Method and system for optically performing an assay to determine a medical condition

• US 20050021235A1
• Filed: 05/30/2002
• Published: 01/27/2005
• Est. Priority Date: 05/30/2001
• Status: Abandoned Application

• Current Assignee: Ischemia Technologies, Inc. (Abbott Laboratories Incorporated)
• Original Assignee: Ischemia Technologies, Inc. (Abbott Laboratories Incorporated)
• Inventors: Curtis, C. Gerald, Bar-Or, David, Bar-Or, Raphael
• Application Number:
  US10/477,384
• Publication Number:
  US 20050021235A1
• Time in Patent Office: Days
• US Class Current: 702/19
• CPC Class Codes: A61B 5/0071 by measuring fluorescence emission
• CPC Class Codes: A61B 5/0075 by spectroscopy, i.e. measuring spectra, e.g. Raman spectroscopy, infrared absorption spectroscopy A61B5/0071 takes precedence
• CPC Class Codes: A61B 5/0084 for introduction into the body, e.g. by catheters A61B5/1459 takes precedence
• CPC Class Codes: G01N 2021/6423 Spectral mapping, video display
• 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 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 2201/129 Using chemometrical methods
• CPC Class Codes: G01N 2333/76 Assays involving albumins other than in routine use for blocking surfaces or for anchoring haptens during immunisation
• CPC Class Codes: G01N 2800/2871 Cerebrovascular disorders, e.g. stroke, cerebral infarct, cerebral haemorrhage, transient ischemic event
• CPC Class Codes: G01N 2800/32 Cardiovascular disorders
• CPC Class Codes: G01N 33/683 involving metal ions
• CPC Class Codes: G01N 33/84 involving inorganic compounds or pH

Abstract

A method and system are disclosed for detecting a medical condition wherein a blood or plasma sample is combined with a metal such as cobalt and optically analyzed for an optical distinction that identifies the medical condition. The invention is useful for diagnosing medical conditions such as ischemia. Moreover, the diagnoses of patient samples according to the invention may be enhanced by developing a mathematical model based on signal processing techniques such as principal component analysis on the data obtained in patient studies.

Claims

1. A method for detecting a medical condition comprising:

providing a patient fluid sample divided into first and second portions, and combining a substance for providing free metal ions with the first portion of the sample;

irradiating both the first and second portions of the sample with light;

determining absorbance values for the first and second portions;

obtaining a differential absorbance value from the first and second portions;

analyzing the differential absorbance value for determining one or more characteristics that are indicative of whether the medical condition is present;

wherein said analyzing step uses principal component analysis for reducing a dimension of the differential absorbance value.

2. The method of claim 1, wherein the medical condition is ischemia.

3. The method of claim 1, wherein the metal ion is cobalt ion.

4. A method of diagnosing an ischemic event comprising:

a) providing a first and second patient sample comprising albumin;

b) adding to the first patient sample a metal ion, whereby the metal ion binds to the albumin;

c) conducting optical analyses of the first and second patient samples to generate signals or spectra, respectively;

d) measuring the amount of metal bound to the albumin by comparing the signals or spectra of step (c) to generate a differential signal or spectra; and

e) comparing the differential signal or spectra to a standard curve or mathematical model that correlates the differential signal or spectra to amount of metal bound to albumin, whereby an ischemic event may be diagnosed if the measured amount of metal bound to albumin is below a defined value.

5. The method of claim 4, wherein the patient samples are serum.

6. The method of claim 4, wherein the first and second patient samples are provided by dividing an original patient sample.

8. The method of claim 4, wherein the metal ion binds to the N-terminus of the albumin.

9. The method of claim 4, wherein the optical analyses are absorbance spectroscopy and the analyses is conducted in the range of 300-450 nm.

10. The method of claim 4, wherein the optical analyses comprise fluorescence spectroscopy, said method further comprising:

adding to the first patient sample a fluorescent dye in step (b), wherein the dye binds to said metal ion and wherein the fluorescence signal changes as a function of whether the metal ion is unbound or bound to the albumin.

11. The method of claim 4, further comprising:

analyzing the differential signal or spectra of step (d) using principal component analysis.

7. The method of claim 43, wherein the metal ion is cobalt ion.

12. A method for diagnosing an ischemic event, comprising:

a) adding a metal ion and a fluorescent dye to a patient sample comprising albumin, whereby the dye binds to the metal ion which may bind to the albumin;

b) measuring the metal bound to the albumin by measuring a fluorescent signal of the sample, wherein the fluorescent signal changes as a function of whether the metal ion is unbound or bound to the albumin; and

c) comparing the fluorescent signal to a standard curve or mathematical model that correlates the fluorescent signal to an amount of metal ion bound to albumin, whereby the measurement of metal ion bound to albumin below a defined value may be diagnostic for an ischemic event.

13. The method of claim 12 wherein the metal ion and fluorescent dye are added as a conjugate.

14. The method of claim 12, wherein the fluorescent signal is quenched or shifts to a different wavelength when the dye is bound to a metal ion that is bound to the albumin.

15. The method of claim 12, wherein the metal ion is a cobalt ion.

16. The method of claim 12, wherein the fluorescent dye is selected from the group consisting of Cumarin, Rhodamine and Newport green.

17. A method of rapidly diagnosing an ischemic event comprising:

a) providing a first and second patient sample comprising albumin;

b) adding to the first patient sample a metal ion, whereby the metal ion binds to the albumin in a reaction that reaches equilibrium at a predetermined time;

c) conducting, during a defined time interval prior to achievement of equilibrium, optical analyses of the first and second patient samples to generate first and second signals or spectra, respectively, for each sample at selected time points during the defined time interval;

d) measuring the rate of change of amount of metal bound to the albumin over the defined time interval by comparing the first and second signals or spectra for each time point to generate differential signals or spectra for each time point in the time interval;

e) calculating a rate of change in the differential signals or spectra over the time interval;

f) and comparing the rate of change of signal or spectra to a standard curve or mathematical model that correlates rate of change with projected metal bound to albumin at equilibrium, whereby an ischemic event may be diagnosed if the projected amount of metal bound to albumin is below a defined value.

18. A method of rapidly diagnosing an ischemic event, comprising:

a) adding a metal ion and a fluorescent dye to a patient sample comprising albumin, whereby the dye binds to the metal ion which binds to the albumin in a reaction that reaches equilibrium at a predetermined time, wherein the fluorescent dye'"'"'s signal changes as a function of whether the metal ion is unbound or bound to the albumin;

b) measuring the rate of change of metal bound to the albumin by measuring the fluorescent signal of the sample at selected time points over a time interval that is prior to achievement of equilibrium;

c) calculating the rate of change of the fluorescent signal over the time interval; and

d) comparing the rate of change of the fluorescent signal to a standard curve or mathematical model that correlates the rate of change in fluorescent signal to a projected amount of metal ion bound to albumin at equilibrium, whereby ischemia may be diagnosed if the measured rate of change of metal ion bound to albumin is below a defined value.

19. A method for diagnosing an ischemic event comprising:

(a) providing a patient sample comprising albumin, a portion of which may be N-terminally modified;

(b) measuring the N-terminally modified albumin by measuring absorbance of the sample, and comparing the absorbance to a standard curve or mathematical model that correlates the absorbance to a ratio of modified to unmodified albumin, wherein an ischemic event may be diagnosed if the ratio is below a defined value.

20. The method of claim 19, wherein the patient sample comprises whole blood, serum or plasma provided in a sample container and the absorbance is measured with a spectral probe placed in the sample.

21. The method of claim 19, wherein the patient sample comprises whole blood in the patient'"'"'s blood vessel and the absorbance is measured with a spectral probe placed in the blood vessel.

22. An instrument for detecting a medical condition, comprising:

a spectral probe having a tip for insertion into a patient fluid sample and receiving spectral light from the patient fluid sample;

a spectrophotometer coupled to said spectral probe for quantifying each frequency of spectral light received by said spectral probe and outputting a signal representative of the quantity of each frequency of spectral light;

a computing unit, comprising;

an input coupled to receive the signal from said spectrophotometer;

a memory for storing a model representing spectral light data obtained from a first set of patients known to have the medical condition and a second set of individuals known to not have the medical condition, whereby the model includes a value identified with a high probability of the presence of the medical condition;

a processor programmed to execute instructions for;

comparing the quantity of each frequency of spectral light from the patient with corresponding data in the stored model; and

determining whether the quantity of each frequency of spectral light is indicative of the presence of the medical condition in the patient; and

an output to provide the determination to a user.

23. A method for providing an instrument for diagnosing a medical condition in a patient, comprising:

obtaining a control fluid sample from a first plurality of control individuals known to have the medical condition;

obtaining a control fluid sample from a second plurality of control individuals known to not have the medical condition;

dividing each control fluid sample into first and second portions;

combining a substance for providing free metal ions with the first portion of each control fluid sample;

irradiating both the first and second portions of each control fluid sample with light;

determining absorbance values for the first and second portions of each control fluid sample;

obtaining a differential absorbance value from the first and second portions of each control fluid sample;

generating a principal component analysis model of the obtained differential absorbance values, the principal component analysis model including a value indicative of the presence of the medical condition;

storing the generated principal component analysis model in a computer readable format;

providing computer executable instructions for;

providing a differential absorbance value, determined from first and second portions of a patient fluid sample obtained from a patient, said first portion having been combined with free metal ions, and comparing said differential value with the stored principal component analysis model;

in response to the comparing step, determining whether the differential absorbance value of the patient fluid sample is indicative of the presence of the medical condition.