the crystalline formations of the dehydrated bodily sample; and

comparing the crystalline formations captured in the digital microphotographs to the reference microphotographs.

13. The method of claim 4, further comprising:

combining the bodily sample with a flavonoid pigment;

measuring first and second optical density values of the combination of the bodily sample and the flavonoid pigment at first and second wavelengths, respectively, prior to the denaturing procedure;

measuring third and fourth optical density values of the combination of the bodily sample and the flavonoid pigment at first and second wavelengths, respectively, subsequent to the denaturing procedure; and

comparing the optical density values prior and subsequent to the denaturing procedure as part of the endometriosis screening procedure.

14. The method of claim 1, wherein the first assay comprises:

providing a mixture comprising a bodily sample and a flavonoid pigment;

measuring first and second optical density values of the mixture at first and second wavelengths, respectively;

calculating a mathematical relationship value between the optical density values; and

comparing the mathematical relationship value to a reference scale as part of an endometriosis screening procedure.

15. The method of claim 14, wherein the mathematical relationship value comprises a ratio of the first and second optical densities.

16. The method of claim 14, wherein the first and second wavelengths are in a range of 400 nm to 700 nm.

17. The method of claim 16, wherein the first and second wavelengths are about 560 nm and about 538 nm, respectively.

18. The method of claim 14, wherein:

the mathematical relationship value comprises a ratio of the first optical density peak value to the second optical density peak value; and

said comparing comprises interpreting a ratio value in taken in the luteal phase of menstrual cycle of 1 to 1.2 as an indicator that the female subject has endometriosis, and interpreting a ratio value taken in the luteal phase of menstrual cycle of greater than 1.3 as an indicator that the female subject does not have endometriosis.

19. The method of claim 17, further comprising:

collecting bodily samples over a plurality of successive days in the luteal phase of a menstrual cycle;

subjecting each of the bodily samples to said measuring, calculating and comparing steps;

interpreting a ratio value of 1.0 to 1.10 occurring at a frequency of greater than 60% as an indicator that the female subject has endometriosis; and

interpreting a ratio value of 1.10 to 1.2 occurring at a frequency of less than 15% as an indicator that the female subject does not have endometriosis.

20. The method of claim 3, wherein the first assay comprises:

combining a saliva sample with a quercetin pigment;

treating the sample and quercetin pigment combination with diluted tincture of iodine and detecting for a color response; and

evaluating the color response as part of an endometriosis screening procedure.

21. The method of claim 20, further comprising embedding the pigment in a solid matrix.

22. The method of claim 20, wherein the solid matrix comprises cellulose.

23. The method of claim 1, wherein the first assay comprises:

filtering a bodily fluid sample;

subjecting the bodily fluid sample to a testing procedure prior and subsequent to said filtering and observing a first test result and a second test result, respectively; and

evaluating a difference, if any, in the first and second test results as part of an endometriosis screening procedure.

24. The method of claim 23, further comprising combining the bodily fluid sample with a color response system and observing color as the test results, wherein said evaluating comprises comparing the first test result and the second test result to determine whether filtering caused a color change to the treated bodily fluid sample.

25. The method of claim 1, wherein the first assay comprises:

combining a bodily fluid sample with quercetin;

subjecting the bodily fluid sample to a chromatography procedure; and

evaluating results of the chromatography procedure as part of an endometriosis screening procedure.

65. A kit, comprising:

a platform equipped for receiving bodily samples and carrying out a method of claim 1 to evaluate for a physiological factor, the presence or absence of which correlates to endometriosis.

26. A method of screening for endometriosis in a female subject, comprising:

subjecting a bodily sample to a denaturing procedure; and

evaluating a property of the bodily sample after the denaturing procedure as part of an endometriosis screening procedure.

27. The method of claim 26, wherein said subjecting of the bodily sample to the denaturing procedure comprises subjecting the bodily sample to at least one freezing and thawing cycle.

28. The method of claim 26, further comprising:

measuring a property of the bodily sample prior and subsequent to the denaturing procedure;

comparing the property measured prior and subsequent to the denaturing procedure as part of the endometriosis screening procedure; and

evaluating for endometriosis in the female subject.

29. The method of claim 28, wherein said subjecting of the bodily sample to the denaturing procedure comprises subjecting the bodily sample to at least one freezing and thawing cycle, and wherein said measuring comprising obtaining a first pH measurement of the bodily sample prior to the freezing and thawing cycle and obtaining a second pH measurement of the bodily sample subsequent to the freezing and thawing cycle.

30. The method of claim 29, wherein said comparing comprises subtracting the second pH measurement from the first pH measurement to obtain a difference value, and wherein said evaluating comprises interpreting a pH difference value of less than 0.2 as an indicator that the female subject has endometriosis and interpreting a pH difference value of greater than 0.2 as an indicator that the female subject does not have endometriosis.

31. The method of claim 30, wherein the bodily sample comprises saliva.

32. The method of claim 28, further comprising:

collecting the bodily sample and additional bodily samples periodically over at least five consecutive days;

subjecting the bodily samples to freezing and thawing cycles;

measuring first pH measurements of the bodily samples prior to the freezing and thawing cycle and obtaining second pH measurements of the bodily samples subsequent to the freezing and thawing cycle;

for each of the bodily samples, subtracting the second pH measurements from the first pH measurements to obtain difference values; and

interpreting an occurrence of pH difference values of less than 0.2 for bodily samples collected over five or more consecutive days as an indicator that the female subject has endometriosis, and interpreting a nonoccurrence of more than four pH changes of less than 0.2 for bodily fluid samples collected over five consecutive days as an indicator that the female subject does not have endometriosis.

33. The method of claim 28, wherein said measuring comprises obtaining a first set of optical absorbency measurements of the bodily sample prior to the denaturing procedure and obtaining a second set of optical absorbency measurements of the bodily sample subsequent to the denaturing procedure.

34. The method of claim 26, wherein:

the denaturing procedure comprises dehydrating the bodily sample; and

the property comprises crystalline formations of the dehydrated bodily sample.

35. The method of claim 34, wherein said evaluating comprises:

obtaining digital micro-photographs of a plurality of dehydrated bodily samples taken over all menstrual cycle phases, the digital microphotographs capturing the crystalline formations of the-dehydrated bodily sample; and

comparing the crystalline formations captured in the digital microphotographs to the reference microphotographs.

36. The method of claim 34, wherein the bodily sample comprises saliva.

37. The method of claim 26, further comprising:

combining the bodily sample with a flavonoid pigment;

measuring first and second optical density values of the combination of the bodily sample and the flavonoid pigment at first and second wavelengths, respectively, prior to the denaturing procedure;

measuring first and second optical density values of the combination of the bodily sample and the flavonoid pigment at first and second wavelengths, respectively, subsequent to the denaturing procedure; and

comparing the optical density values prior and subsequent to the denaturing procedure as part of the endometriosis screening procedure.

38. The method of claim 37, wherein the bodily sample comprises saliva.

39. A method of screening for endometriosis in a female subject, comprising:

providing a mixture comprising a bodily sample and a flavonoid pigment;

measuring first and second optical density values of the mixture at first and second wavelengths, respectively;

calculating a mathematical relationship value between the optical density values; and

comparing the mathematical relationship value to a reference scale as part of an endometriosis screening procedure.

40. The method of claim 39, wherein the bodily sample comprises saliva.

41. The method of claim 40, wherein the mathematical relationship value comprises a ratio of the first and second optical densities.

42. The method of claim 39, wherein the first and second wavelengths are in a range of 400 nm to 700 nm.

43. The method of claim 42, wherein the first and second wavelengths are about 560 nm and about 538 nm, respectively.

44. The method of claim 43, wherein the bodily sample comprises saliva.

45. The method of claim 44, wherein:

the mathematical relationship value comprises a ratio of the first optical density peak value to the second optical density peak value; and

said comparing comprises interpreting a ratio value in taken in the luteal phase of menstrual cycle of 1 to 1.2 as an indicator that the female subject has endometriosis, and interpreting a ratio value taken in the luteal phase of menstrual cycle of greater than 1.3 as an indicator that the female subject does not have endometriosis.

46. The method of claim 44, further comprising:

collecting bodily samples over a plurality of successive days in the luteal phase of a menstrual cycle;

subjecting each of the bodily samples to said measuring, calculating and comparing steps;

interpreting a ratio value of 1.0 to 1.10 occurring at a frequency of greater than 60% as an indicator that the female subject has endometriosis; and

interpreting a ratio value of 1.10 to 1.2 occurring at a frequency of less than 15% as an indicator that the female subject does not have endometriosis.

47. A method of screening for endometriosis in a female subject, comprising:

combining a saliva sample with a quercetin pigment;

treating the sample and the quercetin pigment with diluted tincture of iodine and detecting for a color response; and

evaluating the color response as part of an endometriosis screening procedure.

48. The method of claim 47, further comprising collecting the bodily fluid samples periodically over multiple days, and subjecting the samples to said treating and evaluating steps.

49. The method of claim 47, further comprising embedding the pigment in a solid matrix.

50. The method of claim 47, wherein the solid matrix comprises cellulose.

51. A method of screening for endometriosis in a female subject, comprising:

filtering a bodily fluid sample;

subjecting the bodily fluid sample to a testing procedure prior and subsequent to said filtering and observing a first test result and a second test result, respectively; and

evaluating a difference, if any, in the first and second test results as part of an endometriosis screening procedure.

52. The method of claim 51, further comprising combining the bodily fluid sample with a color response system and observing color as the test results, wherein said comparing comprises comparing the first test result and the second test result to determine whether filtering caused a color change to the treated bodily fluid sample.

53. The method of claim 51, wherein the bodily fluid sample comprises saliva.

54. A method of screening for endometriosis in a female subject, comprising:

combining a bodily fluid sample with quercetin;

subjecting the bodily fluid sample to a chromatography procedure; and

evaluating results of the chromatography procedure as part of an endometriosis screening procedure.

55. An assay method comprising:

performing a first assay on a first non-invasive bodily sample;

evaluating the first assay for a first physiological factor, the presence or absence of which in the first non-invasive bodily sample correlates to endometriosis;

performing a second assay on a second non-invasive bodily sample; and

evaluating the second assay for a second physiological factor, the presence or absence of which in the second non-invasive bodily sample correlates to endometriosis, the first and second physiological factors differing from one another.

56. An assay method, comprising:

performing a denaturing assay on a non-invasive bodily sample; and

evaluating a property of the bodily sample after the denaturing assay.

57. The assay method of claim 56, wherein the denaturing assay comprises at least one freezing and thawing cycle.

58. The assay method of claim 57, further comprising obtaining a first pH measurement of the non-invasive bodily sample prior to the freezing and thawing cycle and obtaining a second pH measurement of the non-invasive bodily sample subsequent to the freezing and thawing cycle.

59. The assay method of claim 56, wherein:

the denaturing assay comprises dehydrating the bodily sample; and

the property comprises crystalline formations of the dehydrated bodily sample.

60. The assay method of claim 56, further comprising:

combining the bodily sample with a flavonoid pigment;

measuring first and second optical density values of the combination of the bodily sample and the flavonoid pigment at first and second wavelengths, respectively, prior to the denaturing assay;

measuring first and second optical density values of the combination of the bodily sample and the flavonoid pigment at first and second wavelengths, respectively, subsequent to the denaturing assay; and

comparing the optical density values prior and subsequent to the denaturing assay.

61. An assay method, comprising:

measuring first and second optical density values of a mixture comprising a bodily sample and a flavonoid pigment at first and second wavelengths, respectively;

calculating a mathematical relationship value between the optical density values.

62. An assay method, comprising:

combining a non-invasive bodily sample with a quercetin pigment;

treating the sample and the quercetin pigment with diluted tincture of iodine and detecting for a color response; and

evaluating the color response.

63. An assay method, comprising:

combining a non-invasive bodily fluid sample with quercetin;

subjecting the bodily fluid sample to a chromatography procedure; and

evaluating results of the chromatography procedure.

64. A kit, comprising:

a platform for receiving bodily samples and carrying out a first assay on a first non-invasive bodily sample to evaluate for a first physiological factor correlating to endometriosis and a second assay on a second non-invasive bodily sample to evaluate for a second physiological factor correlating to endometriosis, the first and second physiological factors differing from one another.