Spectrophotometric system and method for the identification and characterization of a particle in a bodily fluid

• US 7,027,134 B1
• Filed: 09/03/2004
• Issued: 04/11/2006
• Est. Priority Date: 02/08/1995
• Status: Expired due to Term

• Current Assignee: University of South Florida (State University System of Florida)
• Original Assignee: University of South Florida (State University System of Florida)
• Inventors: Garcia-Rubio, Luis H., Cutolo, Edward P., Peguero, Alfredo, Alupoaei, Catalina E., Harris, Willard, Leparc, German Felix
• Primary Examiner(s): Stafira, Michael P.
• Assistant Examiner(s): VALENTIN, JUAN D
• Application Number:
  US10/934,601
• Time in Patent Office: 585 Days
• Field of Search: 356 39- 41, 356/335, 356/337, 356/441, 356/433, 356300-334, 600/310, 600/314
• US Class Current: 356/39
• CPC Class Codes: G01N 21/31 Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry G01N21/72 takes precedence
• CPC Class Codes: G01N 21/49 within a body or fluid
• CPC Class Codes: G01N 33/6803 General methods of protein analysis not limited to specific proteins or families of proteins
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The present invention provides a method and apparatus for the detection of an infectious disease or disorder in a fluid, such as a mammalian blood sample, the detection of a specific protein in a urine sample, or the detection of a particle in a plasma. The identification of the particles of interest is enable by taking a transmission spectrum of a test sample in at least a portion of the ultraviolet, visible, near-infrared portion of the spectrum and comparing the spectrum with a standard sample spectrum. From the comparison it is then determined whether the fluid from the test sample contains an particle of interest, and an identity of the particle of interest is determined. Spectroscopic and multiwavelength turbidimetry techniques provide a rapid, inexpensive, and convenient means for diagnosis. The comparison and determination steps may be performed visually or by spectral deconvolution.

Claims

1. A method for detecting the presence of a protein in a test urine sample, the method comprising the steps of:

aggregating a test urine sample;

taking a transmission spectrum of the test urine sample in at least a part of the ultraviolet visible—

near-infrared range of the electromagnetic spectrum;

comparing the transmission spectrum with a theoretically calculated standard urine sample transmission spectrum known to be free from the protein; and

determining from the comparison whether the urine from the test sample contains the protein.

2. The method recited in claim 1, wherein the protein comprises an agent that alters at least one of a shape, a size, and a chemical composition of a normal urine component.

3. The method recited in claim 1, wherein the comparing step comprises identifying a difference in at least one of a peak height, a peak presence, and a slope between the standard sample and the test sample.

4. The method recited in claim 1, wherein the step of aggregating the test urine sample results in modifications in the absorption and scattering properties of the sample.

5. The method recited in claim 1, wherein the step of aggregating the test urine sample further comprises the addition of a salt to the test urine sample.

6. The method of claim 5, wherein the salt is trichloroacetic acid.

7. The method recited in claim 1, wherein the transmission spectrum has a resolution of at least 2 nm.

8. The method recited in claim 1, wherein the comparing step comprises identifying a feature of the standard spectrum known to change in a presence of the protein and the determining step comprises analyzing the test spectrum for a change in the identified feature.

9. The method recited in claim 1, further comprising the step, prior to the comparing step, of normalizing the standard spectrum and the test spectrum for facilitating the comparing step.

10. The method recited in claim 1, further comprising the step, of adjusting a path length in the spectrum taking step to an optical density range in which the response of the spectrometer is substantially linearly related to the concentration of the sample.

11. The method of claim 1, wherein the protein is albumin.

12. The method of claim 1, wherein the protein is creatinine.

13. A method of quantifying a protein in a test urine sample comprising the steps of:

aggregating a test urine sample;

taking a transmission spectrum of the test urine sample in at least a portion of the ultraviolet visible near-infrared range of the electromagnetic spectrum;

deconvolving the spectrum into absorption and scattering components; and

determining from the deconvolution a presence and a concentration of a protein in the urine sample.

14. The method recited in claim 13, wherein the protein is albumin.

15. The method recited in claim 13, wherein the protein is creatinine.

16. The method recited in claim 13, wherein the step of aggregating the test urine sample results in modifications in the absorption and scattering properties of the sample.

17. The method recited in claim 13, wherein the step of aggregating the test urine sample further comprises the addition of a salt to the test urine sample.

18. The method of claim 17, wherein the salt is trichloroacetic acid.

19. The method recited in claim 13, wherein the spectrum taking step comprises taking a spectrum in a range of approximately 220–

900 nm.

20. The method recited in claim 13, wherein the deconvolving step comprises utilizing a calibration approach based on correlation.

21. The method recited in claim 13, wherein the deconvolving step comprises utilizing absorption and scattering theories.

22. A urine test kit for detecting the presence of a protein, the test kit comprising:

aggregating reagents;

a spectrophotometer for taking a transmission spectrum of a test urine sample in at least a portion of the ultraviolet visible near-infrared range of the electromagnetic spectrum;

an analyzer for accessing a standard spectrum from a urine sample known to be free from the protein for comparing the test urine sample transmission spectrum with the standard urin sample spectrum to determine whether the urine from the test sample contains the protein.

23. The urine test kit recited in claim 22, wherein the analyzer comprises a processor and a storage medium in electronic communication with the processor, the storage medium having stored thereon a database of standard spectra.

24. The urine test kit recited in claim 22, wherein the analyzer further comprises a software package resident on the processor having a routine for performing spectral deconvolution of the standard spectrum and the test spectrum, for identifying features of the test spectrum associated with the protein.

25. The urine test kit recited in claim 22, wherein the analyzer further comprises an output device in electronic communication with the processor for providing the standard spectrum and the test spectrum in visible form.

26. The urine test kit recited in claim 22, wherein the aggregating reagent is a salt.

27. The urine test kit recited in claim 22, wherein the aggregating reagent is trichloroacetc acid.

28. The urine test kit recited in claim 22, wherein the protein is albumin.

29. The urine test kit recited in claim 22, wherein the protein is creatinine.

30. A method for detecting the presence of a particle in a plasma test sample, the method comprising the steps of:

taking a transmission spectrum of the test plasma sample in at least a part of the ultraviolet visible—

near-infrared range of the electromagnetic spectrum;

comparing the transmission spectrum of the test plasma sample with a theoretically calculated standard plasma sample transmission spectrum known to be free from the particle of interest; and

determining from the comparison whether the plasma from the test sample contains the particle of interest.

31. The method recited in claim 30, wherein the particle comprises an agent that alters at least one of a shape, a size, and a chemical composition of a normal plasma component.

32. The method recited in claim 30, wherein the comparing step comprises identifying a difference in at least one of a peak height, a peak presence, and a slope between the standard sample and the test sample.

33. The method recited in claim 30, wherein the particle is a platelet.

34. The method recited in claim 30, wherein the particle is a microorganism.

35. The method recited in claim 30, wherein the transmission spectrum has a resolution of at least 2 nm.

36. The method recited in claim 30, wherein the comparing step comprises identifying a feature of the standard spectrum known to change in a presence of the particle and the determining step comprises analyzing the test spectrum for a change in the identified feature.

37. The method recited in claim 30, further comprising the step, prior to the comparing step, of normalizing the standard spectrum and the test spectrum for facilitating the comparing step.

38. The method recited in claim 30, further comprising the step, of adjusting a path length in the spectrum taking step to an optical density range in which the response of the spectrometer is substantially linearly related to the concentration of the sample.

39. A method of quantifying a particle in a test plasma sample comprising the steps of:

taking a transmission spectrum of the test plasma sample in at least a portion of the ultraviolet visible near-infrared range of the electromagnetic spectrum;

deconvolving the spectrum into absorption and scattering components; and

determining from the deconvolution a presence and a concentration of a particle in the plasma sample.

40. The method recited in claim 39, wherein the particle is a platelet.

41. The method recited in claim 39, wherein the particle is a microorganism.

42. The method recited in claim 39, wherein the spectrum taking step comprises taking a spectrum in a range of approximately 220–

900 nm.

43. The method recited in claim 39, wherein the deconvolving step comprises utilizing a calibration approach based on correlation.

44. The method recited in claim 39, wherein the deconvolving step comprises utilizing absorption and scattering theories.

45. A plasma test kit for detecting the presence of a particle, the test kit comprising:

a spectrophotometer for taking a transmission spectrum of a test plasma sample in at least a portion of the ultraviolet visible near-infrared range of the electromagnetic spectrum;

an analyzer for accessing a standard spectrum from a plasma sample known to be free from the particle for comparing the test plasma sample transmission spectrum with the standard plasma sample spectrum to determine whether the plasma from the test sample contains the particle of interest.

46. The plasma test kit recited in claim 45, wherein the analyzer comprises a processor and a storage medium in electronic communication with the processor, the storage medium having stored thereon a database of standard spectra.

47. The plasma test kit recited in claim 45, wherein the analyzer further comprises a software package resident on the processor having a routine for performing spectral deconvolution of the standard spectrum and the test spectrum, for identifying features of the test spectrum associated with the particle.

48. The plasma test kit recited in claim 45, wherein the analyzer further comprises an output device in electronic communication with the processor for providing the standard spectrum and the test spectrum in visible form.

49. The plasma test kit recited in claim 45, wherein the particle of interest is a platelet.

50. The urine test kit recited in claim 45, wherein the particle of interest is a microorganism.