Methods, Devices, and Systems for Glycated Hemoglobin Analysis

• US 20110269147A1
• Filed: 07/20/2009
• Published: 11/03/2011
• Est. Priority Date: 07/18/2008
• Status: Abandoned Application

• Current Assignee: Polymer Technology Systems Incorporated (Sinocare, Inc.)
• Original Assignee: Bayer Healthcare LLC (Bayer AG)
• Inventors: Chinnayelka, Swetha
• Application Number:
  US13/054,543
• Publication Number:
  US 20110269147A1
• Time in Patent Office: Days
• US Class Current: 435/7.1
• 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 2333/805 Haemoglobins; Myoglobins
• CPC Class Codes: G01N 2800/22 Haematology
• CPC Class Codes: G01N 33/542 with steric inhibition or signal modification, e.g. fluorescent quenching
• CPC Class Codes: G01N 33/721 Haemoglobin
• CPC Class Codes: G01N 33/723 Glycosylated haemoglobin

Abstract

Methods, devices, and systems for measuring the concentration of glycated hemoglobin (HbA1c) in a blood sample are provided. A blood sample may be contacted with a lysing agent, an assay to measure total Hb, and an assay to measure HbA1c, which comprises donor and acceptor dyes attached to a Hb specific antibody and a HbA1c analog. Preferably, the emission spectrum of the donor dye overlaps with the excitation spectrum of the acceptor dye. The concentration of HbA1c in the sample may be correlated with an increase in donor dye and/or a decrease in acceptor dye fluorescence in the presence of HbA1c as compared to the donor and/or the acceptor dye fluorescence in the absence of HbA1c.

Claims

1. A method for determining the concentration of glycated hemoglobin (HbA1c) in a sample, comprising:

contacting the sample with a red blood cell lysing mixture;

contacting the sample with a total hemoglobin assay;

contacting the sample with a HbA1c assay including one of

a fluorescent donor dye conjugated to a HbA1c analog and a fluorescent acceptor dye attached to a Hb-specific antibody, and

a fluorescent donor dye attached to a Hb-specific antibody and a fluorescent acceptor dye attached to a HbA1c analog,

where the emission spectrum of the fluorescent donor dye overlaps with the excitation spectrum of the fluorescent acceptor dye; and

correlating the concentration of HbA1c in the sample with a change in the intensity of at least one of donor dye fluorescence and acceptor dye fluorescence.

2. The method of claim 1, where the total hemoglobin assay comprises:

converting hemoglobin to a metal-hemoglobin complex; and

detecting the intensity of light responsive to the metal-hemoglobin complex.

3. The method of claim 1, the fluorescent donor dye having an absorption maximum at an excitation wavelength from about 590 to about 660 nanometers.

4. The method of claim 1, the fluorescent acceptor dye having an absorption maximum at an excitation wavelength from about 630 to about 760 nanometers.

5. The method of claim 1, where the fluorescent donor dye is fluorescein isothiocyanate and the fluorescent acceptor dye is tetramethylrhodamine isothiocyanate.

6. The method of claim 1, where the HbA1c analog is any molecule that binds to the HbA1c-specific antibody with less affinity than HbA1c.

7. The method of claim 1, where the HbA1c analog is selected from the group consisting of glycated peptides, multiple antigen peptides, and combinations thereof.

8. The method of claim 1, where the fluorescence of the fluorescent donor dye and the fluorescence of the fluorescent acceptor dye are detected at a wavelength from about 450 to about 520 nanometers or greater than about 600 nanometers.

9. The method of claim 1, the fluorescent acceptor dye replaced with a quenching dye.

10. The method of claim 1, where the change in the intensity includes an increase in the intensity of the donor dye fluorescence and a decrease in the intensity of the acceptor dye fluorescence.

11. The method of claim 1, where the change in the intensity includes a decrease in the intensity of the donor dye fluorescence and an increase in the intensity of the acceptor dye fluorescence.

12. A hemoglobin ratio determination system, comprising:

a vial comprising a lysing mixture;

a sensor strip,

where a total hemoglobin assay is included with the lysing mixture or the sensor strip,

where a HbA1c assay is included with the lysing mixture or the sensor strip, and where the HbA1c assay includes

a fluorescent donor dye attached to a HbA1c analog and a fluorescent acceptor dye attached to an Hb-specific antibody, or

a fluorescent donor dye attached to a Hb-specific antibody and a fluorescent acceptor dye attached to a HbA1c analog, and where

an emission spectrum of the fluorescent donor dye overlaps with an excitation spectrum of the fluorescent acceptor dye; and

an electronic measurement device including a processor, where the processor is operable to determine a ratio of HbA1c to total hemoglobin.

13. The system of claim 12, where the processor determines a comparison of the decrease in the acceptor dye fluorescence in the presence of HbA1c to the acceptor dye fluorescence in the absence of HbA1c.

14. The system of claim 12, where the processor determines a comparison of the increase in donor dye fluorescence in the presence of HbA1c to the donor dye fluorescence in the absence of HbA1c.

15. The system of claim 13, where the processor determines a correlation of HbA1c in the sample responsive to the comparison and the ratio of HbA1c to total hemoglobin is responsive to the correlation.

16. The system of claim 15, where the processor determines the total hemoglobin in the sample substantially before determining the HbA1c in the sample.

17. The system of claim 12, the fluorescent acceptor dye replaced with a quenching dye.

18. A HbA1c sample concentration determination kit, comprising:

a vial including a lysing mixture, a total Hb assay, and a HbA1c assay, the HbA1c assay including one of

a fluorescent donor dye attached to a HbA1c analog and a fluorescent acceptor dye attached to an Hb-specific antibody, and

a fluorescent donor dye attached to a Hb-specific antibody and a fluorescent acceptor dye attached to a HbA1c analog; and

where

an emission spectrum of the donor dye overlaps with an excitation spectrum of the acceptor dye.

19. The kit of claim 18, the fluorescent acceptor dye replaced with a quenching dye.

20. A HbA1c sample concentration determination kit, comprising:

a vial including a lysing mixture and a total Hb assay, and

a sensor strip including a HbA1c assay, the HbA1c assay including one of

a fluorescent donor dye attached to a HbA1c analog and a fluorescent acceptor dye attached to an Hb-specific antibody, and

a fluorescent donor dye attached to a Hb-specific antibody and a fluorescent acceptor dye attached to a HbA1c analog; and

where

an emission spectrum of the donor dye overlaps with an excitation spectrum of the acceptor dye.

21. The kit of claim 20, the fluorescent acceptor dye replaced with a quenching dye.